alpha-synuclein: the gateway to parkinsonism- innsbruck

International Symposium

Alpha-Synuclein

The Gateway to Parkinsonism

11th-13th February 2015

Innsbruck Austria

Host

Innsbruck Medical University

Department of Neurology

Director Werner Poewe

Convenors

Nadia Stefanova

Gregor K Wenning

Venue

Great Lecture Hall MZA Anichstrasse 35 6020 Innsbruck

Austria

Scientific Advisory Board

Glenda Halliday Poul Henning Jensen Leonidas Stefanis Omar El-Agnaf Philipp Kahle Werner Poewe Gregor K Wenning Nadia Stefanova Local Organizing Committee Chairs Daniela Kuzdas-Wood amp Lisa Fellner Edith Sturm Karin Spiss Serena Venezia Dominik Bruumlck Alessandra Fanciulli Florian Krismer Christine Kaindlstorfer Violetta Refolo Gabriele Schobesberger

1

Table of Contents

Course Description and DisclaimerPage 3 Venue and Accreditation InformationhelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellipPage 4 Faculty Listing and Industry Disclosureshelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellip Page 5 Agenda February 11 2015 helliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellipPage 9 Agenda February 12 2015 helliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellipPage 9 Agenda February 13 2015 helliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellip0 Session 5 Abstract Listinghelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellip Page 11

Faculty PresentationshelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellipPage 16

2

Alpha-Synuclein The Gateway to Parkinsonism- Innsbruck Innsbruck Austria | February 11-13 2015 Course Description Course Description

This conference features an internationally renowned faculty of researchers scientists and clinicians who will present on alpha-synuclein related to Parkinsonrsquos disease and parkinsonism during this three-day event

The first dayrsquos sessions will highlight genetics and environment as it is linked to alpha-synuclein advances in PD biomarkers and non-motor aspects of Parkinsonrsquos disease

The second day will address intracellular disruption by alpha-synuclein extracellular alpha-synuclein and multiple system atrophy while the third day will discuss emerging therapies Each session will include a roundtable discussion where participants will be able to interact with the sessionrsquos faculty members and ask pressing questions

In addition participants are invited to submit abstracts for a poster presentation on the second day ndash prize money will be awarded to the top three presentations as judged by the Scientific Advisory Board for the conference

Learning Objectives

At the conclusion of this activity participants should be able to accomplish the following

Define the role of alpha-synuclein in the pathogenesis of PD and MSA Discuss the pros and cons of prion-like propagation in PD and related disorders Identify clinical features in the premotor phase of PD and MSA Critically discuss the value of biomarkers in the early diagnosis of PD and MSA Review disease modification interventions in PD and MSA including immunization

Recommended Audience This course is recommended for neuroscientists and movement disorder neurologists with an interest in alpha-synucleinopathies such as Parkinson`s disease and multiple system atrophy

Evaluations

Please take time to complete the evaluation form provided at this course Your input and comments are essential in planning future educational programs for MDS When completed evaluations may be returned to the registration desk or the MDS International Secretariat Educational Disclaimer The primary purpose of MDS programming is to provide educational opportunities that enhance patient care Information presented as well as publications technologies products andor services

3

Educational Disclaimer continued discussed are intended to inform attendees about the knowledge techniques and experiences of physicians who are willing to share such information with colleagues A diversity of opinions exists in the medical field and the views of the coursersquos faculty are offered solely for educational purposes Faculty membersrsquo views do not represent those of MDS and do not constitute endorsement by MDS MDS disclaims any and all liability for all claims which may result from the use of information publications products andor services discussed at this program

Recordings Prohibited Audio and videotaping are not allowed during the course Photography is also not allowed during the activity

Course Venue

Medizinzentrum Anichstrasse (MZA) Anichstrasse 35 6020 Innsbruck

EACCME Accreditation

The course Alpha-Synuclein The Gateway to Parkinsonism- Innsbruck is accredited by the European Accreditation Council for Continuing Medical Education (EACCME) to provide the following CME activity for medical specialists The EACCME is an institution of the European Union of Medical Specialists (UEMS) wwwuemsnet The course Alpha-Synuclein The Gateway to Parkinsonism- Innsbruck is designated for a maximum of 11 hours of European external CME credits Each medical specialist should claim only those hours of credit that heshe actually spent in the educational activity Through an agreement between the European Union of Medical Specialists and the American Medical Association physicians may convert EACCME credits to an equivalent number of AMA PRA Category 1 Creditstrade Information on the process to convert EACCME credit to AMA credit can be found at wwwama-assnorggointernationalcme Live educational activities occurring outside of Canada recognized by the UEMS-EACCME for ECMEC credits are deemed to be Accredited Group Learning Activities (Section 1) as defined by the Maintenance of Certification Program of The Royal College of Physicians and Surgeons of Canada EACCME credits Each medical specialist should claim only those hours of credit that heshe actually spent in the educational activity The EACCME credit system is based on 1 ECMEC per hour with a maximum of 3 ECMECs for half a day and 6 ECMECs for a full day event Visit the Alpha-Synuclein The Gateway to Parkinsonism- Innsbruck webpage following the course in order to print your course certificate and your EACCME certificate

4

Course Directors

Nadia Stefanova MD PhD Medical University of Innsbruck Innsbruck Austria Nadia Stefanova has no financial relationships to disclose

Gregor Wenning MD PhD Medical University of Innsbruck Innsbruck Austria Gregor Wenning has received a grant from 7FP EU ASF

Course Faculty

Roger Barker BA MBBS MRCP PhD Brain Repair Centre Cambridge United Kingdom Prof Barker receives royalties from Wiley for a Neuroscience book and from Springer for editorial work on JNeurol Paolo Barone MD PhD University of Salerno Napoli Italy Prof Barone has received support as a consultant from Zambon Italia Lundbeck and UCB as well as commercial support for grantsresearch from Lundbeck UCB Eduardo Benarroch MD Mayo Clinic Rochester Minnesota USA Dr Benarroch does not have any commercial relationships to disclose Patrik Brundin MD PhD Van Andel Research Institute Grand Rapids Michigan USA Dr Brundin has received commercial support as a consultant from Capital Technologies Inc Renovo Neural Inc Roche and Teva Lundbeck Additionally he has received commercial support for grantsresearch from Renovo TevaLundbeck Dr Brundin has intellectual property rights in TevaLundbeck and ownership interests in Acousort AB Parkcell AB David Burn MD FRCP Newcastle University Newcastle Upon Tyne United Kingdom Prof Burn has received commercial support for grantsresearch from GSK for study of motilin agonist in PD Additionally he has received royalties from Oxford University Press and Henry Stewart Publishing Paolo Calabresi MD University of Perugia Rome Italy Prof Calabresi has received commercial support for grantsresearch from Bayer Biogen Boehringer Ingelheim Eisai Lundbeck Merck Sharp amp Dohme Novarits and UCB Guumlnther Deuschl MD Christian-Albrechts-University Kiel Germany Prof Deuschl has received commercial support as a consultant from Medtronic Sapiens Britannica and Boston Scientific He has received commercial support for grantsresearch from German Research Council German Ministry of Education and Research and Medtronic Prof Deuschl has received commercial

5

support as honoraria from Medtronic Desitin and UCB He receives commercial royalties from Thieme Publisher Donato Di Monte MD German Center for Neurodegenerative Diseases (DZNE) Bonn Germany Prof Di Monte does not have commercial relationships to disclose Omar El-Agnaf United Arab Emirates University Doha Qatar Dr El-Agnaf does not have any commercial relationships to disclose Karina Fog PhD H Lundbeck AS Copenhagen Denmark Dr Fog has intellectual property rights with and receives salary from H Lundbeck AS Ceacuteline Galvagnion University of Cambridge Cambridge United Kingdom Galvagnion does not have any commercial relationships to disclose Thomas Gasser MD University Department of Neurology Tuumlbingen Germany Prof Gasser has received commercial support as a consultant for Cefalon Pharma and Merck-Serono Additionally he has received commercial support for grantsresearch from Novartis Pharma BMBF Helmholtz Ass European Commission German Research Found and MJFF Prof Gasser has received commercial support as honoraria from Novartis Merck-Serono Schwarz Pharma Boehringer Ingelheim and Teva Pharma He has intellectual property rights from Patent No EP1802749 (A2) KASPP (LRRK2) gene Armin Giese Ludwig-Maximilians-Universitaumlt Muumlnchen Munich Germany Giese has received commercial support as a consultant from BBraun Melsungen and has received commercial support for grantsresearch from MODAG GmbH Giese has intellectual property rights in patents EP1751553 EP2307381WO2010000372 Giese has ownership interests as co-founder and shareholder of MODAG GmbH Glenda Halliday PhD University of New South Wales amp Neuroscience Research Australia Randwick Australia Prof Halliday has received commercial support as honoraria from Novartis Prof Halliday has ownership interests in Cochlear amp NIB Holdings and receives royalties from Academic Press amp Elsevier Halliday receives salary from the commercial interest of University of New South Wales John Hardy MD UCL Institute of Neurology London United Kingdom Prof Hardy has received commercial support as a consultant from Eli Lily Eisai and Cytox Prof Hardy has received commercial support as honoraria from Eisai Takafumi Hasegawa MD PhD Tohoku University Graduate School of Medicine Sendai Japan Prof Hasegawa does not have any commercial relationships to disclose Janice Holton MB ChB PhD FRCPath UCL Institute of Neurology London United Kingdom Dr Holton has received commercial support as honoraria from Lundbeck Henry Houldon MD

6

Dr Houldon does not have any commercial relationships to disclose Poul Henning Jensen MD University of Aarhus Aarhus Denmark Dr Jensen has received commercial support for grantsresearch from H Lundbeck AS Philipp Kahle Univeristy of Tuumlbingen Tuumlbingen Germany Dr Kahle does not have any commercial relationships to disclose Horacio Kaufmann MD The Dysautonomia Research Center New York New York USA Dr Kaufmann has received commercial support as a consultant from Lundbeck Medscape and Astra Zeneca Jeffrey Kordower PhD Rush University Medical Center Chicago Illinois USA Dr Kordower receives salary from the commercial support interest of Rush University Medical Center Seung-Jae Lee PhD Konkuk University Seoul Republic of Korea Dr Lee has received commercial support in the form of honoraria from UCB as well as commercial support for grantsresearch from Dong-A Pharmaceutical Andrew Lees MD Rita Lila Weston Institute of Neurological Studies London United Kingdom Dr Lees has received commercial support in the form of honoraria from Britannia Roche Novartis Boehringer Ingelheim Lundbeck GE Healthcare Servier and Teva Kenneth Marek MD Rita Lila Weston Institute of Neurological Studies London United Kingdom Dr Marek has received commercial support as a consultant from Pfizer GE Healthcare Merck Lilly BMS Piramal Prothena Neurophage nLife Roche and LTI Dr Marek has received commercial support for grantsresearch from GE Healthcare Lilly Piramal Prothena Io Amgen and Bayer He holds ownership interests in Molecular NeuroImaging LLC Eliezer Masliah MD University of California San Diego California USA Eliezer Masliah has received commercial support for grantsresearch from Neuropore Therapies and has other commercial interests in Psychogenics Francisco Pan-Montojo MD PhD University Hospital in Dresden Dresden Germany Dr Pan-Montojo does not have any commercial relationships to disclose

Werner Poewe MD Innsbruck Medical University Innsbruck Austria Prof Poewe has received commercial support as a consultant from AbbVieAllerganAZBIBSciGSKIpsenLundbeckMedronicMSDMerzNovartisTevaUCB and Zambon He has received commercial support as honoraria from AbbVie Allergan AZ BI BSci GSK Ipsen Lundbeck Medronic MSD Merz Novartis Teva UCB and Zambon Prof Poewe receives royalties from Thieme Wiley Blackwell Oxford University Press and Cambridge University Press He holds other commercial interests as consultant and in lecture fees in relation to clinical drug development programs of PD

