forschungstag der universitäts-kinderkliniken bern · resting state functional connectivity and...
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Forschungstag der Universitäts-Kinderkliniken Bern
16. Oktober 2019
Kuppelraum
Hauptgebäude
Universität Bern
Forschungstag der Universitäts-Kinderkliniken Bern
Datum
Mittwoch, 16. Oktober 2019
Ort
Kuppelraum, Hauptgebäude, Universität Bern
Organisation
Birgit Seyfried
Forschungssekretariat
Universitätsspital für Kinderheilkunde
Inselspital, Universitätsspital Bern
Tel: +41 31 632 94 93
E-Mail: [email protected]
9:00Begrüssung
Christa Flück IT-Support, Florian Singer
Orale Präsentationen 1, Neurologie
Chairs: Karen Lidzba, Rahel Kasteler
9:15
Janine Spitzhüttl
Impact of Non-CNS Childhood Cancer on Resting-State Connectivity
and its Association with Cognition
9:27
Leonie Steiner
Resting state functional connectivity and motor functions in patients
with and without hemiparesis after pediatric arterial ischemic stroke
9:39
Celine Hochstrasser (geb. Webland)
A new test for assessing processing speed in children: Swiss validation of
the c-SDMT
9:47Quendresa Thaqi
The impact of age at arterial ischemic stroke on executive functions
9:55
Robin Münger
Focal cerebral arteriopathy in childhood stroke of the posterior circulati-
on – analysis of a population based registry
10:02
Nicola Biesold
Hypoxic-ischemic encephalopathy in adolescents; neuropsychological
impairments and rehabilation outcome - a case report.
10:09
Daniel Brechbühl
Isolated pediatric Neurosarcoidosis presenting as bilateral subacute
complete sensorineural hearing loss
10:16
Cornelia Enzmann
A cross-sectional and longitudinal natural history study of the Swiss
cohort of LAMA2-related congenital muscular dystrophy
10:23Valerie Siegwart
Cognition, behavior, and quality of life in pediatric cancer survivors
10:45 Kaffeepause
Programm Forschungstag der Universitäts-Kinderkliniken Bern
Orale Präsentationen 2, Onkologie
Chairs: Eva Brack, Michele Bernasconi
11:00
Luana Lavieri
Risk prediction of fever in neutropenia in children and adolescents under
chemotherapy for cancer in a prospective, multicenter trial
11:12
Mutlu Kartal-Kaess
High throughput sequencing of CD34+ selected peripheral blood stem
cells in children with various cancer types undergoing autologous stem
cell transplantation
11:24Lukas Meyer-Landolt
Paraneoplastic pruritus in patients with Hodgkin Lymphoma Case report
11:31
Christa König
Continuous monitoring of health data with a wearable device in
pediatric patients undergoing chemotherapy for cancer – a feasibility
pilot study
11:38
Dzhangar Dzhumashev
Screening of peptide ligands for targeted drug delivery to rhabdomyos-
arcoma cells
11:45Andrea Timpanaro
Surfaceome profiling of rhabdomyosarcoma for CAR T cell targets
11:52
Bettina Blank
Challenges of treating osteosarcoma in a 12-years old patient with Sickle
Cell Disease
12:00
Elizaveta Fasler-Kan
Gastrin releasing peptide receptor is overexpressed in Wilms tumour
patients and cell lines
12:15 Lunch
13.30 Gastreferat: Prof. Claudia Kuehni
“Use of hospital routine data for research? Limitations and opportunities”
Short biosketch
Prof. Claudia Kuehni: Medical studies in Bern. Training in paediatrics (FMP Paediatrics) with
subsequent specialisation in paediatric pulmonology in St.Gallen, Delémont and Bern). MSc
in Epidemiology in London (LSHTM, 1996-7), research fellow in Leicester, UK (1997-99).
Current position as research group leader and associate Professor in Paediatric Epidemiolo-
gy at the Institute of Social and Preventive Medicine at the University of Bern.
Claudia Kuehni is an expert in the epidemiology of childhood diseases, especially respirato-
ry diseases, childhood cancer, and rare diseases. Her research focuses on the causes,
treatments and long-term course of common and rare diseases in childhood. She leads
SwissPedRegistry (the registry hub of SwissPednet), and national or multicentre cohort
studies and registries (Leicester Respiratory cohort studies, Swiss Childhood Cancer
Registry, Swiss Rare Disease Registry, Swiss Paediatric Airway Cohort (SPAC), Swiss Primary
Ciliary Dyskinesia Registry, iPCD cohort and many more), and is responsible for the
surveillance of the neonatal screening for Cystic Fibrosis.
Claudia is a member of the research council of the Swiss National Science Foundation
(Division 3), Paediatric section editor for the European Respiratory Journal, and member of
many other boards relevant for research in Child Health.
Orale Präsentationen 3, Pneumologie und Physiotherapie
Chairs: Christiane Sokollik, Florian Singer
14:30
Johanna Kurz
Association of lung clearance index and survival in patients with cystic
fibrosis
14:42
Ruth Stauffer-Lacorcia
Improving opportunities for physical activity and participation in
inpatient settings for adolescent cancer patients
14:54
Bettina Frauchiger
Lung clearance index: a promising biomarker to track lung disease
progression during childhood in patients with cystic fibrosis
15:06
Marc-Alexander Oesterreich
Systematic differences in analysis software versions hamper infant lung
function testing
Orale Präsentationen 4, Gastrologie & Endokrinologie/ Metabolik
Chairs: Christa König, Claudia Böttcher, Matthias Gautschi
16:00
Mariia Borsuk
Lack of aquaporin 9 expression in iPSC-derived hepatocytes impairs urea
secretion
16:12
Shaheena Parween
Effect of metformin on ACTH receptor activation and downstream
signaling
16:24
Mara Grassi
Expanding the phenotype of the p.R63W mutation of HNF4A – a case
report
16:31Emanuele Pignatti
New Mechanistic Insights into Androgen Production
16:38
Tanja Zingg
Level of glycemic control in pediatric patients with type 1 diabetes in
Bern: a cross-sectional study
16:45
Luca Cecchini
Trajectory of microbiota maturation in healthy Bern infants - a network
approach
16:52
Efstathios Katharopoulos
Human-induced steroidogenic cells from urine consist a new cell-based
model to study adrenal disorders in a personalised manner
17:00 Schlusswort und Preisverleihung, anschliessend Apéro
15:13
Andras Soti
Join Effects of prenatal smoke exposure and polymorphism in the
MBL2-gene on cord blood levels of mannos-binding lectin
15:30 Kaffeepause
Resting state functional connectivity and motor functions in patients with and
without hemiparesis after pediatric arterial ischemic stroke
Leonie Steiner1*, Stephanie Winkelbeiner2,3*, Andrea Federspiel2, Salome Kornfeld1,4,
Regula Everts1, Sandeep Kamal1, Juan Delgado Rodriguez1, Nedelina Slavova5, Roland
Wiest5, Maja Steinlin1, Sebastian Grunt1
* The authors contributed equally to this work
1 Division of Neuropaediatrics, Development and Rehabilitation, Children’s University
Hospital, Inselspital, Bern, Switzerland.Division of Systems Neuroscience
2 Translational Research Center, University Hospital of Psychiatry and Psychotherapy,
Bern, Switzerland
3 Center for Psychiatric Neuroscience, The Feinstein Institute for Medical Research,
Manhasset, NY, USA
4 Center for Cognition, Learning and Memory, University of Bern, Bern, Switzerland
5 Institute of Diagnostic and Interventional Neuroradiology, University Hospital, Inselspi-
tal, University of Bern, Bern, Switzerland
Introduction: The objective of this cross-sectional observational study was to investigate
the relationship between brain functional connectivity and severity of sensorimotor
impairments in the upper limb in the acute phase post stroke.
