expert / pioneering / connected - … logo retina 2016 is kindly supported by an unrestricted...

EXPERT / PIONEERING / CONNECTED

Retina 2016 is kindly supported by NovartisRetina 2016 is kindly supported by Novartis

ALMERA LOGO

Retina 2016 is kindly supported by an unrestrictededucational grant from Novartis

Also supported byScience Foundation Ireland

Silver Sponsorship

Bronze Sponsorship

Thank you to the following companies who providedadditional support to the Retina 2016 conference.

Retina 2016 is on Twitter at #RetinaDublin@fightngblindnes

Contents

1. Retina 2015 Conference Speakers

2. Welcome to Delegates

3. Call for Research

4. Clinical Trials Workshop

5. Scientific Programme

6. Public Engagement Day Programme

7. Story of the Retina Conference

8. The Fighting Blindness Story

9. Medical and Scientific Advisory Board

10. Fundraising

11. Advocacy

12. Counselling

13. Target 5000

14. Speaker Biographies and Abstracts

15. Public Engagement Day Speakers

16. The Geraldine Duggan Award for Young Investigators

17. Poster List

18. Poster Abstracts

19. Organising Committee

20. Fighting Blindness Contact Information

Delegates will be awarded10 CPD points from the IrishCollege of Ophthalmologists.

ALMERA LOGO



Silver Sponsorship

Bronze Sponsorship



ALMERA LOGO



Silver Sponsorship

Bronze Sponsorship



Retina 2016 is supported by Science Foundation Ireland

ALMERA LOGO



Silver Sponsorship

Bronze Sponsorship



Contents

1. Retina 2015 Conference Speakers

2. Welcome to Delegates

3. Call for Research

4. Clinical Trials Workshop

5. Scientific Programme

6. Public Engagement Day Programme

7. Story of the Retina Conference

8. The Fighting Blindness Story

9. Medical and Scientific Advisory Board

10. Fundraising

11. Advocacy

12. Counselling

13. Target 5000

14. Speaker Biographies and Abstracts

15. Public Engagement Day Speakers

16. The Geraldine Duggan Award for Young Investigators

17. Poster List

18. Poster Abstracts

19. Organising Committee

20. Fighting Blindness Contact Information

Delegates will be awarded10 CPD points from the IrishCollege of Ophthalmologists.

4 Call for Expressions of Interest

5 Welcome

6-7 Retina 2016 Speakers

8 Recently Funded Research

9 Workshop Programme

10-12 ScientificProgramme

14-15 Public Engagement Day Programme

16 Fighting Blindness – Cure. Support. Empower.

17 Retina Conference Story

18-19 Organising Committee

20-21 MedicalandScientificAdvisoryBoard

22-23 Rio to Retina – Limited Sight, Unlimited Vision

24-25 Ways to Support Fighting Blindness

26-27 Resources and Referrals

28-34 Speaker Biographies

35-43 SpeakerAbstracts

44 TheGeraldineDugganAward

45-47 Poster List

48-70 PosterAbstracts

Headline Sponsorship

Thank You to our Retina 2016 Supporters Contents

ALMERA LOGO



Silver Sponsorship

Bronze Sponsorship



5

WelcomeCall for Expressions of Interest

Dear Delegate,

On behalf of Fighting Blindness I am delighted and honoured to welcome you to the 2016 Retina Conference. This is the 17th year of this important event in our calendar. Retina 2016 marksthefirstformeinmytenureasCEO.

Since commencing in March 2016 I have been particularly struck by the enormous reputation that the conference hasbuilt,notonlyamongsttheacademicandscientificcommunities but also, and as importantly, for the engagement and consultation of our patient members.

The fact that over 224,000 people are impacted by sight loss in Ireland means the urgency in advancing towards treatments and cures is imperative. Equally the need for equitableandtransparentaccesstothetherapiesmeansthatFightingBlindness’roleasan advocacy body increases in importance and priority.

Inwelcomingyoutoourconferenceitisopportunetoreflectonthelegacyofachievement by those who have gone before us in championing in Ireland and globally the importance of investment by the State to support the quest for improved treatments and cures. I particularly wish to acknowledge the work done by my predecessor as CEO, AvrilDaly.IndeedwearefortunatethatAvrilcontinuestoadvocatenowwithaglobalbriefasCEOofRetinaInternational.FightingBlindnessisdelightedtohosttheofficesofRetina International and see this as evidence of an on-going commitment to collaborate and to secure best outcomes for people living with sight loss.

This conference could not happen without the goodwill of the contributors whose passion for advancing treatments will be evidenced in the presentations and the consultations you will experience. Furthermore the sponsors for our event are critical to it happening and for this we thank them sincerely.

Finally and most importantly we thank our patient members who have entrusted Fighting Blindness to be their collective voice to ensure advancement of treatment and cures, and equity of access to those therapies.

Thank you for continuing our journey together and let our wish for this conference be that it serves as further evidence that our goals will be achieved in a true spirit of partnership, consultation and collaboration.

Warm regards,Kevin WhelanCEO, Fighting Blindness

Fighting Blindness is dedicated to improving the lives of people in Ireland living with vision impairment and blindness. For more than three decades we have been funding ground-breaking research and are committed to facilitating the development and delivery of preventions, treatments and cures for all those affected by sight loss.

Fighting Blindness is always open to unsolicited expressions of interest in the area of retinal research and invites you to contact us to discuss potential projects further. Potential projects should be clearly aligned with the goals of Fighting Blindness and incorporatemeaningfulpublicandpatientinvolvement.AllexpressionsofinterestarereviewedbyourinternationalMedicalandScientificAdvisoryBoardandfullapplications are internationally peer reviewed.

FightingBlindnessalsoperiodicallylaunchesofficialcallsforapplications.Ifyouwouldliketobeaddedtoalisttoreceivenotificationsofthesecallspleaseemaillaura.brady@fightingblindness.ie

For more information about Fighting Blindness and details of past and current funded projects, please visit www.FightingBlindness.ie.

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Follow Retina 2016 on Twitter at #RetinaDublin

@fightngblindnes

Join the Conversation!

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Retina 2016 SpeakersRetina 2016 Speakers

Dr Michael Redmond National Eye Institute, NIH,USA

Prof Robin Ali University College LondonUK

John Delany Senior Counselling Manager, Fighting Blindness

Dr Shaomei Wang Cedars-Sinai,LosAngeles,USA

Christina Fasser Retina International

Dr Daniel Chung Spark Therapeutics

Prof Andrew Lotery University of Southampton UK

Prof Jane Farrar Trinity College Dublin,

Ireland

Prof Elfride De Baere Ghent University, Belgium

Prof Bart Leroy Ghent University,

Belgium

Prof Austin Roorda University of California,

Berkeley,USA

Peter Ryan Paracyclist and Fighting BlindnessAmbassador

Prof Frans Cremers Radboud University Nijmegen Medical Centre, Netherlands

Dr David Gamm University of Wisconsin,

Madison,USA

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Recently Funded Research

9

Retina 2016 Workshop Programme:Thursday, November 10

Remarkable progress has been made in the quest for treatments for retinal dystrophies and many promising therapies are now at the translational and clinical stages of research. During Retina 2016, Fighting Blindness will hold two topic-focused, multi-stakeholder workshops. Participation in these workshops is by invitation only.

Session 1: Chair: Mr David Keegan, Mater Misericordiae University Hospital

9.00am Opportunities in Stem Cell Research Apanelofpioneeringexpertsinthefieldofregenerativemedicineresearch

and on-going cell therapy clinical trials will discuss future perspectives in stem cell research with a view to highlighting current opportunities and gaps in the area. The goal is to ascertain where Fighting Blindness can capitalise on the skills available in Ireland and through collaboration with international researchers have the greatest impact in the area of retinal stem cells.

Session 2: Chair: Prof Robin Ali, University College London

11.30am Closed Session Fighting Blindness-funded researchers present to International Medical and ScientificAdvisoryBoard.