7

Michael Schlossmacher CM MD Ottawa Hospital Civic Campus Ottawa Canada Dr Schlossmacher has received commercial support as honoraria from Novartis Pharmaceuticals and Teva Neuroscience He has intellectual property rights as the Inventor of patents - licensed to GenzymeSanofi and SNCA ELISA - agreement with Covance Dr Schlossmacher has received royalties from GenzymeSanofi via Brigham amp Womens Hospital and Covance Inc via Ottawa Hospital Achim Schneeberger MD AFFiRis AG Vienna Austria Dr Schneeberger has received commercial support for grantsresearch from Micheal J Fox Foundation and European Union (FP7 program) Dr Schneeberger has received salary from the commercial interest of AFFiRiS AG Klaus Seppi MD Innsbruck Medical University Innsbruck Austria Dr Seppi does not have commercial relationships to disclose Maria Grazia Spillantini FMedSci FRS University of Cambridge Cambridge United Kingdom Dr Spillantini has received commercial support for grantsresearch from MedimmuneAstra Zeneca and Ely Lilli Leonidas Stefanis MD PhD Biomedical Research Foundation Academy of Athens Athens Greece Dr Stefanis has received commercial support as a consultant for AbbVie He has received commercial support as honoraria from Novartis Nadia Stefanova MD PhD Medical University of Innsbruck Innsbruck Austria Dr Stefanova does not have commercial relationships to disclose Eduardo Tolosa MD Hospital Clinic Barcelona Barcelona Spain Dr Tolosa has received commercial support as a consultant from Teva Lundbeck Medtronic UCB and AbbVie He has received commercial support for grantsresearch from MJFox foundationand the Instituto de Salud Carlos III Frederik Vilhardt PhD University of Copenhagen Copenhagen Denmark Dr Vilhardt has received commercial support for grantsresearch from Lundbeck AS Gregor K Wenning MD PhD Medical University of Innsbruck Innsbruck Austria Dr Wenning does not have commercial relationships to disclose

8

Scientific Program

February 11 2015 1200-1210 Opening- Werner Poewe 1210-1235 Keynote Lecture the mysteries of Parkinsonrsquos disease- Andrew Lees 1235-1300 How to define PD- Werner Poewe Session 1 Genetics environment and alpha-synuclein Chairs John Hardy and Thomas Gasser 1300-1315 Clinical genetics overviewndash Thomas Gasser 1315-1330 Limitations of GWAS- Henry Houlden 1330-1345 What is the evidence for environmental factors in PD pathogenesis- Philipp Kahle 1345-1415 Keynote Lecture Genetics and PD- John Hardy 1415-1430 Round Table Discussion 1430-1500 Coffee Break Session 2 Advances in PD biomarkers Chairs Werner Poewe and Michael Schlossmacher 1500-1515 Advances in PD biomarkers CSF- Paolo Calabresi 1515-1530 Advances in diagnostic antibody development and application ndash Omar El-Agnaf 1530-1545 Advances in PD biomarkers peripheral tissues- Jeffery Kordower 1545-1600 Advances in PD biomarkers Imaging- Klaus Seppi 1600-1630 Keynote lecture PPMI- Kenneth Marek 1630-1645 Round Table Discussion 1645-1715 Coffee Break Session 3 Non-motor aspects of PD Chairs Paolo Barone and Eduardo Tolosa 1715-1730 Pre-motor aspects of PD Autonomic failure- Gregor K Wenning 1730-1745 Pre-motor aspects of PD Gastrointestinal disorders- Eduardo Tolosa 1745-1800 Pre-motor aspects of PD Cognitive dysfunction- David Burn 1800-1815 Non-motor sympotoms in advanced PD- Paolo Barone 1815-1830 Neuropathological correlates of non-motor features in PD- Eduardo Benarroch 1830-1900 Keynote lecture Alpha-synuclein propagation underlying motor and non-motor progression

insights from animal models- Patrik Brundin 1900-1915 Round Table Discussion

February 12 2015 Session 4 Intracellular disruption by alpha-synuclein Chairs Philipp Kahle and Maria Grazia Spillantini 0900-0915 Normal function and conformational plasticity of alpha-synuclein- Hilal Lashuel 0915-0930 Autophagy and alpha-synuclein- Leonidas Stefanis 0930-0945 Lysosomes GBA and alpha-synuclein- Michael Schlossmacher 0945-1000 Endoplasmic reticulum stress and alpha-synuclein-

Michael Lee 1000-1015 Synaptic failure and alpha-synuclein- Maria Grazia Spillantini 1015-1030 Alpha-synuclein mediated cellular disruption and lipid packaging- Celine Galvagnion 1030-1100 Round Table Discussion 1100-1400 Session 5 Lunch Break with Poster Tour

9

Complete abstract listing can be found online Session 6 Extracellular alpha-synuclein Chairs Poul Henning Jensen and Glenda Halliday 1400-1415 Mechanisms of unconventional alpha-synuclein secretion-Frederik Vilhardt 1415-1430 How does extracellular alpha-synuclein contribute to pathogenesis and how is it processed-

Seung-Jae Lee 1430-1445 How does exogenous alpha-synuclein get access to the endogenous alpha-synuclein proteins-

Takafumi Hasegawa 1445-1500 Alpha-synuclein progression from enteric neurons- Francisco Pan-Montojo 1500-1515 Viral modeling of alpha-synuclein propagation- Donato Di Monte 1515-1545 Keynote Lecture Neuropathological studies- new insights- Glenda Halliday 1545-1615 Round Table Discussion 1615-1645 Coffee Break Session 7 Multiple system atrophy Chairs Nadia Stefanova and Eliezier Masliah 1645-1700 Clinical presentation and natural history in a North American cohort- Horacio Kaufmann 1700-1715 Neuropathology of MSA an update- Janice Holton 1715-1730 New insights into pathomechanisms- Nadia Stefanova 1730-1745 Preclinical target discovery- Poul Henning Jensen 1745-1815 Round Table Discussion

February 13 2015 Session 8 Emerging therapies Chairs Werner Poewe and Michael Schlossmacher 0900-0915 Whatrsquos in the pipeline for drug therapy- Olivier Rascol 0915-0930 Neurostimulation for PD- Guumlnther Deuschl 0930-0945 Dopamine grafts in PD an update of the TRANSEURO project- Roger Barker 0945-1000 Anle138b ready for clinical trials- Armin Giese 1000-1015 Passive immunization therapies- Karina Fog 1015-1030 Active immunization therapies- Achim Schneeberger 1030-1100 Keynote Lecture What are the bottlenecks for target discovery in synucleinopathies- Eliezer

Masliah 1100-1130 Round Table Discussion 1130-1200 Presentation of Poster Prizes 1200-1215 Closing Remarks

10

Session 5 Abstract Listing ListofPosterPresentations

1

Novel electrochemiluminescence-based ELISA for the detection of a-Synuclein in cerebrospinal fluid of Parkinsonrsquos disease

M G Foslashrland L S Oftedal O-B Tysnes G Alves J P Larsen J Lange Stavanger University Hospital Stavanger Norway

2

The progressive BSSG rat model of Parkinsons disease prodromal indications early asymmetry progressive development α-synuclein aggregates and late stage cognitive deficits

H A Robertson C A Shaw D Baranowski D G Kay J Van Kampen Dalhousie University Halifax Canada

3

Effects of late start MPO inhibition in a preclinical model of Multiple System Atrophy

C Kaindlstorfer P Sommer B Georgievska J Young W Poewe G K Wenning N Stefanova Neurology Innsbruck Austria

4

Synuclein strains show distinct neurotoxic properties in vivo

W Peelaerts L Bousset A Van der Perren C Van den Haute R Melki V Baekelandt KU Leuven Leuven Belgium

5

Non-invasive bioluminescence imagine of alpha-synuclein oligomerization with split firefly luciferase reporters

A Michiels S-A Aelvoet W Werkcx Z Debyser C Van den Haute R Gijsbers V Baekelandt KU Leuven Leuven Belgium

6

Colonic ɑ -synuclein a potential diagnostic biomarker in Parkinsonrsquos disease

C Ruffmann M Hu K Talbot O Ansorge L Parkkinen University of Oxford Oxford United Kingdom

7

Aggregation of monometric human alpha-synuclein (h-α-syn) via crossseeding with environmental proteins in-vitro

C J Wright S E Johnston M Noonan H N Allbutt BMedSci PhD University of Sydney Sydney Australia

8

A time-course study of the response to dopaminergic drugs in mice with 6-OHDA lesions

F Bez V Francardo and M A Cenci Lund University Lund Sweden

9

Salivary alpha-synuclein a new biomarker for Parkinsonrsquos Disease

G Vivacqua A Latorre M Nardi R Mancinelli A Suppa G Fabbrini A Berardelli Department of Neurology Sapienza University of Rome Rome Italy

10 The impact of Rab5A on the formation of glial cytoplasmic inclusion implications for multiple system atrophy

11

D Bruumlck L Fellner G K Wenning N Stefanova Innsbruck Austria

11

The neuroprotective role of AMP-activated protein kinase against the toxicity of intracellular and extracellular α-synuclein in vitro

I Marković M Dulovic M Jovanovic M Xilouri L Stefanis L Harhaji-Trajkovic T Kravic-Stevovic V Trajkovic V Kostic Institute of Medical and Clinical Biochemistry University of Belgrade Serbia

12

Chronic oral administration of rotenone low doses alters gene expression profiles in the gastrointestinal tract of on year-old mice significance for alpha-synuclein aggregation

A Stieacutevenard Y Dening P Bourgois M Schwarz L Catsburg M Meacutequinion T Comptdaer P Semaille O Viltart M-C Chartier-Harlin A Desteacutee F Pan-Montojo C Vanbesien-Mailliot INSERM U837JParc Team 6 Lille France

13

Impairment of Chaperone Mediated Autophagy Induces Dopaminergic Neurogeneration in Rats

M Xilouri O R Brekk A Polissidis L Stefanis Biomedical Research Foundation of the Academy of Athens Athens Greece

14

Activation of autophagy rescues human dopaminergic cells from alpha-synuclein mediated toxicity

N Fussi T Chakroun M Houmlllerhage J Goebel TWRoumlsler G U Houmlglinger DZNE Munich Germany

15

Active immunization against α-synuclein ameliorates degenerative pathology and prevents demyelination in a MSA model

M Mandler E Valera E Rockenstein M Mante H Weninger C Patrick A Adame S Schmidhuber R Santic A Schneeberger W Schmidt F Mattner E Masliah AFFiRiS AG Vienna Austria

16

Evaluation of Anti-α-Synuclein Autoantibody Affinity in Parkinsonrsquos disease and Multiple System Atrophy

T Brudek K Winge K Fog B Pakkenberg L Oslashstergaard Pedersen PhD Research Laboratory of Stereology and Neuroscience Bispebjerg and Frederiksburg Hospital Copenhagen NV Denmark

17

HDAC inhibition protects dopaminergic neurons in a transgenic mouse model of multiple system atrophy

E Sturm P Hockl W Poewe G K Wenning N Stefanova Division of Neurology Department of Neurology Medical University Innsbruck Innsbruck Austria

18

Promoter methylation in the α-synuclein gene is decreased in Parkinsonrsquos disease and correlates with genetic susceptibility variants

L Pihlstroslashm V Berge A Rengmark M Toft University of Oslo Oslo Norway

19

sFIDA a Sensitive Diagnostic Assay for α-Synuclein Aggregates

T Bujnicki S Huumlbinger O Bannach E Birkmann D Willbold Research CenterJuumllich Juumllich Germany

20

Extending the phenotype of transgenic MSA presence of h-ɑ-syn in PNS

D Kuzdas-Wood P Malsch M Theurl N Mair R Irschick J Wanschitz L Klimaschewski W Poewe N Stefanova G K Wenning Medical University Innsbruck Innsbruck Austria

12

21 The role of orexin in Parkinsonrsquos disease A review of article disease

M Salari Isfahan Medical University Isfahan Iran

22

CSF alpha-synuclien species as progression biomarkers for Parkinsonrsquos Disease

N K Majbour K D van Dijk N N Vaikath P Eusebi M T Ardah S Varghese C E Teunissen L BVesterager L P Montezinho H W Berendse L Parnetti W DJ van de Berg and O MA El-Agnaf College of Medical and Health Sciences Al-Ain United Arab Emirates

23

Distinct clinical and neuropathological features of G51D SNC mutation cases compared with SNCA duplication and H50Q mutation