Patients and Methods: Patients with arterial ischemic stroke (diagnosed <16 years, >2 years
after diagnosis) and age-matched healthy controls were recruited from the Swiss Neurope-
diatric Stroke Registry. Eight patients with hemiparesis, ten patients without hemiparesis
and thirteen healthy controls underwent and a clinical sensorimotor assessment and
resting-state functional magnetic resonance imaging (rs-fMRI). Functional connectivity of
the motor network -including the primary motor cortex (M1), dorsal premotor cortex
(PMC), supplementary motor area (SMA), prefrontal cortex (PFC) and the superior parietal
lobus (SPL) - was assessed.
Results: The three groups displayed different functional connectivity indices in terms of
interhemispheric and intrahemispheric connectivity (p = 0.001). Moreover, significant
associations were found between sensorimotor outcome and interhemispheric (r = -0.63)
and intrahemispheric (r = -0.51) network indices.
Conclusion: We found different network patterns in patients with hemiparesis, patients
without hemiparesis and healthy controls in the motor network. These findings can help to
better understand the mechanisms of motor recovery. They are essential for a better
understanding of how targeted therapies after childhood stroke may affect network
changes.
A new test for assessing processing speed in children: Swiss validation of the
c-SDMT
Céline Hochstrasser, Ursina Jufer, Li Mei Cao, Marie-Noëlle Klein, Sarah Rieder, Michelle
Steiner, Sandra Bigi, Karen Lidzba
Neuropediatrics
Background: Processing speed (PS) is a marker for cognitive function in patients with
neurological conditions. Associated with neural maturation, it increases during develop-
ment. Traditionally, PS is measured by paper-pencil tasks requiring adequate fine motor
skills, which are often impaired in patients with neurological conditions. Thus, a non-motor
alternative is desirable. The Candian computerized Symbol-Digit Modalities Test (c-SDMT)
requires the patient to verbally associate numbers with symbols. The examiner measures
speed via key-press. It has been standardized in a large Canadian paediatric sample and
validated in patients with paediatric MS.
Method: 86 neurologically healthy participants (8 - 16 years; 45 male; 48 hospitalized for
non-neurological reasons) were examined from 06/18 – 11/18, 48 of these returned for a
retest two weeks later. All participants performed the written and computerized SDMT and
a nonverbal intelligence screening (TONI-4). Statistical analyses included a comparison of
the Swiss data to the Canadian norms, a multiple regression on the effects of age and
hospitalization on the performance on the (c-)SDMT, ANCOVAs for the effects of age, sex
and IQ, as well as measures of test-retest reliability.
Results: 1. The Swiss data is comparable to the Canadian norms. 2. As expected, age is the
best predictor for performance on (c-)SDMT. 3. Hospitalization influences the performance
on the SDMT but not on the c-SDMT. 4. Sex and IQ do not have an effect. 5. The (c-)SDMT
have an excellent test-retest reliability, however, practice effects are high and have to be
taken into account.
Conclusions: The c-SDMT is a valid and reliable measure of processing speed in children
and adolescents. The Canadian norms can safely be used to evaluate the performance of
Swiss children. The c-SDMT is more feasible for the use in hospitalized populations than the
SDMT. In future studies, we will validate the c-SDMT in selected neurological populations.
The impact of age at arterial ischemic stroke on executive functions
Thaqi, Q.1,2,3, Steinlin, M.1, Roebers, C.2,3, & Everts, R.1,2
1 Division of Neuropediatrics, Development and Rehabilitation, Children’s University
Hospital, Inselspital, University of Bern, 3010 Bern, Switzerland;
2 Swiss Graduate School for Cognition, Learning, and Memory (GSCLM), University of
Bern, 3012 Bern, Switzerland;
3 University of Bern, Department of Psychology, 3012 Bern, Switzerland
The development of executive functions (EF) is closely related to the maturation of the
prefrontal cortex, which is particularly vulnerable to brain injury due to its prolonged
development. EF, such as the ability to inhibit prepotent responses, flexibly shift between
mindsets, and store and manipulate information in short-term memory, have rarely been
investigated following a pediatric arterial ischemic stroke (AIS).
The current study investigated in a cross-sectional study design how the age at AIS
influences the development of EF, namely inhibition, shifting, working memory, and
processing speed after AIS (age at assessment: M = 15.4, range = 8.3 – 22.7; neonatal
stroke < 28 d, early childhood stroke < 7 yr, late childhood stroke > 7 yr).
There was no significant difference between age at stroke groups in fluid intelligence.
Overall, mean values of EF in all groups of patients after AIS were within the normal range.
Patients after neonatal stroke showed significant better shifting and working memory
performance than patients after late childhood stroke. Exploratively, mean values for EF and
processing speed were slightly better in patients after neonatal stroke than in patients after
early and late childhood stroke, except for errors in inhibition and errors in shifting.
The findings in the present study propose that age at AIS has an impact on cognitive
functions. More specific, our results show that a neonatal stroke has a benign impact on
cognitive functions which are not yet initiated at the time of brain injury (i.e. on EF). Thus,
our data point towards better outcome, when AIS occurs before the initiation and establish-
ment of EF development. Shifting and working memory are developing during early and
late childhood. AIS during this critical period of EF development seems to have a more
detrimental impact on outcome.
Focal Cerebral Arteriopathy of Children in Posterior Circulation – Analysis of a
population based registry
Robin Münger, Nedelina Slavova, Iciar Sanchez-Albisua, Maja Steinlin
Department of Pediatric Neurology, University Children’s Hospital, University of Bern,
Switzerland (R.M., I.S., M.S.,) and Department of Neuroradiology, Bern University
Hospital, University of Bern, Switzerland (N.S.).
Background and Purpose: Focal cerebral arteriopathy (FCA) is an important risk factor for
arterial ischemic stroke (AIS) in childhood, known especially in the anterior circulation. AIS
due to FCA in the posterior circulation is less well known and scarcely described. This
population based cohort study aimed to define the incidence of AIS due to FCA-i (infectious
triggered) in the posterior circulation and to compare them to anterior circulation cases.
Methods: We reviewed all cases of childhood AIS (age 1 month to 16 years) in the Swiss
Neuropediatric Stroke Registry from January 2000 until December 2018, with abnormal
magnetic resonance angiography, recent acute Varicella infection, or the presumption of a
vasculitis to identify FCA-i cases. Initial clinical presentation provoking factors, MR imaging
findings and outcome by Pediatric Stroke Outcome Measure (PSOM) were compared
between anterior and posterior cases.
Results: We identified 67 cases of FCA in the SNPSR, with eighteen posterior cases (27%),
resulting in an incidence of 0.08/100 000 (95% CI 0.039 to 0.112) for posterior and
0.28/100 000 (95% CI 0.248-0.317) for all strokes due to FCA-i.
Median age at presentation was significantly different with 7.9 years in the posterior group
and 5.4 years in anterior group. Further significant differences were found in the association
with recent varicella infections (16.7% vs. 49%, p 0.024), clinical manifestation as facial
palsy (22.2% vs. 73.5%, p 0.0002), headache (72.2% vs. 22.5%, p 0.0003), ataxia (61.1%
vs. 14.3%, p 0.0003) and nausea and vomiting (61.1% vs. 20.4%, p 0.003).
Conclusion: Children presenting with posterior stroke due to FCA-i are older, present with
more subtle symptoms but do not differ in outcome. One fourth of FCA-i cases affected the
posterior circulation, this points to the importance to include posterior FCA-i in differential
diagnosis and in future research.
Hypoxic-ischemic encephalopathy in adolescents; neuropsychological impair-
ments and rehabilitation outcome - a case report.
Nicola Biesold (M. Sc. Psychology), Co-Author: Prof. Dr. K. Lidzba
Neuropediatrics, Development and Rehabilitation
This case study investigates the relationship between brain scans and neuropsychological
impairments of patients with hypoxic-ischemic encephalopathy (HIE) and the importance
of multidisciplinary therapies.