1.30pm Lunch

Session 3: Chair: Prof Brendan Buckley, ICON Plc

2.30pm Consideration of Regulatory Requirements at all Stages of Research This will be explored by all relevant stakeholders including pharmaceutical

and biotechnology industry, scientists, medical professionals, policy makers and people affected by vision loss. Embedding the patient voice at the heart of the conversation, together they will explore the challenges with a view to expediting the process of moving research from the laboratory to clinical trials, and ultimately to the approval of treatments for patients. This not only impacts on the overall goal of delivering cures to individuals affected by sight loss, but also increases the awareness of scientists to the potential commercialisation of their research..

Session 4: Chair: Prof Robin Ali, University College London

5.30pm Closed Session MedicalandScientificAdvisoryBoardMeeting.

Congratulations to our Recently Funded Researchers

Fighting Blindness is delighted to announce the funding of two new projects co-funded by the Health Research Board through the Medical Research Charities Group co-funding scheme.

Prof James Loughman, Professor of Optometry and Vision Science at Dublin Institute of Technology (DIT), together with Mr Ian Flitcroft, Consultant Ophthalmologist at the Temple StreetChildren’sUniversityHospital,willleadthefirstFightingBlindnessfundedclinicaltrialunderthetitle“PioneeringAdvancesforControlofMyopiainChildren-theSHIELDinitiative”.

Congratulations also to Prof Jane Farrar, Dr Paul Kenna and Dr Matthew Carrigan of TrinityCollegeDublinwhohavesecuredfundingfortheirproject‘ApplicationofNextGeneration Sequencing for the Genetic Characterisation of Irish Retinal Degeneration Patients’whichformspartoftheTarget5000initiative.

Fighting Blindness wishes them and all our researchers every success.

Prof James Loughman, Dublin Institute of Technology (DIT) and Mr Ian Flitcroft, TempleStreetChildren’sUniversityHospital along with the team of researchers involved in the SHIELD initiative

Dr Paul Kenna, Dr Matthew Carrigan and Prof Jane Farrar, Trinity College Dublin.

PROGRAMME

Retina 2016 Scientific Programme:Friday, November 11

7:45am Early Registration and Poster set up

8am– 9am Breakfast with the Prof

Prof Robin Ali: Getting Published - Tips on Writing Papers Prof Joseph Carroll: Networking in Science Prof Bart Leroy: Connecting Clinical Practice with Research

as a Clinician-Scientist Prof Elfride De Baere: Funding Opportunities for Young Scientists

8.30am Registration and Poster set up

Opening Address

9.00am Kevin Whelan, CEO of Fighting Blindness

9.15am Loretto Callaghan, Novartis

Session 1: Chair: Prof John Flannery, University of California, Berkeley, USA

9.30am Prof Jane Farrar, Trinity College Dublin “Targeted Therapeutic Strategies for Inherited Ocular Disorders”

10.00am DrMichaelRedmond,NationalEyeInstitute,NIH,Bethesda,USA “New Insights into the Chemistry and Biology of RPE65”

10.30am Peter Ryan, Paracyclist and Fighting Blindness ambassador

10.45am DrDanielChung,SparkTherapeutics,USA “Preparation for Phase 3 Inherited Retinal Dystrophy Gene Therapy

Trial: Primary Endpoint Selection and Development”

11.00am Coffee Break & Poster viewing

Session 2: Chair: Dr Fionnuala Hickey, Fighting Blindness

11.30am Conor Daly, University College Dublin “ABrain-DerivedNeurotrophicFactorMimeticIsSufficienttoRestoreCone

Photoreceptor Visual Function in an Inherited Blindness Model”

11.40am Dr Louise Porter, University of Liverpool, UK “Investigating a Role for Aberrant Epigenetic Regulation in Genetic Eye

Disease: Approaches in a Rare Disease, Brittle Cornea Syndrome Type 2”

11.50am AndrewSmith,UniversityCollegeDublin “IdentificationandCharacterisationofConePhotoreceptorEnrichedFactorsin

ZebrafishUsingCRISPRCas9”

12:00pm Prof Frans Cremers, Radboud University Nijmegen Medical Centre, Netherlands “SolvingtheUnsolvedInheritedRetinalDystrophiesThroughRNAAnalysisof

Patient-Derived Retinal Organoids”

12:30pm ProfAndrewLotery,UniversityofSouthampton,UK “UpdateonSorsbyFundusDystrophy–FromBenchtoBedside”

1.00pm Lunch & Poster viewing

Session 3: Chair: Prof Frank Barry, REMEDI, National University of Ireland, Galway

This session is kindly sponsored by Science Foundation Ireland

2.00pm DrDavidGamm,UniversityofWisconsin-Madison,USA “Advantages and Drawbacks of 3D Retinal Organoid Technology in the

Development of Therapeutics for Retinal Degenerative Diseases”

2.30pm ProfRobinAli,UniversityCollegeLondon,USA “Differentiation and Transplantation of Embryonic Stem Cell-Derived Cone

Photoreceptors”

3.00pm Prof Elfride De Baere, Ghent University, Belgium “Non-Coding Cis-Acting Defects in Retinal Dystrophies: From Locus

Resequencing to Interpretation”

3.30pm John Delany, Senior Counselling Manager, Fighting Blindness “Thinking About the Patient in Everything We Do”

3.45pm Coffee Break

10 11

PROGRAMME

PROGRAMME

12

PROGRAMME

Session 4: Chair: Prof Alberto Auricchio, Federico II University, Naples, Italy

4.00pm Prof Bart Leroy, Ghent University, Belgium “The Bestrophinopathies as Models for Gene Therapy”

4.30pm DrShaomeiWang,Cedars-Sinai,LosAngeles,USA “iPSC-Derived Neural Progenitors Preserve Vision in an AMD-Like Model”

5.00pm ProfAustinRoorda,UniversityofCalifornia,Berkeley,USA “StructureandFunctionAssessmentsinEyeDiseaseUsingAdaptiveOptics”

5.30pm Presentation of Prizes to Winning Oral and Poster Presentations

BestOralPresentation:GeraldineDugganAward

Best Poster Presentation

5.45pm Closing Comments Christina Fasser, President, Retina International

Retina 2016 Scientific Programme:Friday, November 11

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Retina 2016 Public Engagement Day Programme:Saturday, November 12

10.00am Registration Opening Address

10.30am Kevin Whelan, CEO of Fighting Blindness

10.45am Jason Smyth and Orla Comerford, Paralympic Sprinters “LimitedSight,UnlimitedVision”

Session 1: Chair: Prof Tom Cotter, University College Cork

11.00am DrDavidGamm,UniversityofWisconsin–Madison,USA “Takingthefirststepstowardastemcell-basedtherapy:Whatkeeps researchers awake at night (besides everything)?”

11.20am Dr Daniel Chung, Spark Therapeutics “Developing Patient-Relevant Outcomes in Gene Therapy Clinical Trials” 11.40am ProfJohnFlannery,UniversityofCalifornia,Berkeley,USA “An Overview of Current Vision Research” 12.00pm Condition-SpecificBreakoutSessions 1: Retinitis Pigmentosa (RP), Choroideremia, Leber Congenital Amaurosis(LCA)

2: Stargardt Disease, Cone Rod Dystrophy, Leber Hereditary Optic Neuropathy (LHON)

3: Usher Syndrome 4: Age-relatedMacularDegeneration(AMD),DiabeticRetinopathy, Retinal Dystrophy, Retinal Detachment, Retinal Hole

1.15pm Lunch

Session 2: Chair: Dr Breandán Kennedy, University College Dublin

2.15pm DrSallyAnnLynch,TempleStreetChildren’sHospital “What’sinagenome?”

2.30pm KatrinaConnolly,MIACP-Psychotherapist “The importance of self-care”

2.45pm Caitriona Dunne, Fighting Blindness “Advocacy–PowerinNumbers”

Session 3: Chair: Mr David Keegan, Mater Misericordiae University Hospital, Dublin

3.00pm PanelDiscussion:‘PatientandPublicInvolvementinResearchandthe ImportanceofPatientReportedOutcomes’ Emily Liu, Spark Therapeutics Dr Paul Kenna, Royal Victoria Eye and Ear Hospital Christina Fasser, President of Retina International Ms Giuliana Silvestri, Belfast Health and Social Care Trust

3.45pm Christina Fasser, President Retina International

3:55pm Dr Fionnuala Hickey, Fighting Blindness Research Manager

4.00pm Visionaries Choir Performance

14 15

PROGRAMME

PROGRAMME

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Retina Conference StoryFighting Blindness: Cure. Support. Empower.