A P Kiely H Ling Y T Asi E Kara P Limousin P Lewis C Proukakis A H Schapira H R Morris S Lubbe N Quinn S Love A J Lees J Hardy T Revesz H Houlden J L Holton UCL Institute of Neurology London United Kingdom

24 High throughput screenings in a neuronal cell model of alpha-synuclein mediated toxicity

M Houmlllerhage M Bickle C Moebius W H Oertel B Hengerer G U Houmlglinger DZNE Munich TU Munich Munich Germany

25

Characterization of the uptake of different α-synuclein forms in primary oligodendroglia compared to an oligodendroglial cell line

L Fellner D Bruumlck G K Wenning N Stefanova MSc Medical University Innsbruck Innsbruck Austria

26

The role of the oligomer(s) in alpha-synuclen aggregation and toxicity impairs the association between SERF protein and RNA

D Otzen N Lorenzen W Paslawski L Giehm C Sahin M Kurnik C Bertelsen J D Kaspersen C M Jessen W Hirst P Reinhart R Staal K Pitts G Krishnamurthy F Mulder J S Pedersen C Betzer P H Jensen iNANO Aarhus University Aarhus C Denmark

27

Alpha-Synuclein beats RNA a competitive amloyid-promoting interaction impairs the association between SERF protein and RNA

H Meyer C Tam-Amersdorfer H Dellago J Grillari S F Falsone University of Graz Graz Austria

28

Olfactory dysfunction in REM-sleep behavior disorder as a model for potential neuroprotection-trials in Lew-body diseases

P Mahlknecht A Iranzo B Houmlgl B Frauscher C Muumlller J Santamariacutea E Tolosa M Serradell T Mitterling V Gschliesser G Goebel F Brugger C Scherfler W Poewe K Seppi Unit of Functional Neurosurgery Sobell Department of Motor Neuroscience and Movement Disorders UCL Institute of Neurology London London United Kingdom

29 Minor salivary gland biopsy for the diagnosis of Parkinsonrsquos disease

T Feng LY Gao Beijing Tiantan Hospital Beijing China

30 Progression Rate of MSA under EGCG Supplementation as anti-Aggregation-Approach

J Levin S Maaszlig M Schuberth A Giese U Mansmann F Krismer GK Wenning K Boumltzel G Houmlglinger Ludwig-Maximilians-Universitaumlt Munich Muumlnchen Germany

31

Alpha-synuclein induces different activation patters in CD4+T in a variant and dose dependent manner which correlates with distinct microglia activation profiles in absence of pathology

J R Christiansen M N Olesen M Romero-Ramos V Sanchez-Guajardo Aarhus University Aarhus Denmark

32 Investigation of intracellular alpha-synuclein multimers in cells and tissue using chemical crosslinking

13

L Reimer J B Lauridsen P H Jensen BA Department of Biomedicine Aarhus C Denmark

33

Identification of synaptosomal proteins binding to monomeric and oligomeric α-synuclein

C Betzer AJ Movius M Shi W-P Gai J Zhang P H Jensen Department of Biomedicine Aarhus University Aarhus Denmark

34

Soluble aggregates of α-synuclein perturb calcium homeostasis by stimulating SERCA

C Betzer J Zheng W-P Gai A Olsen J P Andersen P H Jensen Department of Biomedicine Aarhus University Aarhus Denmark

35

Apnea and dysphagia as the sole features of an α-synucleinopathy

HJ Gilhuis A Panwar SG Van Duinen FJ De Jong MD PhD Erasmus Medical Center Rotterdam Netherlands

36

Elastic abdominal binders attenuate orthostatic hypotension in Parkinsonrsquos disease a randomized controlled trial

A Fanciulli G Goebel BMetzler F Sprenger W Poewe G K Wenning K Seppi Medical University Innsbruck Innsbruck Austria

37

Distinct PET findings separating parkinsonism with alpha-synucleinopathy from tau parkinsonism

C S Lee J Lee J M Koo S Kim S M Kim D Doudet ASAN Medical Center Seoul South Korea

38

In vivo evidence for trans-synaptic degeneration in the striatum of MSA-P patients PET studies with [18F]FP-CIT and [18F]FDG

J M Koo S Kim S M Kim D Kirik C S Lee ASAN Medical Center Seoul South Korea

39

Neuroprotective Role of Regulatory T Cells in Parkinsonrsquos disease Effect of COP-1Alpha-synuclein Vaccination on Pathology Progression

M von Euler Chelpin M Romero-Ramos V Sanchez-Guajardo Aarhus University Aarhus Denmark

40

Early and persistent expression of phosphorylated ɑ-synuclein in the enteric nervous system of A53T mutant human ɑ-synuclein transgenic mice revealed by a week by week study

A Bencsik L Muselli M Leboidre L Lakhdar T Baron Anses Lyons France

41

Alpha-synuclein protofibril-selective antibody prevents motor related death in alpha-synuclein A30P tg mice

F Eriksson V Ramberg J Sigvardson A Kasrayan M Ingelsson P Kahle L Lannfelt G Osswald C Moumlller and E Nordstroumlm BioArctic Neuroscience Stockholm Sweden

42

The molecular tweezer CLR01 reduces alpha-synuclein accumulation and improves motor deficits in a mouse model of Parkinsonrsquos disease

F Richter I Magen P Lee S Subramaniam A Attar J Hayes C Zhu N Franich N Bove K De La Rosa J Kwong G Bitan M- F Chesselet DVM PhD University of Leipzig UMF Institute of Pharmacology Parmacy and Toxicology Leipzig Germany

43 Systematic comparison of the effects of alpha-synuclein mutations on its oligomerization and aggregation

14

D F Lazaro E Rodrigues R Langrohr H Shahpasandzadeh T Ribeiro P Guerreiro E Gerhardt K Kroumlhnert J Klucken M D Pereira B Popova N Kruse B Mollenhauer S O Rizzoli G H Braus K M Danzer T F Outeiro Medical University Goumlttingen Goumlttingen Germany

45

The impact of Myrtus communis essential oil to increase the formation of the toxic aggregate species of alpha-synuclein and enhancement of the cell death

F Aliakbari D Morshedi National Institute of Genetic Engineering and Biotechnology Tehran Iran

44

Alpha-synuclein in minor salivary glands as peripheral biomarkers for sporadic Parkinsonrsquos disease

LY Gao T Feng Beijing Tiantan Hospital Beijing China

45

Screening of small molecules that down-regulate α-synuclein expression

T Asano H Yamakado R Takahashi Department of Neurology Graduate School of Medicine Kyoto University Kyoto Japan

46

Alpha-synuclein methylation in Parkinsonrsquos disease effect of sex and IDOPA

I Schmitt O Kaut L deBoni H Froumlhlich D Berg C Klein H Khazneh U Wuumlllner UKB Bonn Germany

47

A-synuclein oligomers in human red blood cells as a potential biomarker for Parkinsonrsquos disease

F Li P Liu X Li L Gao T Feng Beijing Tiantan Hospital Beijing China

48

Intracellular processing of disease-associated ɑ-synuclein in the human brain suggests prion-like cell-to-cell spread

G G Kovacs L Breydo R Green V Kis G Puska P Lőrincz L Perju-Dumbrava R Giera W Pirker M Lutz I Lachmann H Budka V N Uversky K Molnaacuter L Laacuteszloacute Medical University Vienna Vienna Austria

49

Modeling early stages of Parkinsonrsquos disease in vivo are we there yet

C Vanbesien-Mailliot A Stieacutevenard O Viltart Universiteacute de Lille Lille France

15

Faculty Presentations

16

Advances in PD biomarkers peripheral tissues

Jeffrey Kordower PhD Rush University Medical Center Chicago Illinois USA

252015

1

Jeffrey H Kordower PhD

Department of Neurological SciencesRush University Medical Center

Sensitive

Discriminative Simple

Inexpensive

Example of a good biomarker

blood tests CSF test but they are not discriminative for PD

Control 1 years PD 4 years PD

5 years PD 11 years PD 15 years PD 21 years PD

TH immunohistochemistry in the Putamen of human brain

3

252015

2

Premotor symptoms olfactory dysfunction autonomic dysfunction constipation depression anxiety REM behavior disorder executive dysfunction

Signsfindings presynaptic darr DA uarrechogenicity of the SN hyposmia slight motor signs

Berg Neurodegenerative Diseases 20085133

50 darrDAneurons atsymptom onset rapiddecline over 5y

Parkinsonrsquos disease is caused by exposure to

lipopolysaccharide (bacterial endotoxin) which promotes α-synuclein

overexpressionaggregation in neurons of the

submucosal plexus of the gastrointestinal system then spreads via neural networks to

the medulla then rostally with cumulative

neurological signs

medulla oblongata

pontine tegmentum

basal mid and forebrain

amygdala hypothalamus thalamus

cerebral cortex mesocortex

cerebral cortex neocortex

SN IV

DMN X II raphe II LC II

SN IV

Aurbach V

Braak amp Del Tredici Neurology 2008701916

252015

3


Braak et al Neuroscience Lettters 200639667-72

ldquohellipa putative environmental pathogenhellipmight

induce α-synuclein misfolding and aggregationin specific cell types of the submucosal plexus and reach

the brain via a consecutive series of projection neuronsrdquo

Grafts of dopamine cells placed into the striatum with viral over-expression of alpha synuclein

Note the physical segregation of the graft (brown) and gene delivery (black)

Kordower et al Neurobiology of Disease in press

252015

4

A small percentage (5) of grafted neurons retrogradely transported host-derived alpha synuclein


Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns

Kordower et al Nat Med 200934254

1965-1968Honolulu Heart Program

8006 Japanese

1991-1993Honolulu-Asia Aging Study

3741 Japanese (80 of survivors of HHP)

252015

5

Incidence10K person-years

Bowel

movements

Sample size Incident PD

cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39

Test for trend P=0002 P=0005

Overall 6790 96 75

Honolulu Health amp Aging Study Abbott et al 200157(3)456

bullAppears highest in men with lt1BMday and laxatives gt 2week 51610000 person years

Can we demonstrate early GI involvement in PD biopsy distal colon in early PD

Is the gut ldquoleakyrdquo in PD subjects intestinal permeability studies

evidence for endotoxin exposure Systemic

local

Do PD subjects have ldquodysbiosis (alterations in intestinal flora)rdquo DNA ldquofingerprintingrdquo

Early untreated ldquoclassical PDrdquo no coagulopathy alcoholism occupational

exposure to microbes primary GI disease

Unprepped flexible sigmoidoscopy with biopsy immunohistochemistry (synuclein nitrotyrosine)

Differential sugar absorption sugar ingestion

24-hour urine collection with assays

16S rRNA ldquofingerprintingrdquo for dysbiosis

252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median

Controls (N=26) Crohnrsquos (N=14) Ulcerative colitis (N=11)

Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7

Alpha synuclein Nitro-tyrosine

A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis

Aged-Matched Control

A

C

B

D

α-synuclein N-tyrosine 85-yo woman

Psychotic depression 2002rarrECT

MCI

Rest tremor 22010

Colonic polyp biopsied 2005

MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric

Lactosemannitol Enteric

Sucralose colonic

24-hour sucralose excretion

0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016

1 kg bacteria most unknown identity most anaerobic luminal population amp mucosal biofilm

1012 microbesml luminal content Established by age 2 stable Important becausehellip

genetic component (100x as many geneshellip) metabolic efficiency energy homeostasis important in systemic inflammation HIV coronary artery disease insulin resistance EtOH liver diseasehellip

Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces

Gram (+) coccobacilli

methanobrevibacter

Salminen et al Br J Nutr 199880S147

252015

10

Method Advantages Disadvantages

morphologybiochemistry straightforward cheap subjective culturable only

specific biomarkers may not require culture canrsquot help with unknown

species requires unique biomarker

ribotyping (RNA

polymorphisms)

reliable high

discriminatory power

culturable only

16S ribosomal RNA typing high fidelity reliable

cumulative database culturable amp non-

culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)

Gender male 70 55 31 46

Duration PD (y)

Median (range)

15 (05-8)

Total UPDRS 22 (15-28)

HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11

PCO case scores (Bray Curtis)

Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)

tUPDRS HY BMd α-synuclein nitrotyrosine Abn mucosa Abn

lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++

74M 4 28 2 lt1 ++ ++ radic radic

68F 05 24 2 2 ++++ radic radic44F 2 16 1 lt1 ++++ radic48M 8 18 2 1 ++++ ++++

60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13

GI changes such as a leaky gut or alpha synuclein in thecolon demonstrates excellent sensitivity