HIE, also known as hypoxic brain injury is caused by deprivation of oxygen to the brain.
Thorax traumas with hypoxic encephalopathy in children are very rare, normally caused by
accidents. Yet, professional treatment in the first minutes is crucial for survival and
outcome. Otherwise the grey matter, the most vulnerable structure, can be damaged;
specifically in the thalamus, the basal ganglia, the cerebral cortex, the hippocampus and the
cerebellum. Cognitive deficits, such as memory problems, executive dysfunction and
impairment of attention are likely.
This report describes the case of a 15 years old male adolescent. He got half buried under a
roof avalanche leading to a hypoxic encephalopathy with asphyxia by thorax trauma and
aspiration pneumonia basal on both sides. Out of hospital reanimation was executed after
10 minutes. Resulting consequences were a posttraumatic/post-ischemic delir with
normothermia protocol (1 day) and severe neuropsychological deficits such as a persistent
anterograde amnesia. However, CT- and MRI-scans of the brain gave no evidence of HIE,
specifically in the hippocampus. Prognosis was uncertain. After three days of intensive care
followed by one-month early rehabilitation, a 5-months intensive rehabilitation period,
including multidisciplinary therapies, was carried out. Improvements in all cognitive
domains were registered but memory problems persisted. Nevertheless, reintegration into
work and study environment was successful due to the good interdisciplinary collaboration.
This single case report gives important insights into the treatment of traumatic brain
injuries. It raises the question of how valid brain scans are in terms of neuropsychological
deficits. Furthermore, it helps to discuss and improve current knowledge on how to help
children with traumatic brain injuries to integrate back into their usual environment
successfully.
Key words: hypoxic encephalopathy, thorax trauma, cognitive deficits, children/adolescent
Isolated pediatric Neurosarcoidosis presenting as bilateral subacute complete
sensorineural hearing loss
D. Brechbühl (1), N. Schöbi (2), G. Mantokoudis (3), M. Steinlin (1)
Division of Neuropediatrics (1), Infectiology,(2) Department of Pediatrics, and ENT-de-
partment (3) Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland)
Case Report: A 10-year-old boy was reffered for progressive hearing loss over the last 4
months. At examination he showed bilateral subacute complete sensorineural hearing loss,
but was otherwise besides significant fatigue free of signs and symptoms. A brain MRI
showed generalized cerebral and spinal leptomeningeal enhancement with bilateral
enhancement of the III, VII and VIII cranial nerve. There was also enhancement of the
internal auditory canal with postcontrast enhancement of the cochlea and the vestibulum.
The cerebrospinal fluid (CSF) analysis showed mild increase of monocytic cells (7) and
increase of protein- and was consistent with blood brain barrier dysfunction, his CSF
angiotensin converting enzyme (ACE) was elevated at 10.16 U/l (0 to 2 U/l). A combined
dural, leptomeningeal and cortical biopsy established the diagnosis of neurosarcoidosis,
showing epitheloid granuloma.
After treatment 2 weeks of high dose steroids there was significant decrease of leptomenin-
geal enhancement, but persistent hearing loss. CSF-monitoring showed normalization of
both the CSF ACE and the blood brain barrier. Treatment was enforced by infliximab and
the patient underwent cochlear implantation to prevent hearing deprivation.
Discusssion: Isolated Neurosarcoidosis is a rare disease mimicking central nervous system
(CNS) infection, inflammatory disorders and neoplasia. Earlier reported manifestations in a
pediatric population include progressive cranial neuropathy (most commonly the VII, but
also involvement of optic, III and VIII nerve are reported). Additional symptoms might be
seizures and hypothalamic dysfunction. Studies showed poor sensitivity and specificity of
CSF-ACE, remaining leptomingeal/brain biopsy the most reliable approach for diagnosis of
Neurosarcoidosis . Early diagnosis would be important to prevent progressive symptoms.
A cross-sectional and longitudinal natural history study of the Swiss cohort of
LAMA2-related congenital muscular dystrophy
Cornelia Enzmann1,2, Andrea Klein1,2,3
1 Pediatric neurology, University Hospital Bern, Inselspital, Switzerland
2 Pediatric neurology, University Children’s Hospital Basel, UKBB, University of Basel,
Switzerland
3 Pediatric neurology, University Hospital Lausanne, CHUV, Lausanne, Switzerland
We report about a planned cross-sectional and longitudinal natural history study. Congeni-
tal muscular dystrophy due to mutations in the LAMA2 gene, also known as Merosin-nega-
tive muscular dystrophy, is one of the most common congenital muscular dystrophies.
LAMA2 encodes for the protein Laminin α2, which belongs to the extracellular matrix.
Missing Laminin α2 leads to an instable connection between the basement membrane and
the sarcolemma of the muscle cell. The disease is diagnosed on the basis of absent or
reduced merosin in immunohistochemistry staining in muscle biopsy, and/or by genetic
testing. Affected children show hypotonia at birth, delayed motor milestones, severe
muscle weakness and usually never achieve independent walking. To date, no pharmacolo-
gical treatment is available, but there are promising results in basic research and a first
clinical trial with omigapil was already conducted in a small cohort. There is little data about
the exact prevalence and the natural disease course.
In this study we aim to increase the knowledge about the natural history of LAMA2-related
MD, provide data about the prevalence in Switzerland and the phenotypic spectrum.
Additionally we want to evaluate different physiotherapeutic assessment tools, to
document clinically relevant changes in MDC1A.
We will evaluate patients with genetically confirmed MDC1A. Data will be collected/ stored
in the existing Swiss neuromuscular registry. A clinically relevant dataset will be collected
over three years with yearly follow-ups. Descriptive statistical analysis will help to characte-
rize the Swiss cohort of MDC1A-patients.
Cognition, behavior, and quality of life in pediatric cancer survivors
Valerie Siegwart1, Valentin Benzing2, Janine S. Spitzhuettl1, 5, Mirko Schmidt2, Michael
Grotzer3, Maja Steinlin1, Kurt Leibundgut4, Claudia Roebers5, Regula Everts1
1 Division of Neuropediatrics, Development and Rehabilitation, University Children`s
Hospital Bern, Inselspital, 3010 Bern, Switzerland
2 Institute of Sport Science, University of Bern, Switzerland
3 University Children`s Hospital Zurich
4 Division of Pediatric Oncology, University Children`s Hospital Bern, Inselspital, 3010
Bern, Switzerland
5 Department of Psychology, University of Bern, Switzerland
Long-term sequelae of cancer and its treatments put pediatric cancer survivors (PCS) at risk
of cognitive and behavioral difficulties, which likely affect quality of life. The aim of the
study was to describe cognition, focusing on executive functions, psychosocial characte-
ristics and health-related quality of life in PCS (with and without central nervous system
(CNS) involvement) and controls. Executive functions are crucial for learning and cognitive
development and contribute strongly to academic achievement and hence merit particular
attention. The influence of processing speed and demographical and clinical risk factors was
considered. 78 PCS, one year or more post-treatment, and 56 controls (aged 7 - 16 years)
were included. Cognitive functioning, behavior and quality of life were assessed using
standardized tests and questionnaires. Results revealed similar fluid intelligence and
significantly poorer executive functions in PCS than controls, with strongest effects in
working memory. Significantly poorer cognitive performance was observed in PCS with
than without CNS involvement reaching its peak in cognitive flexibility. Processing speed
significantly predicted performance in executive functions and CNS involvement was estab-
lished as a key risk factor. Although quality of life was comparable, behavioral difficulties,
such as peer problems, were more pronounced in PCS than controls. Despite evidence for
good intellectual functioning, PCS need to be supervised closely in clinical protocols as
alterations in executive functions may occur years after cancer and likely affect psychosocial
development.