Fighting Blindness is an Irish patient-led charity funding and enabling world-leading research into treatments and cures for blindness. The organisation was set up as a support group in 1983 by families affected by sight loss and since then has invested over €16.8 million in more than 90 research projects in Ireland.

We provide a professional counselling service to support people and families affected by sight loss. Through education and advocacy Fighting Blindness works to empower everyone in Ireland living with severe vision impairment.

Fighting Blindness is involved with rare, genetic, age-related and degenerative conditions and represents the 224,000 adults and children in Ireland who are affected by severe vision impairment.

Our vision is to cure blindness, support people living with sight loss and empower patients.

The Retina Conference is an annual event organised by Irish patient-led charity Fighting Blindness. 2016 marks the 17th year of the conference. What began as the ‘All-IrelandRetinalResearchersNetwork(AIRRN)”meetingin2000hasgrownfromstrength to strength over the years and is now a three day internationally renowned conference.Theaimoftheconferenceistobringtogethereminentfiguresintheglobalefforttofindtreatmentsandcuresforconditionscausingsightloss.Theeventfacilitates networking between Irish and international colleagues, leading to future collaborations and catalysing the next generation of vision research.

In 2011 the Retina conference was expanded to include a Public Engagement Day. People and families affected by sight loss and members of the public were invited to attend and learn about developments in research and the importance of patient advocacy.Nowinitsfifthyear,thePublicEngagementDayofferspeopleauniqueopportunity to engage with leading clinicians and researchers. It provides a platform forIrishpatientstolearnabouttheirconditionandtheresearchthatisaimingtofindtherapies to treat it.

In 2013 a third dimension was added to the conference in the form of a clinical trials roundtable meeting, with a focus on diseases of the retina. The purpose of this workshop is to discuss some of the major roadblocks that face the development of clinicaltrialsandultimatelyinfluencetheirsuccess.

CureWe aim to promote and facilitate the development of treatments and cures which are accessible to all patients affected by sight loss.

SupportWe are actively developing our counselling service into a nationwide programme, ensuring access and support of the highest standard is available to patients and family members who are living with sight loss.

EmpowerWe work in partnership with all stakeholder groups in the areas of health, science, industry and government to empower patients and to achieve the greatest impact in the global fightagainstblindness.

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Retina 2016 Organising CommitteeRetina 2016 Organising Committee

Dr David Keegan Consultant Ophthalmic Surgeon, Institute of Ophthalmology, Mater Misericordiae University Hospital, Dublin

Kevin Whelan CEO, Fighting Blindness

Dr Laura Brady ResearchOfficer,Fighting Blindness

Anna Moran Head of Operations,Fighting Blindness

Prof Tom Cotter Professor of Biochemistry, University College Cork

Prof Alan Stitt Centre Director, School of Medicine, Dentistry and Biomedical Science, Queen’sUniversityBelfast

Dr Breandán Kennedy AssociateProfessor,

School of Biomolecular and Biomedical Sciences, University College Dublin

Dr Fionnuala Hickey Research Manager,

Fighting Blindness

Caitríona Dunne CommunicationsandAdvocacy

Executive, Fighting Blindness

Mari McCafferty Finance Manager,Fighting Blindness

Dr Paul Kenna Clinical Ophthalmologist, Royal

Victoria Eye and Ear Hospital, Dublin and Senior Clinical

Research Fellow, Ocular Genetics Unit at Trinity College Dublin

Dr Giuliana Silvestri Consultant Ophthalmic

Surgeon, Belfast Health and Social Care Trust, and Honorary Senior Lecturer, Queens University Belfast

Chair

Chair

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Medical and Scientific Advisory Board Medical and Scientific Advisory Board

FightingBlindnesshasastronganddynamicMedicalandScientificAdvisoryBoard(MSAB).TheMSABplaysavaluableroleinadvisingtheBoardofDirectorsandits Research Sub-Committee, assisting in appraising new grant applications and identifying international peer reviewers. It also monitors on-going programmes and where appropriate, offers recommendations regarding research policies and strategic plans, helping the executive to ensure that the excellent quality of our research is maintained.

TheMSABincludesIrishandinternationalrepresentatives,allofwhomareexpertsin medical research, including but not limited to, retinal research. The group is chairedbyProfRobinAliwhoservesastheChiefScientificOfficerforFightingBlindness.

Prof John Flannery Professor of Neurobiology, University of California, Berkeley,USA

Prof Frank Barry Professor of Cellular Therapy, Regenerative Medicine Institute (REMEDI), National University of Ireland, Galway

Prof Alberto Auricchio Professor of Medical Genetics, Frederico II University / TIGEM, Naples, Italy

Prof Brendan Buckley ChiefMedicalOfficer,ICONPlc.AndClinicalProfessor,

Pharmacology and Medicine, University College Cork

Prof Alan Stitt Director, Centre for Vision

and Vascular Research, Queen’sUniversityBelfast,

Northern Ireland

Prof Joseph Carroll Richard O. Schultz, MD / Ruth Works Professor of Ophthalmology and Professor of Biophysics and Cell Biology, Neurobiology&AnatomyattheMedical College of Wisconsin.

Prof Robin Ali Professor of Human

Molecular Genetics, Institute of Ophthalmology, University

College London

22 23

Rio to Retina – Limited Sight, Unlimited Vision Rio to Retina – Limited Sight, Unlimited Vision

Rio to Retina – Limited Sight, Unlimited Vision

We’redelightedtohavethreemembersofTeamIrelandspeakatRetina2016,fresh from the Paralympic Games in Rio de Janeiro in September! Peter Ryan, Orla Comerford and Jason Smyth are fantastic friends of Fighting Blindness and wonderful ambassadors for our community.

PeterRyanwillspeakaspartoftheScientificProgrammeonFriday,November11.Rio2016wasPeter’sfirstParalympicGames,buthehaspreviouslyrepresentedIrelandat both the Road and Track World Championships. Peter was diagnosed with Leber hereditary optic neuropathy in 2010. He will be speaking about coming to terms with his diagnosis and living with vision impairment, as well as his journey to the Paralympic Games.

Orla Comerford and Jason Smyth, who both have Stargardt disease, will speak at the Public Engagement Day on Saturday, November 12. Orla Comerford took part in her firstParalympicGamesinRio,competinginthe100mintheT13category.OrlarecentlycompletedherLeavingCertandbeganstudyingattheNationalCollegeforArtandDesign, so it has been a busy year for her.

YoumayrememberJasonSmythfromlastyear’sRetinaConference.Jasoncompetedin his third Paralympics this year and once again he took the games by storm, winning the100mintheT13category.ThisisJason’sfifthParalympicgoldmedal,havingwonboth the 100m and 200m in the Beijing 2008 and London 2012 Paralympic Games. Nicknamedthe‘UsainBoltofParalympics’,JasonholdstheParalympicWorldRecordsforboththe100mand200m.Hewascrowned‘FastestParalympianonthePlanet’lastyear at an event in Rio to celebrate one year to the Paralympic Games.

Jason and Orla will speak about their experiences of the Paralympic Games in Rio and offersomeperspectiveonthedifferencebetweencompetinginyourfirstGamesandyour third.

Thank you to Peter, Orla and Jason for their contribution to Retina 2016, and to the work of Fighting Blindness.

Pictured opposite from top: Peter Ryan, Orla Comerford and Jason Smyth.

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Ways to support Fighting Blindness Ways to support Fighting Blindness

CHALLENGE YOURSELFJoin us at Hell & Back, Runamuck, Glendalough Trail, Mini Marathon – we have lots of fun events and challenges to choose from!

DENIM-STRATE YOU CARERock out in denims for the day and organise a Jeans for Genes fundraiser at work to help raise funds and awareness for people affected by genetic blindness and visual impairment.

CHARITY OF THE YEARNominateFightingBlindnessasyourorganisation’sCharityoftheYear.Wewill work together to develop a tailored corporate engagement plan best suited to you and your team.