Colonic biopsies can be a sensitive and inexspensive biomarker

Still to be proven is how specific this potential biomnarker is and studies in PSP and MSA patients are currently underway

Kathleen Shannon MDAli ]Keshavarzian MD

Christopher Forsyth

Hemraj DodiyaLeo Kelly

Pre-motor aspects of PD Autonomic failure

Gregor K Wenning MD PhD Medical University of Innsbruck Innsbruck Austria

252015

1

Premotor PD autonomic failure Prof Gregor K Wenning

Premotor PD autonomic failure

Prof Gregor K Wenning

Division of Neurobiology


Medical University of Innsbruck


Non-motor symptoms in PD

bull Autonomicbull OH constipation impotence urinary incontinence or retention hyperhidrosis

bull Hyposmia

bull Sleep

bull Cognitive deficitsbull Executive dysfunction MCI dementia

bull Psychiatric deficitsbull Depression psychosis anxiety apathy

bull Otherbull pain


Lim amp Lang 2009

NMS in PDExtranigral pathology

252015

2


ILBD PANS InvolvementBloch et al 2006


2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc

Autonomic failure in premotor PD


Premotor PD Cardiovascular dysfunction

1 Orthostatic hypotension (OH)

2 Chronotropic insufficiency

3 Heart rate variability

4 Cardiac sympathetic imaging

Palma amp Kaufmann 2014

252015

3


Orthostatic hypotension- Consensus criteria (Freeman et al 2011) -

1 Abnormal blood pressure drop (∆ 2010)within 3 minutes of standing or tilt

2 Abnormal blood pressure drop (∆ 2010)after 3 minutes of standing or tilt = delayed OH


OH SymptomsCerebral hypoperfusion

bull Light-headedness

bull Dizziness

bull Visual disturbances

bull Loss of consciousness

bull Impaired cognition

Muscle hypoperfusion

bull Coat-hanger ache

bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue

Modified from Mathias 1999


2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)

Early premotor PD-OH

252015

4


Prodromal autonomic features in PD amp DLBPostuma et al 2013

Systolic drop erectile

dysfunction and

constipation could

identify disease up to 5

years before diagnosis

with sensitivity ranging

from 50 to 90


bull Prospective cohort studybull 14-year follow-up

bull gt 5000 adults aged ge 65 yearsbull Premotor OH in 18 of 214 incident PD cases

bull OH in 18 of non-PD controls

No evidence of premotor OH

MDJ 2012

Premotor OH


Chronotropic insufficiencyMDJ 2013MDJ 2013

Retrospective cohort study2539 patients without neurological diseaseCardiac stress testing at baseline18 incident PD cases within 4 yearsBlunted heart rate compared to non-PD cohort

Chronotropic insufficiency a sign of cardiacsympathetic denervation

Chronotropic insufficiency

252015

5


Heart rate variability

1 HRV parameters are reduced in PD and iRBD (Valappil 2010 Palma

2013)

2 BUT Prospective studies failed to prove a predictive value (Postuma

2010 Jain 2012 Palma 2013)

number of incident PD too small only linear HRV parameters used


Cardiac sympathetic denervation- Kashihara 2006 Orimo 2008 -

MDJ 2012PDHCMSA


Cardiac sympathetic denervationidentifies premotor PD

MDJ 2013

252015

6


Constipation a strong PD risk factor


Premotor sexual dysfunction


50 patients 66 yrs DD 2 yrs HY 19Urinary storage symptoms 64 voiding symptoms 28Urodynamics detrusor overactivity 58 underactivity 50Few patients with urge incontinenceNo sphincter EMG abnormalities

No correlation with gender disease severity or motor type

Urinary biomarkers

252015

7


Feature MSA PAF PD

Site Preganglionic Postganglionic Postganglionic

Prognosis Poor Survival gt 10-20 yrs Prolonged survival

Motor features Always Rare Always

Neuropath Marker GCI LBs LBs

OH Common Common Less common

Sleep apnea Common Uncommon Common

Urinary dysfunction Early Late Intermediate

Supine NE Normal Very low Intermediate

Orthostatic NE rise Subnormal Subnormal Intermediate

Postsynapticadrenoreceptor

supersensitivity

Mildly increased Increased Mildly increased

Garland et al 2013

PAF among the primary autonomic failure syndromes


Conclusions

bull Autonomic failure is an early feature of PD

bull No single autonomic feature accurately predicts PD

bull Low specificity of some autonomic features

bull Identification of autonomic features to be improved

bull No premotor data for some autonomic features such as sweating disorder


Division of NeurobiologyDepartment of Neurology

252015

8


Acknowledgements

Pre-motor aspects of PD Cognitive dysfunction

David Burn MD FRCP Newcastle University Newcastle United Kingdom

Pre-Motor Aspects of PD Cognitive Dysfunction

David J Burn davidburnnclacuk

Introduction

bull Little or no direct evidence

bull Strong circumstantial data bull Implausible to think

cognitive deficits do manifest in premotor phase

Review of Evidence

Postuma 2012

DLB versus PDD

Halliday amp McCann 2009

Lumping versus Splitting

bull DLB introduced 20 years ago to differentiate LB dementia from AD ndash lack of focus on cognition in PD at time

bull PDD and DLB are VERY similar ndash clinical syndrome neuropathology neurobiology

bull Problem ndash confusing terminology ndash one-year rule difficult to justify ndash complicates advocacy amp research

Redefining PD

MCI at Disease Onset I

0

5

10

15

20

25

MCI at Disease Onset II

Rate of Cognitive Decline

Maetzler 2009

Braak amp Cortical Pathology

Braak 2003

These findings broadly support the Braak hypothesis of caudo-rostral development but indicate that the extent of the disease in the cortex and the severity of pathology in the medulla were independent of one

another

DaT Binding amp Outcome

Ravina 2012

Estimated differences in continuous outcomes by quartile of baseline mean striatal binding

ORs for dichotomous outcomes by quartiles of baseline mean striatal binding

Early MRI Markers amp Cognition

Duncan under review

Early CSF Changes amp Cognition

Hall 2015


Stewart 2014

Cognition amp GBA Mutations

Zokaei 2014


Zokaei 2014

Double dissociation between sources of error in VSTM associated with GBA mutation amp PD

iRBD amp Cognitive Deficits

Gagnon 2009

MCI Subtypes iRBDPatients

PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)

Amnestic MCI single domain n () on totalnumber of MCI cases

2 (125) 0 0 0

Nonamnestic MCI single domain

Executive functionsattention n 9 7 1 3

Visuoconstructionalvisuoperceptual abilities n 0 0 1 0

Total n () on total number of MCI cases 9 (56) 7 (44) 2 (100) 3 (100)

Amnestic MCI multiple domain

Verbal learningmemory and executivefunctionsattention n

1 4 0 0

Verbal learningmemory andvisuoconstructionalvisuoperceptual abilities n

0 0 0 0

All domains n 2 2 0 0

Total n () on total number of MCI cases 3 (19) 6 (37) 0 0

Nonamnestic MCI multiple domain n () ontotal number of MCI cases

2 (125) 3 (19) 0 0

Patients with idiopathic RBD have up to an 80 risk of developing a neurodegenerative synucleinopathy (Postuma 2014)

TCS amp Premotor Cognition

Berg 2011

Conclusion

bull No direct evidence for pre-motor cognitive dysfunction in PD

bull But indirect data persuasive amp suggests may be common if mild

bull What does this tell us about pathophysiology

Neuropathological correlates of non-motor features in PD

Eduardo Benarroch MD Mayo Clinic Rochester Minnesota USA


Eduardo E Benarroch MD

Parkinson disease (PD) is a neurodegenerative disorder affecting essentially all levels of the central and peripheral nervous system and characterized both by motor and non-motor features The typical motor features of PD reflect accumulation of α-synuclein (α-Syn) containing Lewy bodies (LB) and Lewy neurites (LN) associated with loss of dopaminergic neurons of the substantia nigra pars compacta (SNc) However there is clear evidence that PD manifests with non-motor symptoms that reflect involvement of other levels of the neuraxis Some of these non-motor symptoms may antecede the development of parkinsonism for many years (ldquopremotorrdquo symptoms) whereas others occur ate later stages of disease Non-motor symptoms include autonomic sleep behavioral cognitive and sensory manifestations The evolution of these symptoms tends in general to follow the neuropathological stages of LBLN neuropathology described by Braak et al

Autonomic manifestations The most common autonomic manifestations of PD are constipation and urogenital dysfunction others include orthostatic hypotension (OH) and sudomotor abnormalities Constipation typically precedes the motor symptoms and reflects involvement of the enteric nervous system (ENS) at the earliest stages of disease There is also involvement of the dorsal motor nucleus of the vagus (DMV) at Braakrsquos stage I The DMV primarily controls upper gastrointestinal function and its involvement may manifests with esophageal and gastric dysmotility rather than constipation Drooling reflects impaired swallowing as PD is associated with reduced salivary secretion reflecting the early involvement of the salivary glands by α-Syn neuropathology Dysphagia defecatory dysfunction and bladder symptoms in PD are multifactorial and reflect not only involvement of autonomic effectors but also central pattern generators For example dysphagia may reflect afferent or efferent denervation of the pharynx but so far there is no evidence of neuronal loss in the nucleus ambiguus However accumulation of LBLN in the medullary reticular formation containing the central pattern generator for swallowing occurs at relatively early stages of disease Similarly defecatory dyssynergia and detrusor hyperactivity may reflect involvement of ldquogain settingrdquo structures in the brainstem in addition to LBLN accumulation in the sacral parasympathetic nucleus (innervating the rectum and bladder) and the Onuf nucleus (innervating the external sphincters and pelvic floor Unlike the case of multiple system atrophy (MSA) in general OH is either a mild or late manifestation of PD except in approximately 20 of cases OH likely reflects involvement of peripheral and central components of the sympathetic cardiovascular control There early sympathetic denervation of the heart and accumulation of LBsLNs in sympathetic ganglia intermediolateral cell column (IML) and rostral ventrolateral medulla (RVLM) which provides tonic sympathoexcitatory input to the IML controlling blood pressure Whereas LBsLNs accumulate in the RVLM at early stages of disease neuronal loss in this area is typically less consistent and severe than in MSA Skin biopsy studies show accumulation of α-Syn in axons innervating the epidermis sweat glands and pilomotor nerves The clinical correlations are still undetermined but they may contribute to sudomotor and pain manifestations of PD

Sleep disorders The various manifestations of sleep dysregulation in PD likely reflect involvement of brainstem and hypothalamic systems involved in arousal and sleep sleep-state switch These

include cholinergic neurons of the basal forebrain and pedunculopontine tegmental nucleus (PPTPPN) noradrenergic neurons of the locus ceruleus (LC) serotonergic neurons of the caudal and rostral raphe dopaminergic neurons of the ventral periaqueductal gray and orexin neurons of the lateral hypothalamus All these regions show LBsLNs as well as neuronal loss at intermediate

stages of PD Whereas involvement of these regions likely contributes to excessive diurnal somnolence in PD the anatomical substrate of REM sleep behavior disorder (RBD) which is on the earliest non-motor manifestations of PD is still undetermined Experimental and lesional studies suggest that RBD may reflect involvement of the sublaterodorsalsubceruleus region of the pons containing REM-on neurons andor the nucleus gigantocellularis of the medulla which receives inputs from these neurons and mediates inhibition of motoneurons during REM sleep Involvement of the ldquogain-controlrdquo cholinergic and monoaminergic groups mentioned above may be contributory although the relationship between neuronal loss in these areas and RBD has been inconsistent

BehavioralPsychiatric Manifestations The anatomical bases of behavioral and psychiatric manifestations of PD including depression and anxiety are still speculative Abundant experimental and pharmacological evidence indicates that these manifestations may at least in part reflect involvement of the rostral raphe and locus ceruleus as well their targets including the amygdala as well as the orbitofrontal and anterior cingulate cortices Involvement of dopaminergic neurons in the ventral tegmental area innervating these cortical regions may contribute to abulia