Risk prediction of fever in neutropenia in children and adolescents under
chemotherapy for cancer in a prospective, multicenter trial
Luana Lavieri, Christa Koenig, Roland A Ammann
Division of Pediatric Hematology/Oncology, Department of Pediatrics, Inselspital, Bern
University Hospital, University of Bern, Bern, Switzerland
Background: Fever in severe chemotherapy-induced neutropenia (FN) is the most frequent
potentially lethal complication for children and adolescents with cancer. Risk prediction of
FN would allow for targeted preventive therapies during chemotherapy. This study aimed to
develop risk prediction rules for FN, and FN with safety relevant events (SRE), defined as
serious medical complications (including death, admission to an intensive care unit, severe
sepsis) or bacteremia.
Methods: This multicenter trial prospectively collected data of pediatric patients between 1
to 17.99 years under chemotherapy for cancer. It was designed to compare different
temperature levels for the definition of fever. Patients characteristics and potential risk
factors for FN were collected. Characteristics related to patients (gender, age group), to
diagnosis (type of malignancy, relapse status, bone marrow involvement) and to therapy
and course of disease (chemotherapy intensity, present central venous access device, time
since diagnosis, prior FN episodes) will be analysed with univariate and multivariate mixed
Poisson regression.
Results: Six centers included 269 pediatric patients and 360 episodes of FN, including 72
with SRE, 56 with bacteremia and 30 with serious medical complications. In univariate
analysis, diagnosis of AML, higher chemotherapy intensity, bone marrow involvement and
shorter time since diagnosis were identified as risk factors for FN and FN with SRE. Further
analysis will be added to these preliminary results.
Conclusion: Established and validated pediatric rules predicting the risk to develop FN
during chemotherapy are important to allow specific prophylactic therapies, such as
prophylactic antimicrobial therapy, granulocyte colony-stimulating factor (G-CSF), or other
procedures to protect patients at high risk to develop FN. This study will develop models to
predict the risk for FN, what will allow more specific treatment of individual patients.
High throughput sequencing of CD34+ selected peripheral blood stem cells in
children with various cancer types undergoing autologous stem cell transplantation
Mutlu Kartal-Kaess1,2, Naomi Azur Porret3, Raphael Joncourt3, Vera Ulrike Bacher3,4,
Thomas Pabst5, Gabriela M Baerlocher2,3, Jochen Roessler1,2, Axel Karow1,2
1 Division of Pediatric Hematology & Oncology, Department of Pediatrics, Inselspital,
University Hospital, University of Bern, Bern, Switzerland
2 Department for BioMedical Research, University of Bern, Bern, Switzerland.
3 Department of Hematology and Central Hematology Laboratory, Inselspital, Bern
University Hospital, University of Bern, Bern, Switzerland.
4 Center of Laboratory Medicine (ZLM)/University Institute of Clinical Chemistry,
Inselspital, Bern University Hospital, Bern, Switzerland
5 Department of Medical Oncology, Inselspital, Bern University Hospital, Bern, Switzer-
land
Clonal haematopoiesis of indeterminate potential (CHIP) is present in patients (pts) with
therapy-related myeloid neoplasms at the time of primary cancer diagnosis and before
treatment. Such clones expand under selective pressure from cytotoxic treatment for
primary cancer and can subsequently give rise to overt myeloid neoplasms. CHIP being an
age-related condition, it remains unclear how in children/adolescents cellular stressors such
as cytotoxic therapy influence the expansion of clones carrying mutations in genes
commonly affected in CHIP. Thus, the incidence of CHIP after chemotherapy is less clear in
the paediatric setting.
We retrospectively analysed our cohort of paediatric pts undergoing autologous stem cell
transplantation following intensive chemotherapy for mostly solid tumours. We aimed to
investigate whether the stress from cytotoxic therapy promotes the expansion of clones
with mutations in “CHIP-genes”.
We applied a sensitive error-corrected ion torrent NGS based approach on CD34+ selected
stem cell samples of thirty-three (n=33) children and adolescents stored after stem cell
harvest. The panel comprised 18 genes so far described to be implicated in CHIP including
ASXL1, ATM, CBL, CHEK2, DNMT3A, GNAS, GNB1, IDH1, IDH2, JAK2, MYD88 (L265P),
PPM1D, SF3B1, SRSF2, STAT3, TET2, TP53, U2AF1.
We found genomic aberrations in 6/33 (18%) pts with a high incidence of variants mainly in
ATM but also in CHEK2 with variant allele frequencies (VAFs) ~50% indicating potentially
germline variants. We found CHIP typical findings in 1/33 pts (3%) with a mutation in TP53
(VAF 18%) potentially reflecting CHIP. This patient’s stem cells were harboring two
additional variants of unknown significance (VUS) in ATM and CHEK2. Thus, germline
variants and their accumulation might predispose to the development of solid tumors and/
or pave the way to clonal haematopoiesis. In conclusion, we did not find CHIP at the
reported high frequencies in adults. Further studies of more paediatric cancer pts are
needed.
Paraneoplastic pruritus in patients with Hodgkin Lymphoma. A report of two
cases.
Lukas R. Meyer-Landolt, Tobias Dantonello, Jochen K. Rössler
University Hospital of Bern, Department of Pediatric Hematology and Oncology UZKJO
Introduction: Pruritus is a common clinical complaint encountered in pediatric practice,
most causes being dermatological, allergic or infectious in nature. A rare but important
cause for pruritus is malignancy. We report two cases of patients with Hodgkin lymphoma,
in whom pruritus was the first hallmark of disease leading to diagnosis.
Case presentation: The first patient was a 17y old male who developed severe pruritus over
several weeks. No other systemic symptoms were noted. Antihistamine and topical therapy
showed no effect. Further evaluation showed a large mediastinal mass. Aprepitant, a
neurokinin 1 receptor agonist showed some alleviation of itching in case 1.
The second patient was a 14y old female who presented with cervical lymph node
enlargement for further evaluation. In retrospect, she had suffered from plantar pruritus for
4 weeks. Imaging showed a large mediastinal mass leading to respiratory compromise.
Biopsy and/or lymph node extirpation led to the diagnosis of Hodgkin lymphoma in all
patients. After commencing chemotherapy according to standard protocol resolution of the
lymphoma led to resolution of the pruritus.
Discussion: Paraneoplastic phenomena are clinical symptoms linked to malignancy but not
directly mediated through the neoplastic cells. Chronic pruritus can well be of paraneopla-
stic origin in the pediatric population, described in haematological malignancies and
especially Hodgkin lymphoma. The mechanisms are poorly understood. Inflammatory
cytokines, especially IL-31 and IL-6 have been associated with pruritus and are closely
related with the pathophysiology of lymphoma.
Malignancy should be included in the differential diagnosis of chronic pruritus encountered
in everyday clinical practice, especially when the symptom does not respond to standard
treatment. Pruritus can be the first hallmark of disease before life-threatening complications
develop.
Aprepitant is a promising treatment to alleviate paraneoplastic itching, the most important
therapy being the treatment of the underlying disease.
Continuous monitoring of health data with a wearable device in pediatric patients
undergoing chemotherapy for cancer – a feasibility pilot study
Christa Koenig, Roland A Ammann, Jochen K Roessler, Eva Brack
Division of Pediatric Hematology/Oncology, Department of Pediatrics, Inselspital, Bern
University Hospital, University of Bern, Bern, Switzerland
Background: Pediatric patients with chemotherapy-induced neutropenia are at great risk to
develop severe infections. Delay of diagnosis and treatment can result in increased
mortality. It has been shown that infections can trigger changes of vital signs, as heart rate
variability, very early in their course and before further clinical symptoms. Continuous
health data monitoring may detect such changes earlier than discrete measurements.
Non-invasive on-skin wearable devices (WD) could therefore serve as additional diagnostic
tools. We aim to assess the feasibility of continuous monitoring of heart rate in pediatric
patients undergoing chemotherapy for cancer using a WD.