TEAM BUILDINGConsider teaming up with Fighting Blindness to organise a Blindfold Challenge or Dine in the Dark team building event for you and your colleagues.

FESTIVE FUNWith silly season just around the corner it is time to pick up some Christmas cards, available today at the Fundraising Hub. Or why not ho-ho-hold a Christmas Jumper Day intheofficenextmonthforFightingBlindness?

For more information or other ways to get involved please visit www.FightingBlindness.ie/getinvolved, contact [email protected] -oryoucanpopovertotheFundraisingHubtodaywhereourfundraisersAmandaandEmma will be happy to help you and answer any questions.

Clockwise from top left: • The Googlers enjoying their Blindfold Challenge team building day. • ICON plc Firecrest denim-strate they care by supporting Jeans for Genes • Mater Private Hospital wore a pair to show they care about Jeans for Genes • Super supporters brave the Runamuck Challenge for Fighting Blindness

Go the extra mileFighting Blindness is 90% funded through fundraising. That is why we rely so much on the kindness and generosity of people like you in the community, and of the organisations you work with.

There are so many ways you can get involved:

Resources and ReferralsResources and Referrals

26 27

Inherited retinal diseases represent some of the most genetically complex of all rare diseases, involving over 250 genes. Until now, one of the challenges confronted by someone with such an inherited retinopathy has been the lack of a precise diagnosis. Recentadvancesingene-specificandotherpromisingtherapiesholdthepotentialtoimprove the lives of these people in Ireland. This makes the need for genetic diagnosis all the more pressing as many of these therapies cannot be administrated without precise knowledge of the underlying mutation to be treated.

Recognising this fact, Fighting Blindness launched Target 5000 in 2012. This programme aims to identify the disease causing gene mutation in the 5,000 people estimated to be living in Ireland with an inherited retinal degenerative condition.

This all-Ireland programme is led by three ophthalmologists: Dr Paul Kenna in the Royal Victoria Eye and Ear Hospital; Mr David Keegan in the Mater Misericordiae University Hospital and Ms Giuliana Silvestri of the Belfast Health and Social Care Trust. Genetic screeningandidentificationofdiseasecausingmutationisbeingcarriedoutatTrinityCollege Dublin and Baylor College of Medicine, Houston, Texas.

The information obtained from Target 5000 will provide further insight into the nature and inheritance pattern of inherited retinal degenerations. Target 5000 will also incorporate the development of a National Patient Registry.

How to Refer a Patient

We would ask all optometrists and ophthalmologists to support us in achieving this ambitious goal by referring any interested patients.

To do this please contact Laura Brady 01 6789 004 or [email protected].

We will then discuss the project and process with the patient, answer any questions the patient may have and refer them to one of the project ophthalmologists.

Fighting Blindness provides information and support for people and families affected by vision loss. This includes information about conditions, supports available, and educational materials.

You can refer people to Fighting Blindness by giving our contact details or general brochuretopatients,orbysendinguscontactdetails,withthepatient’spermission,sothatwe can get in touch with them.

Insight Counselling Service

Copingwithvisionimpairmentcanbedifficult.Thereareoftenstrongemotionsthatfollowon from a diagnosis, including anxiety, frustration, fear, anger and depression. It can be helpful in such circumstances to talk to a professional counsellor in order to receive support and guidance.

TheInsightCounsellingServiceisbasedintheFightingBlindnessoffice,3rdFloor,7 Ely Place, Dublin 2. It provides individual, couple and family counselling; telephone counselling; weekly technology support group in Dublin; weekly mindfulness group in Dublin; monthly peer support groups in Dublin and Cork.

Visionaries School of the Arts

Visionaries is a Fighting Blindness quality of life initiative based in the Fighting Blindness officeeverySaturday.Theschoolsactivitiesincludeachoir,musiclessons,musicappreciation, creative writing and comedy improvisation.

You can contact the Insight Counselling Service on 01 674 6496 [email protected].

Information and Educational Materials

We have a wide range of publications and educational materials that we frequently distributetopeopleasrequested.Wealsoreferpeopletoconditionspecificinformationonour fully accessible website.

Oneofourmostpopularresourcesis‘AGuidetoConditionsoftheRetina’,whichisavailable in a number of formats. The guide sets out information about the causes, symptoms, any available or potential therapies and research into these conditions. It details how the eye works and explains the tests that may be carried out at an ophthalmologyappointment.Anoverviewofhumangeneticsandexplanationofinheritance patterns is also included, as well as information about the research process and details for Irish and international patient, medical and research organisations.

To request information or materials or to put someone in touch with us, please contact us [email protected].

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SPEAKERBIO

GRAPH

IES

SPEAKERBIO

GRAPH

IES

Dr T. Michael Redmond NationalEyeInstitute,Maryland,USA

Dr Redmond is Chief of the Laboratory of Retinal Cell and Molecular Biology (LRCMB) and Head of the Molecular Mechanism Section, LRCMB, of the National Eye Institute (NEI), part of the US National Institutes of Health (NIH; Bethesda, Maryland). Dr Redmond received his PhD in 1983 from University College, Dublin (UCD), under the supervision of Prof Eamonn Duke (Dept. of Zoology, UCD),

followinggraduatetrainingatProfRosalieCrouch’slabattheMedicalUniversityof South Carolina, Charleston, SC. He received post-doctoral training at LRCMB, NEI, investigating interphotoreceptor retinoid-binding protein (IRBP). Dr Redmond was tenured in 1990 at NEI and began to study the gene and protein RPE65, then newly discovered by him and his group. He and his colleagues demonstrated the essential role of RPE65 in vision, that mutations in the human RPE65 gene cause Leber congenital amaurosis, a severe early onset inherited blinding disease, and, thatRPE65isthecrucialretinolisomeraseenzymeofthevitaminAvisualcycleandessentialforretinavitaminAmetabolism.Healsocollaboratedintheground-breaking effort to treat human RPE65 Leber congenital amaurosis by gene therapy in2008.Inaddition,hisgroupwasthefirsttoidentifyandclonethemammalian

Peter Ryan Paralympian and Fighting Blindness Ambassador

Peter Ryan is a paracyclist from Drombane, Co Tipperary. He recently competed in the Paralympic Games in Rio and has represented Ireland at both the Road and Track World Championships. He holds the current National Time Trial Championship title.

Peter is studying Sports Conditioning at LIT Tipperary. He was heavily involved in competitive sport from a young age, his primary passion being hurling. Peter played senior hurling for his local club Upperchurch-Drombane and was part of the Tipperary minor team in 2008. In June 2010 Peter was diagnosed with Leber hereditary optic neuropathy (LHON), a genetic disease that leads to sudden loss of central vision during young adult life. Over the past number of years Peter has given his time as an ambassador for Fighting Blindness, raising awareness about vision impairment and the work of Fighting Blindness.

Dr Daniel Chung Spark Therapeutics

Dr Chung is the Ophthalmic Lead for Clinical Development at Spark Therapeutics. Prior to joining Spark Therapeutics, he was a senior investigator at the FM Kirby Center for Molecular Ophthalmology at the Scheie Eye Institute at the Perelman School of Medicine of the University of Pennsylvania, working in retinal gene therapy and transfer. Concurrently, he served asthescientificadvisorontheRPE65genetherapystudy

teamattheChildren’sHospitalofPhiladelphia(CHOP).DrChungearnedhismedicaldegree from the New York Institute of Technology College of Osteopathic Medicine andcompletedhisresidencyinAkron,Ohio.Hethencompletedfellowshipsinpaediatric ophthalmology and ocular genetics research at the Cole Eye Institute at the Cleveland Clinic, with additional training in retinal gene therapy at the National Eye Institute/NIH in Bethesda, MD. In his current duties, he continues to be connected to theRPE65genetherapytrial,asSparkTherapeuticsisnowthesponsor.Astheglobalclinical ophthalmic lead, he works in the areas of clinical development and operations, medical affairs, research and development and business development.

beta-caroteneoxygenaseenzyme,acloserelativeofRPE65thatcatalyzesthefirststepinformationofvitaminAfrombeta-carotenebyanimals.Inhiscurrentresearchon visual biochemistry, Dr. Redmond and his co-workers study the complex enzymatic mechanism of RPE65, its physiological role in maintenance of visual sensitivity and photoreceptor viability, and other research on retinal pigment epithelium metabolism.