Dementia Dementia in PD affects predominantly the executive and visuospatial domains There is evidence that the main substrate of dementia in PD is cortical accumulation of LBsLNs For example development of hallucinations reflects LBLN accumulation in the lateral temporal cortex A substantial proportion of patients with PD and dementia also have Alzheimer-type neuropathology including accumulation of amyloid plaques and tau-containing neurofibrillary tangles in the cortex These changes contribute to the severity of dementia in these patients

Sensory Symptoms PD is associated with disturbances in olfactory visual and nociceptive processing Olfactory dysfunction is an early finding in PD and reflects early involvement of the olfactory bulb by α-Syn neuropathology visual disturbances may reflect retinal involvement (eg amacrine cells) However at late stages of disease olfactory and particularly visual symptoms (including hallucinations) likely reflect involvement of the olfactory and visual association areas Pain is a common manifestation of PD and is multifactorial Involvement of cutaneus nerves and superficial laminae of the dorsal horn may be contributory in some cases

In summary There are multiple potential substrates for both the early or late non-motor manifestations of PD Whereas there is accumulation of LBsLNs in all the areas mentioned in this summary the relationship between α-SYN accumulation and clinical symptoms remains for the most part speculative Whereas pathological α-Syn probably disrupts axonal transport vesicular dynamics and thereby cell function and survival the relationship between LBLN accumulation and selective neuronal vulnerability leading to cell loss are still incompletely understood For example whereas exposure to environmental toxins may explain early ENS involvement monoaminergic phenotype the early LC or raphe involvement and the presence of long small myelinatedunmyelinated axons the DMV involvement these phenotypic characteristics are not necessarily shared by all neuronal groups affected by the disease Nevertheless these findings emphasize the need to interpret PD as a systemic neurologic disorder

Keynote lecture Alpha-synuclein propagation underlying motor and non-motor progression insights from animal models

Patrik Brundin MD PhD Van Andel Research Institute Grand Rapids Michigan USA

Alpha-synuclein propagation underlying motor and non-motor progression

insights from animal models

Patrik Brundin MD PhD

Laboratory of Translational Parkinsonrsquos Disease Research

Center for Neurodegenerative Science Van Andel Research Institute Grand Rapids MI USA

patrikbrundinvaiorg

In 2008 observations of Lewy pathology in post-mortem studies of dopamine neurons grafted to the brains of Parkinsonrsquos disease (PD) patients kindled the idea that PD is a prion-like disorder The alpha-synuclein (a-syn) aggregates found in the transplanted neurons displayed several features that are characteristic of Lewy pathology eg they were ubiquitinated Thioflavin S-positive and stained for the P129S post-translational modification A review of all post-mortem studies of transplanted PD patients suggests that a-syn aggregates develop in transplants following a lag time of one decade after surgery The observations made in transplanted neurons might also explain why Lewy pathology develops progressively following a stereotypic pattern consistent with neural connections (so called Braak stages) in the non-grafted PD brain While Braak and coworkers proposed that a neurotropic virus explains why a-syn aggregates seem to first develop in anterior olfactory structures and the dorsal motor nucleus of the vagal nerve in PD reaching the substantia nigra dopaminergic neurons only several years later the emerging evidence now suggests that misfolded a-syn itself is the ldquospreading agentrdquo Thus misfolded a-syn behaves in a prion-like fashion This gradual propagation of a-syn pathology throughout the nervous system might also in part explain the progression of symptoms in PD

Since 2008 numerous experiments in cell cultures and animal models have clearly demonstrated that a-syn transfers between cells The research field has also begun to address the molecular mechanisms that underpin spreading of a-syn aggregates from one cell to another Other experiments have focused on modeling how a-syn neuropathology propagates over long distances in PD along axonal tracts from one brain region to another In this presentation I will describe factors affecting the release of a-syn from neurons and the uptake of a-syn in neurons oligodendrocytes and microglia I will discuss how a-syn is handled once it has entered a new cell I will also describe how the intercellular transfer of a-syn can be modeled in experimental animals In the future a better understanding of the prion-like behavior of a-syn can lead to the identification of novel molecular targets for therapies that slow the progression of PD

Alpha-synuclein mediated cellular disruption and lipid packaging

Celine Galvagnion University of Cambridge Cambridge United Kingdom

Lipid vesicles trigger α-syn aggregation by stimulating primary nucleation

Dr Ceacuteline Galvagnion

12022015

Alpha-Synuclein The Gateway to Parkinsonism ndash Innsbruck 2015

Galvagnion C et al Nat Chem Biol 2015

α ndash synuclein and lipids

bull  α-syn is present in the cells as two distinct forms a cytosolic natively disordered conformation and an α-helical membrane-bound form

bull  In addition the interaction between α-syn and lipids has been shown either to enhance or to suppress amyloid fibril formation

bull  The binding of α-syn to membrane vesicles affects the properties of the membrane

Dr C Galvagnion 12 02 15

Fusco G et al Nat Commun 2014

Ouberai MM et al J Biol Chem 2013

Auluck PK et al Annu Rev Cell Dev Biol 2010

Our approach

bull  Main Goal bull  Understand how membrane vesicles modulate the kinetics of amyloid formation of α-

syn bull  Establish a detailed and mechanistic description of the role that lipid vesicles play on α-syn aggregation

bull  Lipid system synthetic vesicles made of phosphatidylserine lipids whose sizes range from 20 to 100 nm


α-syn binding to lipid vesicles

bull  α-syn folds as an α-helix upon binding to vesicles

-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b

eacute [lipid vesicles]



bull  α-syn binds to vesicles with a sub micro molar affinity and 30 lipid molecules are involved in the binding of one monomeric molecule of α-syn

-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60

KD = 38 plusmn 13 10-7 ML = 282 plusmn 08

b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate


Modulation of α-syn aggregation by lipid vesicles

a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e

bull  For high [α-syn][vesicle] ratios the presence of vesicles strongly enhances the rate of aggregation of α-syn

bull  No amyloid formation was observed when α-syn was incubated bull  i) in the absence of vesicles bull  ii) in the presence of a large excess of vesicles

Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)

No vesicle or excess of vesicles

High [α-syn][vesicle] ratios



b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20

20 μM α-syn seeds [DMPS](mM)

842

bull  When pre-formed fibrils were incubated with an excess of vesicles the protein was found to dissociate from the fibrils and populate the lipid-bound monomeric α-helical state

bull  The lipid-bound a-helical state of α-syn is thermodynamically more stable than the fibrillar state under these conditions


α-syn and vesicles have distinct role in amyloid formation

bull  The concentration of protein molecules converted into fibrils was found to be

bull  constant for all the different initial concentrations of α-syn free in solution

bull  proportional to the initial concentration of α-syn bound to the vesicles

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3

[α-synbound] = 10 μM

[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)

[α-synfree] = 140 μM

2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d



Oslash  The rates of secondary nucleation and fragmentation are negligible

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d


Morphologies of the fibrils formed by α-syn in the presence of vesicles

bull  Two distinct types of structures are present

-  small spherical species with a diameter of 50 nm agrave vesicles coated with α-syn

-  thin filaments that appear to be attached to the vesicles agrave amyloid fibrils

bull  The average length of the fibrils observed indicates that on average only a single nucleation event had occurred at each vesicle


Why does the reaction of amyloid formation end when free monomers are still present in solution

Is the remaining free monomer inactive Are the fibril ends no longer growth competent



Monomeric α-syn is still capable of nucleating at the surface of lipid vesicles

Oslash  The reaction of amyloid formation ceases because of the depletion of nucleation sites on the vesicles and the inactivation of growing fibril ends and not because of the depletion of free monomeric α-syn

Monomeric α-syn is still capable of elongating fibrils


Analysis of the kinetics of amyloid formation

Knowles TP et al Science 2009 Cohen SI et al J Chem Phys 2011



Model -  Heterogeneous primary nucleation (Campioni S et al JACS 2014 Giehm L et al

Methods 2011)

-  No fragmentation nor secondary nucleation


Vesicles can enhance the nucleation rate by a thousand fold

The rate of primary nucleation is enhanced by at least three orders of magnitude by the presence of vesicles


Conclusions

bull  Taken together our results provide a self-consistent explanation for the observed modulation of α-syn amyloid formation by lipid vesicles

bull  At low α-synvesicle ratios all the protein molecules are bound to the surface of the vesicles in a predominantly helical conformation no fibril formation is observed

bull  At high α-synvesicle ratios the rate of heterogeneous primary nucleation of α-syn can be enhanced by at least three orders of magnitude relative to that occurring in bulk solution and at the airwater interface

low α-synvesicle ratios high α-synvesicle ratios


Acknowledgements

Thomas CT Michaels

(TPJKrsquos group) Prof Christopher M

Dobson

Dr Alexander K Buell

Dr Tuomas PJ Knowles

Prof Michele Vendruscolo

Georg Meisl (TPJKrsquos group)


How does extracellular alpha-synuclein play roles in pathogenesis and how is it processed

Seung-Jae Lee PhD Konkuk University Seoul Republic of Korea

How does extracellular alpha-synuclein play roles in the pathogenesis and how is it processed

Seung-Jae Lee

Department of Biomedical Science and Technology Konkuk University Seoul Korea

Genetic studies have implicated protein aggregation and lysosomal dysfunction in the pathogenesis of Parkinsonrsquos disease (PD) Deposition of α-synuclein aggregates occurs widely in the central and peripheral nervous systems in PD Although recent evidence has suggested that cell-to-cell transmission of α-synuclein aggregates drives the progression of PD the mechanism by which α-synuclein aggregates spread remains undefined Here I present the evidence that α-synuclein aggregates are perpetually transmitted through a continuous cycle involving uptake of external aggregates co-aggregation with endogenous α-synuclein and exocytosis of the co-aggregates Moreover we found that glucocerebrosidase 1 depletion which has previously been strongly associated with PD and increased cognitive impairment promoted propagation of α-synuclein aggregates Depletion of other genes such as ctsd (cathepsin D) and ATP13A2 resulted in mixed outcomes in lysosomal functions The cell lines with these gene depletions further confirmed that lysosomal dysfunction is the key modulator of spreading of synucleinopathy These studies define how α-synuclein aggregates spread among neuronal cells and explain how lysosomal dysfunction increase the risk of developing PD and other synucleinopathies

How does exogenous alpha-synuclein get access to the endogenous alpha-synuclein proteins

Takafumi Hasegawa MD Tohoku University Sendai Japan

Viral modeling of alpha-synuclein propagation

Donato Di Monte MD German Center for Neurodegenerative Diseases (DZNE) Bonn Germany

252015

1

Donato A Di Monte MD

ldquoV IRAL MODELING OF αααα-SYNUCLEIN PROPAGATIONrdquo

AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE

BETTINA WINZEN-REICHERT

FRANZISKA HESSE

ACKNOWLEDGEMENTS

MODELING αααα-SYNUCLEIN INTER-NEURONAL PROPAGATION

SITE OF INITIAL PATHOLOGYCAN INCREASED αααα-SYNUCLEIN EXPRESSION BE INDUCED IN THE MEDULLA OBLONGATA OF EXPERIMENTAL ANIMALS

TRIGGERING MECHANISMCAN αααα-SYNUCLEIN PROPAGATION BE TRIGGERED BY SIMPLY ENHANCING ITS INTRANEURONAL LEVELS

ααααENHANCED αααα-SYNUCLEIN EXPRESSION IS ITSELF

CAPABLE OF CAUSING HUMAN PARKINSONISM

RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS

FARRER M ET AL (2004) ANN NEUROL 55174-

179

BRAAK H ET AL (2003) NEUROBIOL AGING 24197-211MEDULLAOBLONGATA

ααααIN TYPICAL PD αααα-SYNUCLEIN PATHOLOGY BEGINS IN

THE LOWER BRAINSTEM AND SPREADS FOLLOWING A

CAUDO-ROSTRAL PATTERN

PATHOGENETIC MECHANISMS


INTRAPARENCHYMAL (EG INTRANIGRAL ) INJECTION OF VIRAL VECTORS (EG AAV) CARRYING THE

αααα-SYNUCLEIN GENE HAS LONG BEEN USED AS AN EXPERIMENTAL STRATEG Y TO INDUCE αααα-SYNUCLEIN

OVEREXPRESSION AND CONSEQUENT PATHOLOGY

EXPERIMENTAL MODELS

SIDE INJECTED WITH AAV- αααα-SYNINTACT SIDE

TH

LOSS OFTH-POSITIVE NEURONS (ARROW) IN THE SUBSTANTIA NIGRA OF A MOUSE UNILATERALLY

INJECTED WITH A VIRAL VECTOR CARRYING THE αααα-SYNUCLEIN GENE (AAV-αααα-SYN)