Methods: This feasibility pilot study will include twenty patients undergoing chemotherapy
for cancer, aged 1 month to 17.99 years. The WD investigated is the Everion® by Biovotion,
a light device, worn with an elastic band on the upper arm or thigh. The primary outcome is
≥acceptable quality of monitored heart rate, during a cumulative duration of ≥18/24h per
day during ≥7 consecutive days. Secondary outcomes include other vital signs, their
comparison with discrete measurements, acceptability by patients and parents, reasons not
to wear the WD, side effects, effort for the investigators and accuracy of measurements by
skin types and during activity. Start of this study is planed for October 2019.
Conclusion: No study has proven feasibility of continuous monitoring of health data in
pediatric patients undergoing chemotherapy for cancer. It is not known if children tolerate
the WD and if data quality sustains. Neither is it known, if parents and patients can handle a
WD in outpatient settings. Knowledge gained from this study will enable potential future
studies, which may identify patterns predicting imminent fever or infection in the near
future. This may lead to earlier diagnosis in patients at high risk for severe infection and the
distinction from patients at lower risk.
Screening of peptide ligands for targeted drug delivery to rhabdomyosarcoma
cells
Dzhangar Dzhumashev, Andrea Timpanaro, Michele Bernasconi, Jochen Rössler
Department of pediatric Hematology and Oncology, Inselspital, Bern University Hospital,
Department for BioMedical Research (DBMR), University of Bern
Rhabdomyosarcoma (RMS) is the most frequent pediatric soft tissue sarcoma. Surgery and
conventional multimodal therapy are not efficient for patients with recurrence or metasta-
ses. Potential approach to improve current therapies is encapsulation of therapeutic agents
into actively targeted nanoparticles. In this study, we aimed to evaluate peptides as
targeting ligands for nanoparticles. We selected from the literature peptides described to be
ligands for proteins that are overexpressed in RMS. These proteins are Integrins, Neuropilin
1, EGFR, Nucleolin, Transferrin. In addition, RMS-targeting peptides, TmR, RMS-P3, RMS-I
and RMS-II, previously discovered by phage display screening, were included.
Biotinylated peptides were conjugated to streptavidin-coated fluorescent Quantum Dots
(Thermo Fisher). Experiments were performed on RMS cell lines and as control on
myoblasts and fibroblasts. In total, 17 different peptides were evaluated. For microscopic
analysis, 10nM QD-peptide conjugates were incubated for 1h or overnight (12h) with cells
seeded on slides (Ibidi). QD were washed and cells were fixed with 2% paraformaldehyde
(PFA). For flow cytometry analysis, cells were detached with Accutase and incubated with
20nM QD-peptide conjugates for 1h at 37°C in suspension, washed and fixed with 0.5%
PFA. The cell penetrating TAT peptide was used as a positive control.
Preliminary results revealed remarkable intracellular uptake of QD-TmR conjugates by flow
cytometry and fluorescent imaging after 12h incubation in cell culture medium. The uptake
of QD-TmR conjugates was higher than non-conjugated QDs or QDs conjugated with
negative control (CmR peptide). Binding of F3 (Nucleolin-binding) peptide was detected by
flow cytometry after 1h incubation. Internalization was further studied qualitatively by
fluorescent imaging. In conclusion, our data suggests that QDs are a convenient tool to
evaluate nanoparticle targeting potential of peptides binding to cancer-specific receptors.
At least two peptides were shown to be promising for further development of liposomal
drug delivery system for RMS.
Surfaceome profiling of rhabdomyosarcoma for CAR T cell targets
Andrea Timpanaro, Dzhangar Dzhumachev, Michele Bernasconi, Jochen Rössler
Department of pediatric Hematology and Oncology, Inselspital, Bern University Hospital,
Department for BioMedical Research (DBMR), University of Bern
Pediatric rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children
and adolescents. Although overall 5-year survival rates have improved with the combined
use of surgery, radiation therapy and chemotherapy, in patients with metastatic and
aggressive disease little progress has been made and the prognosis remains poor.
Cell surface proteins are ideal targets to develop more effective and less invasive cancer
treatments. A novel attractive therapy targeting surface proteins is represented by Chimeric
Antigen Receptor (CAR) T cells. CARs are fusion proteins composed of an antigen
recognition domain, a spacer, a transmembrane domain and one or more intracellular signa-
ling domains. CARs targeting CD19 have achieved significant clinical success against
leukemias, but surface targets for RMS are scarce and no effective CAR T cells for RMS are
available so far.
The aim of this study is to investigate comprehensively RMS surfaceome to identify novel
targets. The final goal is to create effective CAR T cells directed against selected surface
proteins.
In order to achieve our objectives, RMS surfaceome was investigated in a panel of ten RMS
cell lines. Primary myoblasts and a fibroblast cell line were used as controls. Surface proteins
were isolated by two different protocols and the surfaceome of each cell line explored by
LC-MS. We used a set of predicted cell surface proteins, derived from public databases, to
filter the results and to select the most expressed putative antigens. Among these, we found
CD276, known to be overexpressed on RMS. Anti-CD276 CARs are being established.
To conclude, our validated protocols and bioinformatics enrichment of LC-MS data revealed
candidates exclusively expressed on RMS. CD276 CARs, used as positive control, will help
to improve the design of novel CARs against the selected targets to eradicate RMS.
Challenges of treating osteosarcoma in a 12-years old patient with Sickle Cell
Disease
Blank, Bettina et al
Introduction: Cancer in patients with SCD is very rare and treatment including chemothera-
py extremely challenging. We report on a boy with homozygote SCD (HbSS) with knee pain
diagnosed for high-grade osteosarcoma of the distal femur successfully treated at our
center.
Case report: The 12-years old SCD patient presented progressive left knee pain that lasted
since several months. He had a history of multiple vaso-occlusive crisis treated by hydroxyu-
rea since the age of two years. A work-up including imaging and a biopsy revealed
osteosarcoma of the distal femur with pulmonary metastases. He received chemotherapy
including cisplatin/doxorubicin and HD-MTX. Hydroxyurea was stopped to avoid drug-in-
teractions and additional bone marrow toxicity. Monthly erythrocyte apheresis was
conducted to reduce HbS < 30%. Furthermore, because of HD-MTX, we stopped
supplemental prophylactic folic acid for SCD. Delayed clearance of MTX occurred four times
after HD-MTX courses and after the 2nd course, the patient developed self-limiting trouble
of conscience suspicious for seizure. We performed femur amputation due to N. peroneus
involvement with free transfer of the left distal tibia, fibula and calcaneus to the proximal
femur. Imaging and histology revealed a poor treatment response and pulmonary
metastases showed progression. In consequence, we added the immunomodulatory drug
mifamurtide to chemotherapy and pulmonary metastases were resected by bilateral
thoracotomy. One year after diagnosis, he is in complete remission and monthly apheresis is
still ongoing.
Discussion & Conclusion: This case highlights several challenges for treating a SCD patient
with osteosarcoma:
1.) diagnosis might be delayed due to skeletal pain mistaken for pain crisis and patients
coping with chronical pain
2.) regular erythrocyte apheresis can prevent sickle cell crisis during treatment.