Prof Jane FarrarSchool of Genetics & Microbiology, Trinity College Dublin

Prof Farrar is Director of Undergraduate Teaching and Learning for the School of Genetics and Microbiology, Trinity College Dublin, is on the Board of the Trinity College Institute ofNeuroscience(TCIN).SheisamemberofthescientificadvisoryboardofFoundationFightingBlindnessUSAandwasa Co-Founder, Director and CSO of Genable Technologies, recently acquired by Spark Therapeutics. Prof Farrar has

overtwodecadesofexperienceinthefieldofinheritedeyedisorders.Herresearchinterests include elucidation of the genetic etiology and molecular pathogenesis of hereditary neurodegenerative diseases of the retina. Therapeutic avenues being investigated for these ocular conditions include gene and cell-based approaches. Keyachievementsincludetheidentificationofsomeofthefirstdiseasegenesimplicated as causative of inherited retinopathies using genetic linkage analysis, and more recently, characterisation of disease-causing mutations using NGS-based methodologies and the development of novel technologies for treatment of inherited oculardisordersemployingAAV-mediatedgenedelivery,amongstotherapproaches.Prof Farrar has been PI or Co-PI on many national and international research awards generatingsignificantresearchfunding.AdditionallyProfessorFarrarisaninventoronaportfolioofgrantedpatentsinmanyjurisdictionstogetherwithanumberoffiledpatent applications.

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Prof Robin AliUniversityCollegeLondon,UK

ProfAliisProfessorofHumanMolecularGeneticsatUCLInstitute of Ophthalmology. He is also the Theme Leader for Gene Therapy at NIHR Biomedical Research Centre for Ophthalmology,MoorfieldsEyeHospitalandDirectoroftheUCLGMP gene therapy vector manufacturing facility. He is a Visiting Professor at The Kellogg Eye Center, University of Michigan andFounderandChiefScientificOfficerofMeiraGTxLtd,a

genetherapycompanywithofficesinNewYorkandLondon.ThemainfocusofRobinAli’sresearchisthedevelopmentofgeneandcelltherapyforthetreatmentofretinaldisorders.RobinAliandmembersofhisteamhavereceivednumerousprizesandawards for their work on developing new treatments for retinal degeneration including thePfizer/ARVOKarlCamrasTranslationalAwardin2010andAlconResearchInstituteAwardin2009.In2007hewaselectedtotheUKAcademyofMedicalSciencesandin2009 appointed Senior Investigator of The UK National Institute of Health Research. He is President-Elect of the European Society of Gene and Cell Therapy and has served on

Prof Frans P. M. CremersRadboundUniversityNijmengenMedicalCentre,Netherlands

Prof Cremers, PhD is Professor of Ophthalmogenetics in the Department of Human Genetics and Donders Institute for Brain, Cognition and Behaviour (Radboud University Medical Center, Nijmegen,The Netherlands). He is the director of the Foundation Fighting Blindness Nijmegen Research Center and an expert in molecular genetics of inherited retinal diseases. Hehaspublishedmorethan270papers(e.g.intheAJHG,

Cell,Nature,NatureGenetics,PNAS,Science)and15bookchapters.Heinitiatedand coordinates national (RD5000) and international (European Retinal Disease Consortium-ERDC;www.ERDC.info)collaborationsintheretinaldiseasefield.Histeamidentifiedworldwidethelargestnumber(30)ofgenesmutatedininheritedretinal diseases. This concerns genes responsible for classic retinal diseases, e.g. choroideremia (CHM/REP1), Norrie disease (NDP), and X-linked RP type 3 (RPGR), and frequently mutated genes associated with genetic heterogeneous retinal diseases,suchasCEP290andCRB1inLCA,andEYSandUSH2A(longisoform)inarRP.InadditionTSPAN12andZNF408wereaddedtotheNorrin-β-cateninpathwayinvolved in familial exudative vitreoretinopathy. Recently, he has focused his research on‘solvingtheunsolved’IRDsbyanalysingRNAabnormalitiesinretinalorganoidsdifferentiated from patient-derived stem cells. Emphasis is put on solving Stargardt disease cases with one or no mutation.

Dr David Gamm UniversityofWisconsin,USA

DrGamm,MD,PhD,isanAssociateProfessorofOphthalmology and Visual Sciences, and the Emmett A.HumbleDistinguishedDirectoroftheMcPhersonEyeResearch Institute at the University of Wisconsin-Madison. He also holds the Sandra Lemke Trout Chair in Eye Research, and is a member of the Waisman Center Stem Cell Research Program and the UW Stem Cell and Regenerative Medicine

Center. Dr Gamm earned his medical and doctoral degrees from the University ofMichigan-AnnArborandcompletedhisresidencyandpediatricophthalmologyfellowshipattheUniversityofWisconsin,beingelectedintotheAmericanSocietyforClinical Investigation. Dr Gamm diagnoses and manages a wide range of pediatric eye and vision disorders, including inherited retinal degenerations. However, the majority of his effort is directed toward basic and translational retinal stem cell research. The aims of his laboratory are to investigate the cellular and molecular events that occur during human retinal differentiation, and to generate cells for use in retinal disease modeling and cell replacement therapies. To meet these goals, his lab developed a 3-dimensional optic vesicle (or retinal organoid) culture system using human ES and iPS cells, which has the capacity to model retinal development and disease, as well as to delineate the genetic “checkpoints” necessary to produce particular retinal cell types. By understanding the behavior of these cell types in vitro and in vivo, they hope to optimize strategies to delay or reverse the effects of blinding disorders such as retinitis pigmentosa and age–related macular degeneration.

Prof Andrew LoteryChairofOphthalmology,UniversityofSouthampton,UK

ProfLotery’s,MDFRCOphth,clinicalspecialtyismedicalretina with major research interests in age-related macular degeneration, retinal degenerations, ophthalmic genetics, gene and stem cell therapies. He is Editor in Chief of the journalEye,andChairoftheMedicalAdvisoryBoardofRPFighting Blindness UK. He has been awarded the Nettleship AwardforResearchbytheRoyalCollegeofOphthalmologists

andwaslistedintheTimesasoneoftheUnitedKingdom’stop100doctors.Heandhis team have performed over 70 clinical trials and published over 170 papers in high impact journals including Nature Genetics, Nature Communications, the Lancet and the New England Journal of Medicine.

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Prof Elfride De BaereGhentUniversity,Belgium

Prof De Baere, MD, PhD, is Full Professor at the Ghent University in Ghent, Belgium, Senior Clinical Investigator at the Research Foundation Flanders (FWO) and Head of the DNALaboratoryattheCenterforMedicalGenetics(CMGG)attheGhentUniversityHospital.ProfDeBaere’sresearchisfocused on elucidating the molecular pathogenesis of inherited eye disorders such as retinal dystrophies. Key achievements

includetheidentificationofnoveldiseasegenesforrecessiveanddominantretinaldystrophies, characterisation of genetic defects including structural variations using NGS-based approaches and more recently, study of the role of the non-coding portion of the genome in retinal dystrophies. She is partner of several national and international consortia such as the European Retinal Disease Consortium (ERDC), EyeTN network (FP7-PEOPLE-2012-ITN), the Belgian Medical Genomics Initiative (BeMGI). She is coordinator of a Hercules type 2 consortium (High-throughput next-generation sequencer) and a concerted research action at Ghent University (PersonalizedfunctionalgenomicsinMendeliandiseases).SheisAssociateEditorofClinical Genetics and serves on the advisory boards of different (inter)national funding bodies and research councils. She is vice-president of the College of Genetics. In 2012 she was awarded the Inbev-Baillet Latour Prize for Clinical Research.