CAN VIRAL VECTORS BE USED TO INCREASEαααα-SYNUCLEIN

PRODUCTION IN THE MEDULLA OBLONGATA OF EXPERIMENTAL

ANIMALS (EG RATS)

IF SO WOULD THIS OVEREXPRESSION BE FOLLOWED BY

CAUDO-ROSTRALαααα-SYNUCLEIN PROPAGATION

252015

2


VIRAL VECTORS CARRYING THEDNA FOR HUMAN αααα-SYNUCLEIN ARE

INJECTED INTO THE VAGUS NERVE IN

THE RAT NECK

1

αααα-SYNUCLEIN IS OVER-EXPRESSED WITHIN

NEURONS IN THE MEDULLA OBLONGATA

VECTORS TRAVEL VIA THE NERVE X TO

REACH THE LOWER BRAINSTEM2

3

3

LOWER BRAIN

4

1

VAGUS NERVE

2

THE RAT VAGUS NERVE ISCOMPRISED OF AXONS THATORIGINATE FROM ORTERMINATE IN THE MO ANDUPPER CERVICAL SPINAL CORD

THE PRESENCE OF EXOGENOUS(HUMAN) αααα-SYNUCLEIN IN REGIONS ROSTRAL TO

THE MEDULLA OBLONGATA (EG PONS) WOULD INDICATE NEURON-TO-NEURON

PROTEIN TRANSMISSION

4

BECAUSE OF THE ANATOMICAL CONNECTIONS OF FIBERS FORMING THE V AGUS NERVE SUCCESSFULAND SPECIFICAAV- INDUCED

TRANSDUCTION WOULD BE INDICATED BY OVEREXPRESSION OF HUMAN αααα-SYNUCLEIN WITHIN

(1) CELL BODIES AND AXONS IN THE DORSAL MOTOR NUCLEUS OF THE VAGUS NERVE IPSILATERAL TO AAV INJECTIONS

(2) AXONAL PROJECTIONS IN THE SOLITARY TRACT IPSILATERALAND CONTRALATERAL TO AAV INJECTIONS AND

(3) CELL BODIES AND AXONS IN THE NUCLEUS AMBIGUUS IPSILATERAL TO AAV INJECTIONS


RESULTSBREGMA (mm) -1428 -1404 -1376 -1296 -1200

IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN

NEURONAL CELL BODIES AND AXONAL PROJECTIONS IMMUNOREACTIVEFOR HUMAN αααα-SYNUCLEIN IN THE DORSALMOTOR NUCLEUS OF THE VAGUS NERVE(DMV) AND NUCLEUS AMBIGUUS(NA) IPSILATERAL TO AAV INJECTIONS

DMV NA

CAUDO-ROSTRAL PROTEIN SPREADING WOULD BE INDICATED BY THE PRESENC E OF EXOGENOUS (HUMAN ) αααα-SYNUCLEIN IN AREAS

PROGRESSIVELY DISTANT FROM THE MEDULLAOBLONGATA (SITE OF OVEREXPRESSION)


RESULTS

18 W

EE

KS

RMB

FB

FB = FOREBRAINRMB = ROSTRAL MIDBRAIN

8 W

EE

KS

PONS

CMB

CMB = CAUDAL MIDBRAIN

4 W

EE

KS

MO

MO = MEDULLA OBLONGATA

PREDILECTION

SITES

COERULEUS-SUBCOERULEUS COMPLEX

HYPOTHALAMUS AMYGDALA

DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN

AT 4 WEEKS POSTAAV INJECTIONS HUMAN αααα-SYNUCLEIN WAS STILL CONFINED WITHINTHE MEDULLA OBLONGATA NO EVIDENCE OF PROPAGATION

AT LATER TIME POINTS (8 AND 16 WEEKS) AXONAL PROJECTIONSIMMUNOREACTIVE FOR HUMAN αααα-SYNUCLEIN WERE FOUND IN BRAINREGIONS PROGRESSIVELY ROSTRAL TO THE MEDULLA OBLONGATA

SPREADING FOLLOWED A STEREOTYPICAL PATTERN AND SEQUENCE OF

TOPOGRAPHICAL DISTRIBUTION AFFECTING SPECIFIC PREDILECTION SITES

252015

3

ACCUMULATION OF αααα-SYNUCLEIN IN PARKINSON rsquoS DISEASE(PD) AND PD MODELS

IS PARALLELED BY MORPHOLOGICAL EVIDENCE OF NEURONAL PATHOL OGY


RESULTS

‐‐‐‐ ‐‐‐‐H αααα ‐‐‐‐SYNUCLEIN‐‐‐‐CONTAINING AXONS OFTEN APPEARED AS

SINUOUS THREADS WITH IRREGULARLY SPACED DENSELY

LABELED VARICOSITIES

SPREADING CAUSED PATHOLOGICAL ALTERATIONS OF AXONS IN

THE PONS MIDBRAIN AND FOREFRAIN

BRAAK H ET AL (2006) NEUROSCI LETT 39667-72

AXONS APPEAR PARTICULARLY SUSCEPTIBLE TO αααα-SYNUCLEIN ACCUMULATION AND

CONSEQUENT PATHOLOGY

AXONAL PATHOLOGY HAS ALSO BEEN DESCRIBED IN THE PERIPHERALNE RVOUS

SYSTEM AT EARLY PD STAGES (IMAGE ON THE LEFT SHOWS AN AXON

IMMUNOREACTIVE FOR αααα-SYNUCLEIN IN THE GASTRIC WALL )

H αααα-SYNUCLEIN THIOFLAVIN -S MERGED

H αααα-SYNUCLEIN

‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐

STAINING WITH THIOFLAVIN ‐‐‐‐S AND CO‐‐‐‐LOCALIZATION OF

THIOFLAVIN S WITH H αααα ‐‐‐‐SYNUCLEIN IMMUNOREACTIVITY

DEMONSTRATED THAT A SMALL PERCENTAGE OF LABELED

AXONS CONTAINED Hαααα ‐‐‐‐SYNUCLEIN AMYLOID FIBRILS

bull ENHANCED PROTEIN EXPRESSION IS SUFFICIENT TO TRIGGER INTER-NEURONAL αααα-SYNUCLEIN PROPAGATION

bull OVEREXPRESSION IN THE MEDULLA OBLONGATA RESULTS IN LONG-DISTANCE CAUDO-ROSTRAL SPREADING OFαααα-SYNUCLEIN

bull αααα-SYNUCLEIN SPREADING FOLLOWS A STEREOTYPICAL PATTERN VIA ANATOMICALLY INTERCONNECTED PATHWAYS

bull αααα-SYNUCLEIN PROPAGATION AFFECTS PRIMARILY NEURONAL PROJECTIONS AND IS ACCOMPANIED BY AXONAL

PATHOLOGY

MODELING αααα-SYNUCLEIN INTER-NEURONAL PROPAGATION SUMMARY

FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

bull αααα-SYNUCLEIN ELEVATION ALSO CAUSES PROTEIN

AGGREGATION AND FORMATION OF THIOFLAVIN S-POSITIVE AMYLOID FIBERS

bull αααα-SYNUCLEIN FIBRIL FORMATION COULD

CONCEIVABLY POTENTIATE SPREADING AND NEURONAL

INJURY

MODELING αααα-SYNUCLEIN INTER-NEURONAL PROPAGATION IMPLICATIONS INCLUDE

bull THIS MODEL OF αααα-SYNUCLEIN PROPAGATION TRIGGERED

BY VIRAL VECTORS RECAPITULATES IMPORTANT

PARKINSONS DISEASE(PD) FEATURES

bull IT COULD THEREFORE BE VALUABLE FOR STUDIES ON

PATHOGENETIC PROCESSES UNDERLYING HUMAN

SYNUCLEINOPATHIES AND FOR TESTING NEW

THERAPEUTICS AGAINSTαααα-SYNUCLEIN SPREADING

FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T

bull ENHANCED PROTEIN EXPRESSION IS SUFFICIENT TO

TRIGGER αααα-SYNUCLEIN SPREADING

bull A COMMON FEATURE OF PD RISK FACTORS IS THEIR

POTENTIAL TO ENHANCE αααα-SYNUCLEIN EXPRESSION

bull PD RISK FACTORS COULD THEREFOREACT ADDITIVELY OR

SYNERGISTICALLY IN PROMOTING αααα-SYNUCLEIN

PATHOLOGY AND ITS SPREADING

Keynote Lecture Neuropathological studies- new insights

Glenda Halliday PhD Prince of Wales Medical Research Institute Randwick Australia

Neuropathology of MSA an update

Janice Holton MB ChB PhD FRCPath UCL Institute of Neurology London United Kingdom

252015

1

Neuropathology of multiple system atrophy an update

Alpha-synuclein the gateway to parkinsonismInnsbruck 11th -13th

February 2015

Professor Janice HoltonDirector of NeuropathologyQueen Square Brain BankUCL Institute of NeurologyLondon UK

Queen Square Brain Bank

Overview

bull Review the major pathological features of MSAndash Macroscopic appearancesndash Histology

bull Clinico-pathological correlations in MSAbull Discuss glial cytoplasmic inclusions in MSA

ndash Contribution to pathogenesisndash Summarise aspects of GCI formationndash Can α-synuclein mutations tell us anything about MSA


Macroscopic features

bull Atrophy and dark discolouration of the putamen

bull Cerebellar cortical atrophy

bull Atrophy and discolouration of the cerebellar hemispheric white matter with preservation of the superior cerebellar peduncle

bull Pallor of the substantia nigra

bull Atrophy of the pontine base

bull Atrophy of the inferior olivarynucleus


252015

2

Gallas silver impregnation 20-30nm tubules

Ahmed et al Neuropathol Appl Neurobiol 2011


Neuropathological subtypes of MSA

Ahmed et al 2012

1 Olivopontocerebellar atrophy (OPCA)bullAtrophy of inferior olivebullAtrophy of pontine basebullLoss of transverse fibresbullPreserved pontine tegmentum and superior cerebellar pedunclebullCerebellar cortical atrophybullDisolouration and reduced cerebellar white matter

2 Striatonigral degeneration (SND)bullAtrophy and discolouration of putamenbullPallor of substantia nigra

3 Mixed SND=OPCA


4 Minimal Change MSAbullNeuronal loss restricted to nigra locus coeruleusbullWidespread GCIs

BritishOzawa et al JNNP2010811253-1255

Ozawa et al Brain 2004 127 2657-71

Japanese

α-synuclein immunohistochemistry

Glial cytoplasmic inclusion

Glial nuclear inclusion

Neuronal cytoplasmic inclusion

Neuronal nuclear inclusion

Neuropil thread

Cellular inclusions and diagnosis of MSA

Widespread GCIs with neurodegeneration are the criteria for definite neuropathological diagnosis of MSA


252015

3

Clinicopathological correlations in MSALong duration - gt15 years (n = 4)

Long disease duration MSA vs control MSA


bull Long duration MSA may represent a more benign disease variant

Asi and Ling et al in preparation

LD-MSA vs Control MSA

GCI Caudate LD-MSAgt Control MSA

p=0002

NCI Caudate LD-MSAgt Control MSA

plt0001

Neuronal loss No significant differences

Gliosis No significant differences

Petrovic et al Mov Disord 27 1186-90 2012

bull Delayed onset of autonomic dysfunction associated with favorable clinical outcome

bull Widespread pathology in cortical limbic striatonigral and olivopontocerebellar regions

5 20Disease duration (Years)

10 15

Time to orthostatic hypotension

MSA LD

Minimal change MSA

bull Neuronal loss restricted to nigra and locus coeruleus n=6 (selected from 135 cases)

bull Controls 8 with classical disease course and progression

bull Caudate and nigra Greater NCI burden in MC-MSA

bull GCIs no difference from control MSAbull 3 with interrupted disease 3 with sudden

death (SUD)bull SUD

bull shorter disease duration 53plusmn13y vs 8plusmn33 y p=02

bull Younger onset 38plusmn40y vs 576plusmn111y p=002

bull Aggressive disease course (most milestones reached in 3 yrs)

bull Minimal change MSA may represent a more aggressive disease variant Ling and Asi et al under review