Gastrin releasing peptide receptor is overexpressed in Wilms tumour patients and
cell lines
Elizaveta Fasler-Kan, Sabrina Ruggiero, Milan Milošević, Dietmar Cholewa and
Steffen M. Berger
Department of Paediatric Surgery, Children’s University Hospital, Bern
Gastrin.releasing peptide (GRP) regulates numerous functions of the gastrointestinal and
central nervous system, including release of gastrointestinal hormones, smooth muscle cell
contraction and epithelial cell proliferation. It is a potent mitogen for neoplastic tissues. The
effects of GRP are mediated through the gastrin-releasing peptide receptor (GRPR). This
receptor is glycosylated and belongs to the family of 7-transmembrane G-protein coupled
receptors that activate the phospholipase C signalling pathway. Recent studies have shown
that the GRPR is aberrantly expressed in numerous cancers such as those of the lung, colon,
prostate, breast and kidney. Interestingly, the GRPR was present only in tumour tissue, but
not in the normal kidney tissue of patients. Wilms tumour (WT) is one of the most common
paediatric solid tumours, occurring with the incidence of 1 in 10,000 births. WT are
complex childhood neoplasms and are frequently composed of various histological cell
types. With current therapy the prognosis is good. With protocols developed by the
National Wilms Tumor Study Group, the survival rate of children with WT reached 85-90%
by the end of the twentieth century. However, some of WT with a specific aggressive
behaviour have adverse clinical outcome in about 10-15% of the cases. The aim of the
study is to investigate the expression of GRPR in WT patients and in paediatric kidney
tumour cell lines. Our RT-PCR data showed that all analysed paediatric kidney tumour cell
lines (WT- 3 ab, WT-CLS 1. SK-NEP-1, G401) overexpress the GRPR.
Association of lung clearance index with survival in patients with cystic fibrosis
J. M. Kurz1, K. Ramsey1, R. Kraemer2, B. Spycher3, R. Fischer Biner4, P. Latzin1, F. Singer1
1 Division of Respiratory Medicine, Department of Paediatrics, University Hospital of Bern
2 Department of Clinical Research, University of Berne
3 Swiss Pediatric Respiratory Research Group, Institute of Social and Preventive Medicine,
University of Bern
4 Department of Pulmonology, Quartier Bleu, Lindenhofspital - Bern
Background: The lung clearance index (LCI), derived by the multiple-breath washout
(MBW) using nitrogen (N2) as tracer gas, is a sensitive marker to quantify ventilation
inhomogeneity in patients with cystic fibrosis (CF). So far, the association between LCI and
survival has not been investigated.
Aim: Examine the association between LCI and risk of death (mortality) or lung transplanta-
tion (LTX, morbidity) in children with CF.
Methods: Retrospective longitudinal study in a clinical cohort of 147 CF patients aged ≥ 3
years including 1585 LCI measurements between 01.01.1980 and 31.12.2005. First, we
performed cross-sectional analyses at baseline, i.e. when patients entered the study. We
averaged LCI (n = 438) across the first three visits per patient to avoid possible confounding
from indication to measure LCI. Primary outcome was the association of baseline LCI and
the compound outcome (death or LTX) determined in 2018. We fitted logistic regression
models adjusting for age, sex, and date of lung function measurement.
Results: At baseline, patients were mean (SD) 8.7 (3.0) years old and 48% were females.
Follow-up was on average 16.9 (7.6) years. Until 12/2018, 20% (29/147) died or received
LTX. Age and sex distribution was comparable between patients who survived and those
who died or received LTX. The latter group entered the study earlier. The crude odds ratio
(OR, 95%CI) for death or LTX by one LCI unit increase was 1.09 (1.03 to 1.15), p = 0.004,
the adjusted OR was 1.16 (1.08 to 1.25), p < 0.001.
Conclusion: This unique data suggests that elevated LCI is associated with mortality in
school-aged children with CF born 19 to 39 years ago. We acknowledge the limitations of
retrospective analysis of observational, routinely collected health data and historical MBW
measurements. Completion of the database and longitudinal analyses is ongoing.
Improving opportunities for physical activity and participation in inpatient settings
for adolescent cancer patients
Sophie Wist1, Rebecca Näff1, Sonja Lüer2, Eva Brack2, Jochen Rössler2, Ruth Stauffer
Lacorcia1
1 Department of Physiotherapy, Inselspital, Bern University Hospital, University of Bern,
Bern, Switzerland
2 Division of Pediatric Hematology/Oncology, Department of Pediatrics, Inselspital, Bern
University Hospital, University of Bern, Switzerland
Background: Physical activity has shown good results reducing cancer related fatigue and
negative side effects related to chemotherapy that induce a reduction of participation in
daily living and lead to a low quality of life in paediatric cancer patients. Clinical experience
indicates that in particularly adolescent cancer patients reduce their activity level during
intensive chemotherapy. Therefore, an investigation was started to clarify the requirements
and desires of our patients in order to improve their physical activity level.
Methods: Based on literature search, semi-structured interviews were developed and
conducted with four patients, four parents and a multidisciplinary team working with
adolescent cancer patients. The answers were analysed for inhibiting and promoting factors
for physical activity. Combining literature and interview results, recommendations and
potential measures were structured and discussed in an interprofessional setting.
Results: An improvement in the opportunities for physical activity and participation for
adolescent cancer patients meets a need and leads to greater quality of life. The interviewed
persons think that Physiotherapists should take a role in prevention, patient education and
as a motivator. Discussion: Although the investigation included only a small number of
participants, the chosen approach has allowed gathering different aspects for improvement
with relatively few resources. Physiotherapy should be involved earlier in the process and
aim to create an environment beneficial to physical activity.
Conclusion: The first interprofessional new measures to enhance physical activity in
inpatient settings have been recently implemented. The process is to be continuously
evaluated regarding its clinical feasibility.
Systematic differences in analysis software versions hamper infant lung
function testing
Marc-Alexander Oestreich, Florian Wyler, Philipp Latzin, MD, PhD, Kathryn Ramsey, PhD
Pediatric Respiratory Medicine, Inselspital, University Children›s Hospital of Bern, Universi-
ty of Bern, Switzerland
Background: Multiple-breath inert gas washout (MBW) is a non-invasive, feasible, and sen-
sitive technique to assess lung volumes and ventilation inhomogeneity in infancy. Poor
agreement among commercially available setups and software versions currently limits
widespread application as a surveillance tool in early cystic fibrosis (CF) lung disease.
Aim: We investigated differences in algorithms between two versions of an analysis
software and their impact on MBW outcomes functional residual capacity (FRC) and lung
clearance index (LCI).
Methods: Measurements were collected in healthy infants and infants with CF. Lung
function was performed within the first weeks of life and at one year of age. MBW was
performed using 4% sulphur hexafluoride (Exhalyzer® D, Eco Medics AG, Switzerland).
Data were analyzed using WBreath® versions 3.28.0 and 3.51.2 (ndd AG, Switzerland).
Results: Preliminary analysis included data from 14 infants (7 in CF). Using the older
software version (v3.28.0), mean (SD) FRC for healthy infants was 25.8 ml/kg (2.67) and
25.1 ml/kg (2.27) for CF patients. Using the newer version (v3.51.2), mean FRC was
systematically 10% lower (23.2 ml/kg (2.53) for healthy and 22.3 ml/kg (2.56) for CF
infants; p=0.002). In addition, LCI was approximately 14% higher using the newer software
with a mean of difference of 1.03 (0.57; p=0.003) for healthy infants and 0.97 (0.42;
p=0.0009) for CF infants.
Conclusion: Different versions of the same analysis software revealed significant and
systematic differences in MBW outcomes. The newer software version (v3.51.2) calculates
FRC and LCI according to recent consensus guidelines and therefore we recommend using
this for future analysis.
Joint effects of prenatal smoke exposure and polymorphisms in the MBL2 gene on
cord blood levels of mannose-binding lectin
A. Soti1, O. Gorlanova2, L. Müller1, J. Usemann2, L.J Schlapbach1,3, M. Kabesch4, U. Frey2,
P. Latzin1, O. Fuchs1
1 Department of Paediatrics, Inselspital, University of Bern, Bern, Switzerland
2 University Children’s Hospital (UKBB), University of Basel, Basel, Switzerland
3 Paediatric Critical Care Research Group, Mater Children`s Hospital, Brisbane, Australia
4 Department of Paediatric Pulmonology, Allergy and Neonatology, Hannover Medical
School, Hannover, Germany
Background: Prenatal smoke exposure is associated with increased risk for respiratory
disease in offspring and impacts on the developing immune system. In addition, levels of
mannose-binding lectin (MBL), a soluble pattern-recognizing molecule of innate immunity,
are also related to respiratory morbidity early in life. So far, joint effects of prenatal smoke
exposure and single-nucleotide polymorphisms (SNPs) in the MBL2 gene and secondly of
MBL levels in umbilical cord blood (UCB) are unknown.