John DelanySenior Counselling Manager, Fighting Blindness

John is a graduate of Dublin City University and the Irish Institute of Counselling and Psychotherapy Studies. He is also anaccreditedmemberoftheIrishAssociationforCounsellingand Psychotherapy. He worked as a psychotherapist in the public health service and private practice prior to joining Fighting Blindness as their Senior Counselling Manager in May 2014. John is currently undertaking research into the

lived experience of people with RP. He is also developing a programme for the use of the arts by vision impaired and blind people in a therapeutic setting.

the advisory boards of a number of funding bodies including the UK Medical Research Council (Neurosciences and Mental Health Board and the Regenerative Medicine ResearchCommittee),ResearchtoPreventBlindnessUSAandFightingBlindnessIreland, as well as the advisory boards of a number of pharmaceutical and biotech companies.RobinAliobtainedhisBSc(1988)andPhDinGenetics(1993)atUniversityCollege London, continued by postdoctoral training at the MRC National Institute for Medical Research and UCL Institute of Ophthalmology. He established his laboratory at UCL Institute of Ophthalmology in 1999.

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Prof Bart Leroy GhentUniversity,Belgium

Prof Leroy, MD, PhD, is Chairman and Head of the Department of Ophthalmology at the Center for Medical Genetics at the Ghent University Hospital, where he has been workingsince2001.HewasappointedasAssociateProfessorof Ophthalmic Genetics and Visual Electrophysiology at the GhentUniversityinOctober2009.SinceAugust2013,ProfLeroy has also held a part-time position as an attending

physician at the Department of Ophthalmology and the Center for Cellular & Molecular TherapeuticsatTheChildren’sHospitalofPhiladelphia,PA,USA.Hisresearchmainlyfocuses on genetic eye disease. In collaboration with the team at the University of PennsylvaniaandtheChildren’sHospitalofPhiladelphia,Bartisinvolvedingenetherapy projects for inherited retinal blindness. He is a member and Board member of several professional organisations, including the European Organisation for Vision & Eye Research, of which he is the current Past-President, the International Society for Genetic Eye Disease & Retinoblastoma, the Société de la Génétique Ophtalmologique FrancophoneandtheAcademiaOphthalmologicaBelgica.

Dr Shaomei Wang Cedars-Sinai,LosAngeles,USA

Dr Wang, MD, PhD. obtained her MD in China and PhD in England,UK.Sheenteredthefieldofretinaldegenerativediseases and repair as a post-doc working with Professor Ray Lund,FRS,oneofthepioneersinthefieldattheInstituteofOphthalmology, UCL, UK. Her main research projects are to (i) understand the underlying pathology of retinitis pigmentosa, age-related macular disease and optic nerve neuropathy; (ii)

develop therapeutic approaches to ameliorate these blinding diseases. Her laboratory has a long history of applying cell-based therapy for retinal degeneration. Her teamhaveexploredtheefficacyofarangeofdifferentcelltypes(i)cellstoreplacedefective retinal pigment epithelial (RPE) cells such as human embryonic stem cell derived –RPE cells, (ii) cells that appear to function by releasing growth factors such asperipheralnerveensheathingcells(Schwanncells)orcellsgeneticallymodifiedto release growth factors and (iv) cells with multiple functions such as stem cells and neural progenitors. Their proof of concept studies have provided the pre-clinical dataforthreeprominentfirstinhumanclinicaltrialsforretinaldegenerationusingeitherhumanembryonicstemcells(AdvancedCellTechnology)orumbilicalcordtissue derived mesenchymal stem cells (Johnson and Johnson) or central nervous system derived neural stem cells (StemCell Inc.). Currently, Dr. Wang is working at Regenerative Medicine Institute at Cedars-Sinai medical center. Their group works closely with Dr. Clive Svendsen, Director, on cell based therapy to treat degenerative retinal diseases. Her group has been awarded with grants from NIH, DOD, CIRM and multiple industry sponsored projects. Currently, they are performing an IND enabling study of using neural progenitor cells to treat retinitis pigmentosa. The outmost goal of their research is to use stem cell and gene editing therapies for treating retinitis pigmentosa, age related macular degeneration and optic nerve neuropathy and understand the mechanism of action of stem cells in rescuing vision.

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Prof Austin RoordaUniversityofCalifornia,Berkeley,USAProf Roorda received his PhD in Vision Science and Physics from the University of Waterloo, Canada in 1996. Since that time, he has been pioneering applications of adaptive optics and ophthalmoscopy, including mapping of the trichromatic cone mosaic while a postdoc at the University of Rochester, designingandbuildingthefirstadaptiveopticsscanninglaserophthalmoscope at the University of Houston, tracking and targeting light delivery to individual cones in the human eye

atUCBerkeley,andbeingpartofthefirstteamtouseAOimagingtomonitorefficacyofatreatmenttoslowretinaldegeneration.SinceJanuary2005,he’sbeenattheUC Berkeley School of Optometry where he is a member of the Vision Science and Bioengineeringgraduateprograms.HeisaFellowoftheOpticalSocietyofAmericaandoftheAssociationforResearchinVisionandOphthalmology.NotableawardsaretheGlennA.FryawardfromtheAmericanAcademyofOptometry(2009)andaJohnS.GuggenheimFellowship(2014)andanAlconResearchInstituteAward(2016).In2015,DrRoordaledoneoftheteamsthatwasawardedthefirstroundoffundingthoughtheNationalEyeInstitute’sAudaciousGoalsInitiativeaimedat‘Restoringvisionthroughregenerationoftheretina’.Histeam’sprojectinvolvesthedevelopmentofanovelsystem to measure function of individual retinal neurons in human eyes using adaptive optics, eye tracking and phase-resolved OCT.

Christina FasserPresident, Retina International

Christina Fasser is president of Retina International and of the Foundation “blindekuh” Switzerland, vice-chair of Proraris Switzerland and a trustee of Foundation Fighting Blindness. She began her career as an advertisement manager for international advertising agencies. From 1978 to 1990, she worked at the Swiss Federal Institute of Technology on various projects designedtoaidthedevelopingworld,especiallyinthefieldofFoodScience.AffectedbyRetinitisPigmentosa,Ms.Fasser

became involved in the RP Society as a founding member and has been its president from 1986 to 2004 and its CEO from 1991 to 2014. In 1992, she was elected president of RetinaInternational(formerlytheInternationalRetinitisPigmentosaAssociation).RetinaInternational has at present over 30 member organizations, in which more than 1,300,000 individuals affected by retinal degenerative diseases are active. These organizations collect annually more than 30 million US dollars to be invested into research. In her capacity as president of Retina International, Ms. Fasser is involved in promoting research on national and international levels and coordinating the efforts made by individual countries. Christina Fasser has been a leading voice in the effort to focus attention on the needforscientificresearchtofindacureortreatmentforretinalblindnessforthepast25 years. Diagnosed with RP when she was thirteen, Christina discovered early in life the reasons for her clumsiness in the dark and her inability to compete in sport. Contrary to the expectations of her parents, Christina was not bothered at the time by this tragic news as it gave an explanation for what she was experiencing. It was later in life, when choosing a profession, that the greater effect of the condition was felt as she was unable to follow her dream career path. Through determination she has been successful in her career and has risen to a highly respected position in the retinal research community. AsPresidentofRetinaInternational,ChristinaworkswiththeManagementCommitteeand CEO to facilitate communication between members, researchers, industry and government, and to represent the member organisations at meetings world-wide.

Prof Jane Farrar, Trinity College DublinTargeted Therapeutic Strategies for Inherited Ocular Disorders

GJaneFarrar,SophiaMillington-Ward,NaomiChadderton,ArpadPalfi,MatthewCarrigan,KillianHanlon,AdrianDoherty,MarianHumphries,SophieKiang,GiulianaSilvestri,DavidKeegan,PeteHumphries&PaulFKenna,

Inherited retinal degenerations (IRDs) represent a group of genetic conditions that individuallyarerelativelyrare,butcumulativelyrepresentsignificantnumbersofindividuals. Some disease processes apparent in these inherited ocular disorders are mirrored in more common forms of eye disease including age related macular degeneration(AMD)affectingapproximately10%ofpeopleover70yearsofage.Significantstrideshavebeenmadeindiagnosingtheseconditionsutilizingnextgenerationsequencing (NGS) technologies, approaches that are currently, and that will to a greater extent in the future, transform disease diagnosis, prognosis and our understanding of disease mechanisms. Furthermore such eye disorders have been at the forefront of the development of innovative gene therapies globally providing data showing the power of such therapeutic strategies. Two decades of background research and development in TrinityCollegeDublininthisfieldwillbepresentedfromlinkagestudiestoNGS-baseddiagnostics, as will the generation of innovative approaches focusing on gene-based strategies, for treatment of these debilitating conditions. The evolution of some of these studies in Trinity College Dublin from basic research, to targeted translational research and into a commercial setting will be outlined. Key drivers enabling such a directed evolution will be detailed, as will the characteristics required nationally to facilitate commercialization ofuniversityleadresearch.Theenormouspotentialofsuchtechnologiestosignificantlyimprove quality of life for many patients with devastating disorders will be highlighted.