MSA cognitive impaired vs age matched MSA normal c ognition (n=9)

GC

I sco

re

GC

I sco

re

No difference inGCI burden NCI burden neuronal loss Aβ load Braak tau staging (not gtII) CAA small vessel disease

Neuropathological substrate of cognitive impairment in MSA remains to be elucidated

252015

4

r = 050 P lt 00001 r = 051 P lt 00001

Frequency of GCIs correlates with neuronal loss and disease duration implicating GCIs in neurodegeneration


Determining the mechanism of GCI formation is critical to understanding neurodegeneration in MSA

Understanding the mechanism of GCI formation


LRRK2 early in GCIs associated with p25αindicating myelin degradation

bull In MSA p25α relocates from myelin to oligodendroglial cell body

bull LRRK2 present in oligodendroglial cell body

bull Cell body becomes enlarged

bull Myelin degradation occurs

bull Subsequent deposition of fibrillar α-synuclein forming GCI

LRRK2 + P25α α-syn

Wenning et al Ann Neurol 2008 64 239-246

Huang et al Acta Neuropathologica 2008



Initiating factorsNeuroinflammation

SourceSynthesised by oligosFrom neurons

Genetic influences in MSA

Altered structureInfluence on fibril formationInfluence on protein degradationInfluence on cell-cell transmission



252015

5

Microglia in MSA

MSA Control


Microglial burden is increased in the white matter in MSA in mildly and

severely affected regions

First demonstration of α-synuclein mRNA by qPCR in oligos in postmortem tis sue


Source of α-synuclein ndash oligodendrocytes

Cellular expression (LCM) qPCR

Neurons Oligodendrocytes

α-Synuclein mRNA is expressed in neurons and oligos

Hansen and Li Trends in Molecular Medicine 2012 18

PD Braak hypothesis cell-to-cell transfer of α-syn

Alpha synuclein

Angot et al Lancet Neurology 2010 9 1128

MSA

Neuron

Oligodendrocyte


252015

6

Recombinant monomeric and oligomeric α-synuclein taken up by oligodendrocytes

In Vitro

Mon

omer

Olig

omer

In Vivo

In rat model expressing human α-synuclein in nigral neurons and axons demonstrated transfer of α-synuclein from neurons into grafted OPC and mature oligodendrocytes


Insular cortex

Caudate Substantia nigra

Case Case one Case two Case three Age of onset (years)

19 69 46

Disease duration (years)

29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor

Final clinical diagnosis

Familial pallidopyramidal

syndrome

Parkinsonrsquos disease with

dementia


dementiaLevodopa responsive

Good and sustained

Transient Transient

Cognitive impairment

Yes Yes frontal predominant

Yes frontal predominant

Visual hallucinations

Yes Yes Yes

Autonomic dysfunction

Yes Yes Yes

Pyramidal signs Yes Yes Yes

Family history of parkinsonism

Father sister Mother aunt brother son

Mother uncle grandmother

α -synuclein α -synuclein

OligoNeuronal


SNCA

Mutation

α-synuclein pathology

Pattern Glial

Golbe 1990 Duda

2002A53T PD-like

Spira 2001 A53T PD-like None

Markopoulou 2008 A53T PD-like Few GCI

Seidel 2010 A30P PD-like PD type

Zarranz 2004 E46K PD-like

Obi 2008 Duplication PD +MSA Few GCI amp CB

Kara 2014 Duplication PD-like PD type

Ikeuchi 2008 Duplication PD-likeFew type

not specified

Gwinn-Hardy 2000 Triplication PD +MSA Few GCI amp CB

Farrer 2004 Triplication PD-like

Proukakis 2013 H50Q PD None

Kiely 2013 G51D PD +MSA GCI and CB

Pasanen 2014 A53E PD +MSA GCI


Kiely et al in preparation

Over production or mutation of α-syn may predispose to GCI formationMutation structural alteration altered fibril formation kinetics or impaired degradation

Kara et al JAMA Neurol 2014

cortex

SN cortex

252015

7

Conclusions

bull Clinicopathological studies ndash Long disease duration and late autonomic failure - benign disease sub-typendash Minimal change MSA - more aggressive phenotype younger onsetndash Cognitive impairment in MSA is not related to α-synuclein load and distribution or to

other concomitant pathologies

bull α-synuclein containing GCIs are important in MSA pathogenesis

bull There are changes in oligodendrocytes before the aggregation of α-synuclein ndash understanding triggers could be important

bull Oligodendrocytes mayndash Synthesise α-synucleinndash Take up α-synuclein from neurons

bull Altered biochemical properties of α-synuclein may be important in GCI formation ndash SNCA mutations may be informative


α -synuclein

Oligo

Queen Square Brain BankJanice HoltonTamas ReveszAndrew LeesTom WarnerZeshan AhmedYasmine AsiAoife KielyHelen LingIgor PetrovicTammaryn LashleyCatherine StrandRobert CourtneyAbi LiChristina MurrayKaren ShawLinda ParsonsSusan StonehamIliyana Komsiyska

UCL Institute of Neurology

Clare FowlerNiall Quinn

Dept of Molecular NeuroscienceHenry HouldenAnna SailerJohn HardyLucia SchotlaenderNick Wood

Brain donors their families and carers

National Hospital for Neurology and Neurosurgery

Niigata UniversityTetsutaro Ozawa

FundingThe Multiple System Atrophy TrustThe Reta Lila Weston InstituteKuwait Ministry of Higher Education

Sheffield UniversityJulie SimpsonPaul HeathStephen Wharton


New insights into pathomechanisms

Nadia Stefanova MD PhD Medical University of Innsbruck Innsbruck Austria

252015

1

MSA New insights into

pathomechanisms

Nadia Stefanova MD PhD



Alpha-synuclein The Gateway to Parkinsonism

Multiple System Atrophy (MSA)

The American Journal of Pathology (2007) 171(4)

Brain (2008) 131

J Neuropathol Exp Neurol (2004) 63(1)Neurology (2003) 61(5)

Lancet Neurol (2004) 3(2)

Glial cytoplasmic inclusions (GCIs) Early TPPPp25 translocation

Neuroinflammation

PD

MSA

Genetic predisposition

Oligodendroglial α-synuclein in MSA

bull Intracellular source in

oligodendrocytes

ndash pros

ndash cons

bull Extarcellular source

α-synuclein transmission to

oligodendrocytes experimental

evidence

Asi et al 2014

Solano et al 2000 Miller et al 2004

Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2


-intracellular overexpression-MBP-aSYN transgenic mice

bullAstrogliosis

bullDemyelination

bullAxonal alterations in neocortex

basal gangliabrain stem

cerebellum

bulldarr TH fibers in striatum

bullNo TH loss in SNc

bullMotor disability

bullPreserved autonomic phenotype

PLP-aSYN transgenic mice

bullMicrogliosis

bullAxonal alterations


bull TH loss in SNc

bullNeuronal loss in Namb LDT PPT

bull Neuronal loss in Onuflsquos nucleus


bullCardiovascular dysfunction

bullBladder dysfunction

bullPreserved olfaction

αSYN

phospho-αSYN

Shults 2005 Ubhi 2008 2009 2010 Tank 2014 Kahle 2002 Stefanova 2005 2007ab 2008 2009 2011ab 2013 Stemberger

2010 Kuzdas 2013 Boudes 2013 Krismer 2013 Kuzdas-Wood 2014

bullGCIs bullGCIs


-intracellular overexpression-

Reduced trophic support

bull darrGDNF

Epigenetic dysregulation

bull darrH3 acetylation

Ubhi 2010

bull H3 acetylation is reduced in MSA mouse brains

bull NaPB protects nigral dopaminergic neurons in MSA mice

Sturm in preparation



Neuroinflammation

bull Role of TLR4 on microglia in the clearance of α-synuclein

bull Detrimental role of uarriNOS and uarrMPO in microglia

Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3

Increased susceptibility to environmental stress

in oligodendroglial α-synucleinoapthy

Proteasome inhibition Oxidative stress

Striatonigral degeneration Olivopontocerebellar atrophy

Astrogliosis Microgliosis

Motor disability

Stefanova 2005

Striatonigral degeneration

Olivopontocerebellar atrophy

Motor disability

Stefanova 2011

Summary oligodendroglial α-synuclein overexpression may

lead to neurodegeneration

Mechanisms

bull Reduced trophic support

bull Epigenetic dysregulation

bull Disbalance of neuroinflammatory responses

bull Lower threshold of suscepibility to oxidative and proteolytic stress


-uptake of extracellular protein-

Is prion-like propagation relevant to MSA pathogenesis

Watts 2014

Reyes 2014

252015

4

Oligodendroglial α-synucleinopathy and

mechanisms of neurodegeneration in a

glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines

α-synuclein clearance

Preclinical target discovery

Poul Henning Jensen MD University of Aarhus Aarhus Denmark

252015

1

Poul Henning JensenAlpha-Synuclein The Gateway to Parkinsonism 2014

Multiple system atrophy


An enigmatic disease


2

Oligodendroglial cytopathology before and after AS enters

the stage in MSA

Pre Post-α-synuclein GCI phenotype


Novel in vivo observationsLipid metabolism in MSA

bull ABCA8 is a sphingolipid transporter

expressed in white matter

bull Expression increased in MSA

bull Transgenic ABCA8 expression in

oligodendroglial cell line increases

p25a expression (Kim 2013

Biochem J Bleasel 2013 J Park

Dis)

Nuclear p25a

bull A novel characteristic for stratifying

oligo-pathology which is inversely

correlated to oligodendroglial MSA

cytopathology (Ota 2014 Acta

Neuropath Comm)

252015

2


Preclinical target discoveryInvestigating the post-α-synuclein GCI phenotype

bull Inhibitor of SIRT2 acetyl transferase protect cells and increase AS inclusionsaggregates in a-synp25a tg cell model (Hasegawa 2010 Neurochem Int)

bull FAS blockade protects in a-synp25a tg cell model and Plp-a-syn expressing primaryoligodendrocytes (Kragh 2013 PloS One)

bull TNF blockade protects in a-synp25a tg cell model (Kragh amp Jensen unpublished)

bull IkBia expression is increased in a-synp25a tg cell model and MBP-a-syn tg micelines and IkBia silencing protects cell model (Kragh 2014 Neurobiol Dis)

Oligo-neuron interplay in mixed primary cultures of CNP-asyn tg mice

bull Cystatin C is released from a-syn expressing oligos and mediate neuronalaccumulation of endogenous AS and apoptosis mediate toxicity (Suzuki 2014 Am J Pathol)

bull Oligo a-syn induce neuronal a-syntubulin dependent neurodegeneration that canbe rescued by small tubulin peptide (Suzuki 2014 JBC)

MSA brain extract triggers prion-like degeneration in M83 a-syn tg mice (Watts 2013 PNAS)


Facts and loose ends

bull Neurons die ndash but before or after oligodendrocyte engagement

bull Does oligodendrocytes die to what extend (flux) and are they

replenished

bull Is myelin relevant given non-myelinating cortical oligodendrocytes

also develop GCI

bull Can we divide the disease in a yet unknown triggering phase and a

neurotoxic executioner phase driven by AS

bull Which part of the strong tissue reactivity is driving and being

driven by the etiopathogenic process Role of micro- and astroglial

activation involvement of peripheral factors due to breakdown of

blood brain barrier


Find the known unknown and maybe

the unknown unknown

Need for close collaboration

between experimentalist and

pathologist

Table of Contents

Course Description and DisclaimerPage 3 Venue and Accreditation InformationhelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellipPage 4 Faculty Listing and Industry Disclosureshelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellip Page 5 Agenda February 11 2015 helliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellipPage 9 Agenda February 12 2015 helliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellipPage 9 Agenda February 13 2015 helliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellipPage 10 Session 5 Abstract Listinghelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellip Page 11

Faculty PresentationshelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellipPage 16