Objective: To investigate whether prenatal smoke exposure influences MBL levels in UCB in
concert with SNPs in the MBL2 gene region.
Methods: We measured MBL levels in UCB in 221 study participants of a birth cohort of
unselected, healthy, term-born infants. We assessed prenatal smoke exposure by questi-
onnaire, validated by cotinine levels in the first urine. After genome-wide chip-based
genotyping (Illumina HumanOmniExpress) we assessed the association of MBL levels with
MBL2 SNPs in a genome-wide association study (GWAS). In regression analyses, we
analysed effect modification of SNPs by smoking, adjusting for known and possible
confounders. For a preliminary assessment of gene-environment interaction (GxE), we used
logistic regression of MBL levels dichotomised at 700 ng/ml or 300 ng/ml levels.
Results: Prenatal smoke exposure significantly decreased MBL levels in UCB (p=0.006).
MBL2 SNPs were significantly associated with MBL levels. Except for a subgroup of SNPs
(n=6/24, rs920727, rs2506, rs2083771, rs2099903, rs1800450, rs11003134), smoking
significantly lowered MBL levels in addition to MBL2 SNPs. Preliminary data so far suggests
that such interaction exists for the downstream SNP rs10824787 and for rs3829168 in the
promoter region.
Conclusions: Prenatal smoke exposure seems to lower MBL levels in cord blood indepen-
dently as well as in a joint effect with MBL2 SNPs. Preliminary data furthermore suggests a
link between genetic and environmental effects on respiratory morbidity early in life.
Lack of Aquaporin 9 Expression in iPSC-Derived Hepatocytes Impairs Urea
Secretion
Laemmle A, Borsuk M, Robinson J, Gallagher RC, Nuoffer JM, Häberle J, Willenbring H
Reprogramming of patient-derived skin fibroblasts into induced pluripotent stem cells
(iPSCs) followed by directed differentiation into hepatocytes (iPSC-Heps) allows modeling
of genetic liver diseases in vitro. We aim to develop an iPSC-Hep-based model of the urea
cycle disorder ornithine transcarbamoylase (OTC) deficiency (OTCD) that can be used to
screen for pharmacological chaperones (PCs) to treat OTCD.
We generated iPSCs from fibroblasts of OTCD patients and controls and differentiated
them into iPSC-Heps. We compared urea cycle enzyme expression and ammonia metabo-
lism between patient-derived and normal iPSC-Heps and primary human hepatocytes
(PHH). We included human fetal and adult liver tissue as additional controls.
Differentiation of OTCD patient-derived iPSCs into iPSC-Heps revealed a disease-specific
phenotype with reduced OTC activity and urea secretion. However, urea secretion was also
low in control iPSC-Heps, which expressed all urea cycle enzymes at levels comparable to
PHH, even after ammonia challenge. iPSC-Heps are known to correspond to immature fetal
hepatocytes rather than mature adult hepatocytes. Therefore, we compared gene
expression profiles of human fetal and adult liver tissue to identify differentially expressed
genes that may be responsible for the low urea secretion in iPSC-Heps. We found that fetal
liver tissue was lacking expression of aquaporin 9 (AQP9), which is required for urea
secretion. Consistent with their fetal state of differentiation, AQP9 was absent in all of our
iPSC-Hep lines and its induction led to a significant increase in urea secretion.
These findings show that patient-derived iPSC-Heps replicate characteristic features of
OTCD. A limitation of this model is low urea secretion by iPSC-Heps because of their fetal
state of differentiation. We identify AQP9 as a target for overcoming this restriction and
realize the potential of iPSC-Heps.
Effect of metformin on ACTH receptor activation and downstream signaling
Shaheena Parween1, 2, Christa E Flück1, 2 and Amit V. Pandey1, 2
1 Department of Paediatric Endocrinology, Diabetology and Metabolism University
Children’s Hospital, Bern; 2Department of Biomedical Research (DBMR), University of
Bern, Bern, Switzerland
Background: The peptide hormone adrenocorticotropin (ACTH or Corticotropin) is a major
component of the stress response system in the Hypothalamus-Pituitary-Adrenal (HPA)
axis. Under stress, it is secreted from the anterior pituitary and stimulates cortisol producti-
on from the adrenal cortex. Changes in ACTH production or action are associated with
multiple disease conditions. In clinical situations like Cushing’s disease, ectopic ACTH
syndrome and congenital adrenal hyperplasia, there is excess ACTH production and
blocking the interaction of ACTH at its site of action would be a therapeutic option.
Currently, effective therapy to block the action of ACTH is unavailable. Insulin-sensitizing
treatment, such as metformin, has been used to ameliorate a few reported cases of adrenal
disorders. However, the exact mechanism of how these insulin-sensitizing drugs affect the
HPA axis is not known. Here we test whether an insulin-sensitizing drug, metformin have a
direct effect on the activity of ACTH.
Methods: Cell based in-vitro assays were performed to test the effect of metformin on
ACTH receptor activation and signaling. For assays, OS3 cells transfected with ACTH
receptor and luciferase reporter plasmids were used. Cyclic AMP (cAMP) generation upon
receptor activation was measured by dual luciferase assay (Promega). The potential to shift
the ACTH concentration-response curve (CRC) was evaluated to characterize the inhibitory
activity of metformin on ACTH receptor activation. Detailed characterization was done to
calculate the 50% inhibitory concentration (IC50) by varying concentration of metformin.
Results: Metformin was found to inhibit the activation of the ACTH receptor and downstre-
am signaling associated with ACTH response. Significant inhibition of ACTH induced
receptor activation upon treatment with 10 mM metformin was observed. Metformin
shifted the ACTH CRC towards the right by half log, indicating antagonism.
Conclusion: Treatment of an insulin-sensitizing drug, metformin reduces ACTH induced
receptor activation and signaling. This study could be useful in developing new strategies
for management of hyperandrogenic states especially associated with excess ACTH.
Expanding the phenotype of the p.R63W mutation of HNF4A – a case report
Mara Grassi
Masterstudent medicine, University of Bern
Background: The dominant mutation p.R63W (also known as p.R76W) in the hepatocyte
nuclear factor 4 alpha (HNF4A) leads to congenital hyperinsulinaemic hypoglycaemia (CHI),
macrosomia and Fanconi-type tubulopathy. It was only in 2014 that this mutation-specific
phenotype was recognized as such, and so far, 15 patients have been reported.
Case Report: We present a new case of a girl born in 2005 with severe neonatal hyperinsu-
linism, renal tubular disease and hepatopathy. Despite extensive investigations, the
diagnosis remained elusive for 12 years. A trio-whole exome sequencing finally showed this
specific mutation.
The case confirms the so far homogeneous phenotype of the p.R63W mutation, but in
addition, she presents some novel features such as liver cirrhosis, secondary mitochondrio-
pathy and several abnormal laboratory parameters (including low urate and FGF-23 levels).
Discussion: We will focus on the new features of this extensively investigated and
well-documented case. Hypotheses on the link between these features, as well as
hyperinsulinaemia, macrosomia and the impact of the p.R63W mutation on liver and
kidneys should result in a better understanding of this mutation-specific phenotype as well
as the function of HNF4A in general.
Conclusion: This case report helps us to gain insight into the role of HNF4A during
development and in metabolic regulation. It also underlines the importance of sharing case
histories.