Dr Michael Redmond, National Eye Institute, Maryland, USANew Insights into the Chemistry and Biology of RPE65

The visual cycle isomerase RPE65, highly expressed in retinal pigment epithelium (RPE), plays a critically non-redundant role in the biochemistry of vision. Recent gene therapy trials,thoughhopeful,suggestthatcompletecorrectionofanRPE65-deficientvisualcycleinhumansisnotassimpleasdroppinginthecorrectcassette.Anentireprocesshasbeen disrupted in a way that we do not fully appreciate. RPE65 itself is a unique member of the carotenoid oxygenase family that does not oxidatively cleave carotenoid carbon-carbon double bonds like its relatives, but instead has evolved into an all-trans-retinyl ester (RE): 11-cis-retinol isomerase. Implicit in this description are both an isomerase function and an O-alkyl ester cleavage function, suggesting a complex mechanism. To date, the isomerase functionality, supplying 11-cis-retinol, is better characterized, with the ester cleavage aspect less well so. To better understand the latter, we are investigating parallelsbetweenthevisualcycleandlipidmetabolism.WehaveidentifiedtriacsinC,an

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inhibitoroflong-chainfattyacylCoAligases,tobeapotentinhibitorofRPE65.Anotheraspect is the putative palmitoylation of RPE65. We have recently found that it is a dynamic andnotastructural(suchasinrhodopsin)modification.IntermsofRPE65’sroleinthebiology/physiologyofvision,ouraimistodefineminimalchromophorerequirementsforphotoreceptor survival in vivo, especially for cones. This is an important consideration forthesuccessofhumanRPE65genetherapy,asinsufficienttransductionisapotentialculpritforlackoflong-termcorrection.UsingRPE65knockinmousemodels,wefindthat, despite near-normal phenotypes at younger ages, reduced (<10% wildtype) RPE65 expression and catalytic activity in hemizygous P25L knockin (KI)/RPE65 knockout (KO) mice slows down chromophore regeneration over successive cycles, causing mild retinal degeneration in older animals. The increasing phenotypic severity from homozygous to hemizygous KI mice indicates a correlation between RPE65 expression level and retinal integrity.ThisstudymaybeusefulinassessingabeneficialtherapeuticdoseforRPE65gene therapy in human subjects.

Prof Frans Cremers, Radboud University Nijmegen Medical Centre, NetherlandsSolving the Unsolved Inherited Retinal Dystrophies through RNA Analysis of Patient-Derived Retinal Organoids

FransP.M.Cremers,R.Sangermano,S.Roosing,M.I.Khan,A.Garanto,V.Richelle,M.Bauwens,E.deBaere,M.Khan,RobW.J.Collin,S.Albert

Moleculargeneticstudiesinthelast25yearsidentified~140genesimplicatedinnon-syndromic inherited retinal diseases. Whole exome sequencing- and gene panel-based sequenceanalysisofIRDsrevealedtheunderlyingvariantsin~60%ofthecases.Inviewof the relatively small number of novel IRD-genes that have been published in the last fewyears,itcanbeconcludedthatwehaveidentified>90%oftheIRD-associatedgenesintermsof‘mutationaldiseaseload’.Apartfromtechnicaldeficiencies(non-coverage,undetected heterozygous deletions) it seems logical that many of the missing variants reside in the non-coding regions that represent 98% of the human genome. We are focusing our research on two large unsolved autosomal dominant RD families for which weidentified2–5Mbcriticalregions,aswellasautosomalrecessivecasesinwhichoneornoDNAvariantwasfound.Currently,wearestudying100Stargardtdisease(STGD1)casesfromtheNetherlandsandtheUnitedKingdomwithoneornovariantintheABCA4gene. In collaboration with Ms. M. Bauwens and Dr. E. de Baere (Ghent, Belgium), we performedABCA4locussequencingandaretestingdeep-intronicandnon-canonicalsplicesitevariantsfortheireffectonRNAsplicing.Weemploytwomethods.First,weclonesegmentsoftheABCA4genein‘minigenes’thatconsistofaGateway-adaptedvectorcontaininghumanrhodopsinexons3and5thatareflankingthemutantorwild-typeABCA4genomicsegmentsofinterest.Inthiswaywewereabletoestablishtheeffectofmanynon-canonicalsplicevariants,amongwhichthemostfrequentsevereABCA4

variant,c.5461-10T>C,whichresultsinexons39or39/40skipping.Second,westudythe effect of variants in retinal progenitor cells or organoids differentiated from induced pluripotent stem cells derived from STGD1 patients. This clearly showed the exon 39 or 39/40skipping‘invivo’.Likewise,weidentifiedseveralotherpseudo-exoninclusionsduetodeep-intronicvariantsindifferentpartsoftheABCA4gene.Manyofthepseudo-exonsareamenabletoantisenseoligonucleotide-basedblockingattheRNAlevelwhichholdsthepromisethatwecandesignmutation-specifictherapiesinthefuture.

Prof Andrew Lotery, University of Southampton, UKUpdate on Sorsby Fundus Dystrophy – From Bench to Bedside

Sorsby Fundus Dystrophy is an autosomal dominant inherited retinal dystrophy which affects people in middle age. It causes both a peripheral retinal dystrophy which can cause night blindness similar to retinitis pigmentosa and a central retinal disease similar to age related macular degeneration. It is probably considerably underdiagnosed due to a lack of recognition of the disease. Many patients may be misdiagnosed as having an idiopathic retinal disease or as age related macular degeneration. In this talk Prof Lotery will discuss novel insights into the disease and innovations in current research and treatment. This will include his long term experience of treatment of the neovascular form of the disease and how Sorsby fundus dystrophy provides insights for both retinitis pigmentosa and macular degeneration through genetic and induced pluripotent stem cell studies.

Dr David Gamm, University of Wisconsin, USAAdvantages and Drawbacks of 3D Retinal Organoid Technology in the Development of Therapeutics for Retinal Degenerative Diseases

Retinal pigmented epithelium derived from human pluripotent stem cells (hPSCs) has already shown promise for disease modeling, drug screening and testing, and transplantation, due in large part to their favorable culture characteristics. More recently, similar applications for hPSC-derived neural retinal (NR) cultures have begun to be investigated. NR can be isolated and differentiated from hPSCs in the form of 3-dimensional optic vesicle-like structures (OVs; also referred to as retinal organoids), which not only produce all major NR cell classes, but do so in a conserved spatiotemporal manner with the capacity to generate laminated NR tissues. However, numerous hurdles currently exist with hPSC-OV technology that need to be taken into consideration when seeking to employ it in therapeutic development and testing. This talk will highlight the current state of hPSC-OV technology and its anticipated applications to the treatment of inherited retinal degenerative diseases.

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Prof Robin Ali, University College London, UKDifferentiation and Transplantation of Embryonic Stem Cell-Derived Cone Photoreceptors.

Daytime vision is highly dependent on cone photoreceptors and retinal degenerations resultingintheirlossarealeadingcauseofblindness.Apotentialtherapeuticstrategyis the replacement of lost cones by cell transplantation. Over the past few years we have focused on characterising and optimising cone differentiation in retinae derived from cultures of mouse and human embryonic stem cells (ESCs). Similar to in vivo development, a temporal pattern of progenitor marker expression is followed by the differentiationofearlythyroidhormonereceptorβ2-positiveconeprecursorsandatlaterstagesofculture,photoreceptorsexhibitingcone-specificphototransduction-relatedproteins. We also establish that the Notch pathway limits temporal progenitor competence for cone genesis, whilst retinoic acid signalling regulates cone maturation, comparable to their actions in vivo. Importantly, ESC-derived cone cells can be isolated in large numbers and transplanted into the adult mouse eyes.