2






Learning Objectives




Evaluations


3



Course Venue




4

Course Directors



Course Faculty


5


6



7


8

Scientific Program





9






10


1



2



3



4



5



6



7



8



9




11


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist






Learning Objectives




Evaluations


3



Course Venue




4

Course Directors



Course Faculty


5


6



7


8

Scientific Program





9






10


1



2



3



4



5



6



7



8



9




11


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

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ity(d

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m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

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liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

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liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

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liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist



Course Venue




4

Course Directors



Course Faculty


5


6



7


8

Scientific Program





9






10


1



2



3



4



5



6



7



8



9




11


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

Course Directors



Course Faculty


5


6



7


8

Scientific Program





9






10


1



2



3



4



5



6



7



8



9




11


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


6



7


8

Scientific Program





9






10


1



2



3



4



5



6



7



8



9




11


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist



7


8

Scientific Program





9






10


1



2



3



4



5



6



7



8



9




11


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


8

Scientific Program





9






10


1



2



3



4



5



6



7



8



9




11


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

Scientific Program





9






10


1



2



3



4



5



6



7



8



9




11


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist






10


1



2



3



4



5



6



7



8



9




11


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


1



2



3



4



5



6



7



8



9




11


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


11



12



13



14



15



16



17



18



19



20



12



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist



22



23





25



26



27



28







31




13


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


33



34



35



36



37



38



39



40



41



42




14


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


45



44



45



46



47



48



49



15


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


16



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist



252015

1



Sensitive


Inexpensive






3

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

2










neurological signs

medulla oblongata

pontine tegmentum





SN IV


SN IV

Aurbach V


252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

3









252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

4



Synuclein Ubiquitin

Synuclein Ubiquitin

Host

nig

raG

raft

ed n

euro

ns



8006 Japanese



252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

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200 Wavelength (nm)

220 240

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Mea

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liptic

ity(m

deg

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mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

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inte

nsity

x 1

0-3

0

2

4

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[α-syn][DMPS]

15

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40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

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inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

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ate



a

[DMPS][α-syn] (MM)

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10-3

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-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

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inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

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inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

5


Bowel

movements


cases

Unadjusted Age-

adjusted

lt 1 289 10 796 189

1 4371 66 80 79

2 1704 17 52 54

gt2 426 3 38 39


Overall 6790 96 75






local








252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

6

Agesex Disease

Duration

Onset dx (y)

tUPDRS HY BMd

55M 425 28 2 1

66M 15 27 2 1

56M 175 15 15 lt1

78M 15 20 2 1

75M 41 28 2 lt1

68F 0533 24 2 2

46F 21 16 1 lt1

49M 812 18 2 1

61M 15 18 2 1

51M 215 28 2 1-2

585 1575 22 2Median


Age median(range) 54 (36-71) 40 (24-72) 43 (21-62)

Gender male 12 (46) 6 (55) 4 (36)

Parkinson disease

Crohnrsquos disease

Control

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

7


A

C

E

D

F

B

Parkinsonrsquos

Ulcerative

Colitis


A

C

B

D



MCI

Rest tremor 22010


MPTP MPTP

Aged Aged

Young

control

Young

control

low

low

low

high

high

high

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

8

Alpha syn TH Merged

Sugar Site

Sucrose Gastric


Sucralose colonic


0

05

1

15

2

25

control PD

p=013

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

9

LBP (ngml)

0

50000

100000

150000

200000

250000

300000

control PD

p=0016




Gram (ndash) rods

bacteroides

desulfovibrios

escherichia

fusobacteria

Gram (+) c0cci

ruminococci

peptostreptococci

peptococci

streptococci

Gram (+) rods

eubacteria

bifidobacteria

clostridia

lactobacilli

propioibacteria

actinomyces


methanobrevibacter


252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

10





ribotyping (RNA

polymorphisms)

reliable high


culturable only



culturable

costly


Parkinson disease

(N=10)

Crohnrsquos disease

(N=11)

Ulcerative colitis

(N=13)

Controls

(N=26)

Age median

(range)

57 (46-79) 40 (24-72) 42 (21-62) 55 (36-71)


Duration PD (y)

Median (range)

15 (05-8)


HY Stage

III

28

Healthy Lumen

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 50

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8

Parkinson lumen

Healthy sigmoid

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 5 6 7 8 9

Parkinsons sigmoid

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

11


Control lumen

Control mucosa

Parkinson mucosa

Parkinson lumen

Axi

s 2

Axis 1

-01

-02

-03

-04

-05

01

02

03

04

-01-02-03-04-05 01 02 03 04

All Parkinson

Agesex Disease

Duration

(y)


lumen

55M 4 28 2 1 ++ ++ radic radic

57M 1 27 2 1 ++ ++

48M 1 15 15 lt1 ++ ++ radic radic

77M 1 20 2 1 ++++ ++++



60M 1 18 2 1 ++++ ++++ radic

57M 2 28 2 1-2 ++ ++++ radic

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

12

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

13





Christopher Forsyth




252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist



252015

1










bull Hyposmia

bull Sleep



bull Otherbull pain


Lim amp Lang 2009


252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

2




2004

ThSG

ThSG

FrontCtx

SNpc

CingulCtx

SNpc









252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

3








bull Dizziness






bull Gluteal ache

bull Calf ache

Renal hypoperfusion

bull Oliguria

Non-specific

bull Fatigue



2003

2006

14 (7 51)

Early 60 (21 35) Premotor 11 (4 35)


252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

4




dysfunction and

constipation could




from 50 to 90






MDJ 2012

Premotor OH






252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

5




2013)


2010 Jain 2012 Palma 2013)




MDJ 2012PDHCMSA



MDJ 2013

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

6








Urinary biomarkers

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

7


Feature MSA PAF PD











supersensitivity


Garland et al 2013



Conclusions








252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

8


Acknowledgements





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist





Introduction




Review of Evidence

Postuma 2012

DLB versus PDD






Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist





Redefining PD


0

5

10

15

20

25



Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


Maetzler 2009


Braak 2003


another


Ravina 2012




Duncan under review


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


Hall 2015


Stewart 2014


Zokaei 2014


Zokaei 2014



Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


Gagnon 2009


PD-RBD

Patients

PD-NRBDPatients

ControlSubjects

N 32 22 18 40

MCI n () 16 (50) 16 (73) 2 (11) 3 (8)


2 (125) 0 0 0







1 4 0 0


0 0 0 0




2 (125) 3 (19) 0 0



Berg 2011

Conclusion
























patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist





















patrikbrundinvaiorg







12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist





12022015











Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist









Our approach







-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist



-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b





-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist



-30 -25 -20 -15 -10 -5 0 5

Mea

n re

sidu

e el

liptic

ity(d

egc

m2 d

mol

-1) x

10-3

200 Wavelength (nm)

220 240

a

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

-30

-20

-10

0

-5

-15

-25

0 1 2 3[DMPS] (mM)

b222 nm a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10N

orm

alis

ed m

axim

um r

ate



a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


a

[DMPS][α-syn] (MM)

MR

E x

10-3

(deg

cm

2 dm

ol-1

)

-25

-20

-15

-10

-5

0

0 10 20 30 40 50 60


b

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

0

2

4

6810

[α-syn][DMPS]

15

c d

40

30

20

[α-syn][DMPS]

Time (h)0 20 40 60

0

1

2

3

Fluo

resc

ence

inte

nsity

x 1

0-3

15

0 10 20 30 40 50 600

02

04

06

08

10

Nor

mal

ised

max

imum

rat

e



Fluo

resc

ence

inte

nsity

(au

)

0

200

400

600

0 50 100Time (h)





b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist


b

Mea

n re

sidu

e el

liptic

ity(m

deg

cm2 d

mol

-1) x

10-3

Wavelength (nm)

-30190 200 210 220 230 240 250

-20

-10010

20

3040

5060 0

-5-10-15-20-250 1 2 3 4 5

MR

E x

10-3

Time (h)

a

Time (h)

Fluo

resc

ence

inte

nsity

x 1

0-3

00

4

2

6

8

10

12

14

5 10 15 20


842








0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist





0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d




0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist



0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d

0

1

2

3

4

5

6

7

60 100 140 180020406080

[α-s

yn] fib

rils (μ

M)

[α-synfree] (μM)

Fluo

resc

ence

inte

nsity

x 1

0-3


[α-synfree](μM)2001751251008060

Time (h)

0

1

2

3

4

5

0 20 40 60 80 100 120

a

Fluo

resc

ence

inte

nsity

x 1

0-3

6

Time (h)0 20 40 60 80 100 120

[α-synbound](μM)


2468

1015

20

40b

80 120 160 200

100120140160180200

0

20

40

60

80

[α-s

yn] fib

rils (μ

M) 100

120

140c

0 10 20 30 40[α-synbound] (μM)

d





















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist




















Methods 2011)






Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist




Conclusions






Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

Acknowledgements

Thomas CT Michaels


Dobson









Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist




Seung-Jae Lee







252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist





252015

1



AYSEULUSOY

MICHAEL HELWIG

RAFFAELLA RUSCONI

MICHAEL KLINKENBERG

RUTH MUSGROVE


FRANZISKA HESSE

ACKNOWLEDGEMENTS






RN

A( αα αα

-SY

NU

CLE

INS

YN

AP

TOP

HY

SIN

)

1

0

2

CONTROLS PATIENTS


179










EXPERIMENTAL MODELS


TH





ANIMALS (EG RATS)



252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

2




THE RAT NECK

1





3

3

LOWER BRAIN

4

1

VAGUS NERVE

2





4








IMM

UN

OR

EA

CT

IVIT

YFO

RH

UM

AN

αα αα-S

YN

UC

LEIN


DMV NA




RESULTS

18 W

EE

KS

RMB

FB


8 W

EE

KS

PONS

CMB


4 W

EE

KS

MO


PREDILECTION

SITES



DORSAL RAPHAE

PONS

MIDBRAIN

FOREBRAIN





252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

3




RESULTS














‐‐‐‐ ‐‐‐‐

‐‐‐‐

‐‐‐‐









PATHOLOGY


FIBRILLATION

PROTEIN LOAD

INJURY SPREADING





INJURY









FIBRILLATION

PROTEIN LOAD

INJURY SPREADING

GENES

AGING ENVIRONMEN

T












252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist





252015

1



February 2015



Overview













252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

2





Ahmed et al 2012



3 Mixed SND=OPCA





Japanese






Neuropil thread




252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

3








p=0002


plt0001







10 15


MSA LD

Minimal change MSA





death (SUD)bull SUD








GC

I sco

re

GC

I sco

re



252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

4

r = 050 P lt 00001 r = 051 P lt 00001























252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

5

Microglia in MSA

MSA Control












Alpha synuclein


MSA

Neuron

Oligodendrocyte


252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

6


In Vitro

Mon

omer

Olig

omer

In Vivo



Insular cortex



19 69 46


29 6 6

Presenting symptoms

Resting hand tremor

Resting hand tremor

Resting hand tremor



syndrome


dementia



Good and sustained

Transient Transient





Yes Yes Yes


Yes Yes Yes






OligoNeuronal


SNCA

Mutation


Pattern Glial

Golbe 1990 Duda

2002A53T PD-like








not specified










cortex

SN cortex

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

7

Conclusions








α -synuclein

Oligo













252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist



252015

1


pathomechanisms







Brain (2008) 131




Neuroinflammation

PD

MSA




oligodendrocytes

ndash pros

ndash cons




evidence

Asi et al 2014


Reyes et al 2013

CNP + αSyn

Fellner unpublished

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

2



bullAstrogliosis

bullDemyelination



cerebellum






bullMicrogliosis



bull TH loss in SNc







αSYN

phospho-αSYN



bullGCIs bullGCIs




bull darrGDNF



Ubhi 2010






Neuroinflammation



Stefanova 2007

Stefanova 2011

Stefanova 2011

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

3






Motor disability

Stefanova 2005



Motor disability

Stefanova 2011



Mechanisms








Watts 2014

Reyes 2014

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

4



glance

Proteolytic

stress

Oxidative stress

GCI

Pro-inflammatory

cytokines




252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist



252015

1






2


the stage in MSA











Dis)

Nuclear p25a





Neuropath Comm)

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

252015

2















replenished


also develop GCI






blood brain barrier



the unknown unknown



pathologist

alpha-synuclein: the gateway to parkinsonism- innsbruck

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