New Mechanistic Insights into Androgen Production
Emanuele Pignatti1,2, Mihaela Zavolan3, Christa E. Flück1,2
1 Department of BioMedical Research, Inselspital, Bern University Hospital, University of
Bern, Bern, Switzerland
2 Division of Pediatric Endocrinology and Diabetology, Department of Pediatrics,
Inselspital, University Hospital of Bern, University of Bern, Bern, Switzerland
3 Biozentrum, University of Basel, 4056 Basel, Switzerland
Background: Androgens are essential for the development of sex features and for reproduc-
tion. Androgens peak around the age of 8 in both boys and girls during adrenarche, the
functional activation of the androgen-producing zona Reticularis in the adrenal glands.
However, what controls androgen production is poorly understood. At the same time, we
have little knowledge of what causes most androgen-related disorders, including Premature
Adrenarche (PA) and Polycystic Ovary Syndrome (PCOS).
Aim and Methods: To shed light on the mechanisms of androgen regulation, we used
established in vitro models of androgen activation and inhibition, based, respectively, on
serum starvation and metformin treatment of the steroidogenic H295R adrenal cell line. We
profiled mRNAs and miRNAs in parallel, to obtain simultaneous information on transcripti-
onal activity and suppression of translation. For the identification of regulatory gene
networks, gene-enrichment and pattern recognition algorithms will be used. Follow-up
experiments will allow the validation of our in vitro findings in human – these experiments
will include mRNA and miRNA profiling of the zona Reticularis from human adrenal
samples.
Relevance: Altogether, our results will define novel mechanisms of androgen regulation and
indicate candidate biomarkers for the preventive identification of PA and PCOS patients.
Level of glycemic control in pediatric patients with type 1 diabetes in Bern: a
cross-sectional study
Tanja Zingg, Michelle Dennig, Grit Sommer, Christa E. Flück
Department of Pediatrics (Division of Pediatric Endocrinology and Diabetology), Inselspi-
tal, Bern University Hospital, University of Bern, Switzerland
Background: Good glycemic control prevents long-term complications of microvascular and
macrovascular diseases in type 1 diabetes (T1DM). We aimed to investigate whether our
patients had A1c values <7.5% as recommended by ISPAD and how therapy modality,
duration of diabetes and pubertal status affected the metabolic control of our patients. We
also set out to compare our quality of care with our results of 2008 and with other
published data.
Methods: In 2017/18, we enrolled all patients with T1DM who were followed by the
outpatient clinic of the University Children’s Hospital Bern over a period of 6 months in an
observational cross-sectional study. Each patient was assessed once during the observatio-
nal period, including demographic and clinical data.
Results: 160 patients participated in the study, 41% (n=82) were boys and 49% (n=72)
were girls. Patients had a mean age (SD) at time of visit of 12.6 (3.5) years (range 2-17
years) and a mean duration (SD) of diabetes of 4.6 (3.6) years (range 1-16 years). Most
patients, 63% (n=100) received functional insulin treatment, 29% (n=47) used insulin
pump and 8% (n=13) injected insulin on multiple times per day (twice-daily/three-dose).
CGM devices were used by 43% (n=68) of patients in their diabetes management. Mean
A1c was 8% and 71% had A1c >7.5%. Compared to results from our hospital from 2008,
A1c was slightly higher (8% vs 7.6%), but more patients had diabetes for >2 years (80% vs
47%). Patients with T1DM duration >2 years had more often A1c levels above 7.5 % than
patients with duration <2 years (p<0.001). A1c values were significantly lower (p<0.05) in
patients using CGM devices (7.8% vs 8%).
Conclusion: The overall glycemic control was poorer 2017/2018 than in our study from
2008. This may be due to the higher percent of patients with diabetes duration >2 years,
thus with more patients out of the remission phase. Patients wearing CGM devices
performed better. Unfortunately, our patients in Bern did not reach the target A1c set by
ISPAD, similar to results of other diabetes centres in Europe and the United States. This
highlights the importance of regular consultations and extended use of CGM.
Trajectory of microbiota maturation in healthy Bern infants – a network approach
L. Cecchini, D. Marchukov, S. Ganal-Vonarburg, C. Sokollik, B. Misselwitz
Background: Intestinal microbiota composition is fundamental to human health and
undergoes critical changes within the first two years of life. Factors probably influencing the
microbiota are the maternal microbiota and the general environment in Switzerland.
However, the development of the intestinal microbiota is incompletely understood. Gaining
knowledge of the trajectory of microbiota maturation is likely key to the understanding of
the pathogenesis of many pathologies in childhood.
Aims: We aim for a deep understanding of the maturation of the healthy infant intestinal
microbiota regarding composition, diversity and metabolic activities. We aim for identifying
parameters affecting microbiota maturation and effects of the microbiota on infant
outcome.
Methods: We will recruit 120 pregnant mothers who will be followed as mother-baby pairs
until 10 years of age. Infants will be followed clinically to determine adequate growth and
development as well as pathology including abdominal pain. Epidemiological parameter
and infant nutrition will be assessed. We will collect biosamples such as stool, maternal milk
and skin swaps.
Species composition and diversity will be assessed by 16S sequencing. Metagenomic
shotgun sequencing and bacterial mRNA analysis will inform about metabolic potential and
metabolic activity of the microbiota. Mass spectrometry will assess the small molecule
content of stool and maternal milk samples. Network analysis will be used to assess the
complex relationships between bacteria metabolic activities and small molecular content.
Expected results: We expect an increase in complexity and metabolic potential and activity
with age. Microbiota parameters will differ according to nutrition and might predict infant
outcomes such as growth and abdominal pain. Systematic analysis of sequential maternal
and infant bacteria samples from stool, skin and maternal milk will help characterizing
bacterial transfer from mother to infant.
Conclusion: We propose an observational study of healthy Bern mother baby pairs with
clinical characterization and biosampling. Advanced analysis tools will be used to characte-
rize the microbiota and address mechanistic questions.
Human-induced steroidogenic cells from urine consist a new cell-based model
to study adrenal disorders in a personalised manner
Efstathios Katharopoulos
Background: Congenital adrenal hyperplasia (CAH) is an inherited recessive disorder
characterised by insufficient production of steroids by the adrenal cortex that can also lead
to disorders of sex development (DSDs). CAH is managed by life-long hormone replace-
ment, a suboptimal therapy which fails to mirror the physiological feedback loops of the
hypothalamic-pituitary axis. Recently, various mesoderm-derived cells, including urinary
stem cells (USCs), have been differentiated to adrenocortical-like cells. These innovative
differentiation protocols open the doors to new in vitro disease models and cell-based
treatments (possibly permanent) for CAH, and highlight the huge potential of USCs as
starting material.
Aim: We tried to reproduce and establish in our lab a previous protocol for the isolation of
urinary stem cells (USCs) and their characterisation.
Methods: Fresh urine samples were collected, cells were concentrated and expanded in
vitro using a common culture medium enriched with epinephrine, insulin, EGF, transferrin
and T3. Morphology was microscopically examined and expression of CD44+ was tested in
mRNA level.
Results: Urinary stem cell colonies were successfully (75% success rate) isolated after 2-3
weeks. Cells presented characteristic morphology and expressed the CD44+ mesenchymal
marker. Further experiments for the characterisation of USCs are ongoing.
Conclusions and perspectives: We are in the process of establishing a reproducible
protocol to isolate urinary stem cells in our laboratory. We anticipate that this protocol is
going to foster new research lines in our lab, since USC represent an easily-accessible source
of mesoderm-derived pluripotent cells that can be isolated rapidly and in a cost-effective
way. Future efforts will focus on further characterisation of the isolated USC using FACS
and immunohistochemistry with mesenchymal (CD29, CD166) and pluripotent (SSEA-4)
markers and steroid production.
2019
_10
_23_
MZ
Universitätsklinik für KinderheilkundeForschungssekretariat
Inselspital, Universitätsspital Bern
CH-3010 Bern
Tel.: +41 31 632 94 93