Prof Elfride De Baere, Ghent University, BelgiumNon-Coding Cis-Acting Defects in Retinal Dystrophies: From Locus Resequencing to Interpretation

Over 200 disease genes have been implicated in retinal dystrophies (RDs), accounting for50%-80%ofcases.AlthoughmostmutationsunderlyingRDsarelocatedincodingregions, part of the missing genetic variation in RD is located in non-coding regions of the genome such as untranslated regions (UTRs), promoters, deep intronic regions influencingcis-actingsplicing,orinmoredistantregulatoryregionssuchasenhancers.Here,severaltypesofcis-actingnon-codingvariationinRD,locus-specificapproachesfortheiridentification,andtoolsfortheirinterpretationarediscussed.AfirstexampleisNMNAT1-associatedLebercongenitalamaurosis(LCA9),characterizedbyatypicalmacular phenotype. In patients with this phenotype and without coding mutations, weidentified5’UTRmutationsatadjacentpositionsc.-70andc.-69,withlossofheterozygosity, decreased expression of the mutant transcript, and with decreased activity ofthemutations.Asecondmodelofnon-codingvariationisautosomalrecessiveStargardtdisease in which cis-acting deep intronic mutations have been described. In 66 typical BelgianStargardtpatientswithonecodingABCA4mutation,targetedresequencingoftheABCA4geneanditscis-regulatoryregionrevealeduniquedeepintronicvariants,aputativeregulatory5’UTRvariantandapreviouslydescribeddeepintronicvariantc.4539+2001G>A(knownasV4)inseveralpatients.Forvariantinterpretation,datawereintegratedwithcis-regulatorydatasetsgeneratedbyATAC-seq,ChIP-seq,4C-seq

andRNA-seqininadulthumanretina.Athirdexampleisnon-codingstructuralvariationdisruptingcis-regulatoryelements.AhomozygousdeletionupstreamofEYSwasfoundinapatientwithRDandcharacterizedbyTargetedLocusAmplification(TLA),revealingtheexact nature and complexity of the structural variation.

In conclusion, cis-acting non-coding variation explains a portion of hidden genetic variation inRD.Despitethewideavailabilityofgenome-wideapproaches,locus-specificstrategiesare useful in models of non-coding variation such as autosomal recessive RD with strong genotype-phenotype correlations or with a mono-allelic mutation in an RD gene. Finally, cis-regulatory datasets generated in human retinal tissues are important for the interpretation of non-coding variants in RDs.

Prof Bart Leroy, Ghent University, BelgiumThe Bestrophinopathies as Models for Gene Therapy

Purpose: To describe the genotypes and phenotypes in patients with bestrophinopathies, and illustrate how gene therapy could work for these conditions. Methods: Best vitelliform macular dystrophy will be compared with autosomal recessive bestrophinopathy and autosomal dominant vitreoretinochoroidopathy, including current knowledge about mechanisms of disease. Results: Phenotypes of BVMD range from asymptomatic, normal fundi in heterozygous BEST1 mutation carriers, to classic macular egg yolk-like lesions. BiallelicBEST1mutationscauseARB,characterizedbyshallowretinaldetachmentsintheposteriorpole,withhyperautofluorescentdepositsinthewatershedzoneandsuperonasalto the optic disc. In addition, there are small, inner retinal, cystoid changes. Rod-cone dystrophy(RCD) develops later. Subacute angle closure glaucoma is frequent. It would be feasibletoconsidergenetherapyforARBandpotentiallyBVMD.ADVIRCpatientsshowa360° peripheral hyperpigmented band, peripapillary staphylomata, RCD and microcornea. UniquemutationsinoneBEST1allelecauseADVIRC.Electro-oculographyisabnormalinallbestrophinopathiesfirst.Conclusions: Bestrophinopathies are a diverse group of conditionscausedbyeithermono-orbiallelicmutationsinBEST1.WhereasADVIRCisverydifferent,BVMDandARBshowconsiderableoverlap.Thissuggeststhatathresholdof bestrophin protein production, blurring the differences between dominant or recessive mechanismsofdisease,maybethemaindeterminantofthephenotype.AbnormalEOGsindicate primary RPE involvement. Gene therapy would be an option.

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Dr Shaomei Wang, Regenerative Medicine Institute, Cedars-Sinai, Los Angeles, USAiPSC-Derived Neural Progenitors Preserve Vision in an AMD-like ModelBinLu,SergeyGirman,YuchunTsai,BenjaminBakondi,MelissaKJones,CliveNSvendsen and Shaomei Wang

Age-relatedmaculardegeneration(AMD)isthemajorcausesofirreversibleblindness.Theimpacttopatients’qualityoflifeandeconomiccostsranksAMDamongthetophealthcareconcerns.AMDisamultifactorialdiseasecausedbytheinteractionofenvironmentalfactors,agingandgeneticpredisposition,thusincreasingthedifficultyforrationaldrugdesign.AMDpathogenesisinvolvesacascadeofdetrimentaleventsbeginningwithRPEandBruch’smembranechangesthatculminateinthedeathofphotoreceptorcellsand adjacent RPE ultimately leading to central vision loss. Unfortunately, no effective treatmentisavailabletoslowdiseaseprogressionformajorityofAMD.Stem/progenitorcell therapies have shown tremendous promise for treating retinal degenerative diseases. Currently,therearethreeclinicaltrialsfordryAMDintheUS(ClinicalTrials.gov)andtwotrialsforwetAMDinJapanandUK.TheclinicalstrategiesforthesetrialsaretouseRPEcells to replace the defective host RPE, or to graft cells that slow down the progression of degeneration. The major questions for RPE replacement are: Do the grafted RPE survive,attachtoBruch’smembraneandworkasRPEcells?BothinvitroandinvivostudiesshowedthatRPEcellshavelimitedabilitytosurviveondisease(AMD)Bruch’smembrane. Combined treatments to improve host degenerative environment and promote RPEsurvivalareneededtorescuevisioninAMDpatients.Further,ourrecentstudydemonstrated that human neural progenitor cells (NPCs)-derived from induced pluripotent stem cells (iNPCs) offer dramatic photoreceptor and vision preservation, survive for long term and migrate in the sub retinal space with self-secreting extra-cellular matrix. These cells can phagocyte photoreceptor outer segments (POS), therefore compensate for defective POS phagocytosis by host RPE cells. iNPCs avoid allograft immunogenicity and ethical concern, and allows for limitless production, represent a promising cell source for future personalized therapies.

Prof Austin Roorda, University of California, Berkeley, USAStructure and Function Assessments in Eye Disease Using AO

Adaptiveopticsimagingandfunctionaltestingonacellularscaleisofferingnewinsightson structure and function in retinal disease. This talk will present on the relationships between foveal cone spacing and visual acuity in patients with retinal degenerations. Prof Roorda will also present results of an interesting observation in some eye diseases wherebyconesdonotappearinconfocalAOSLOorinOCTimages,yetretainfunctionand have - in some instances - recovered full functional and structural properties over time.This‘dysflectivecone’phenotypeiscommontoseveralretinaldiseasesanditsidentificationmaybeimportantinidentifyingandselectingpatientsfortreatmentsandmonitoring progression of retinal disease.

The Medical and Scientific Advisory Board of Fighting Blindness reviewed all abstracts submitted and chose the following three for oral presentation.

A Brain-Derived Neurotrophic Factor Mimetic is Sufficient to Restore Cone Photoreceptor Visual Function in an Inherited Blindness Model. Conor Daly,LisaShine,TheresaA.Heffernan,SudhakarDeeti,AlisonReynolds,JohnJ.O’Connor,EugèneT.Dillon,DavidDuffy,WalterKolch,GerardCagney andBreandánN.Kennedy

UniversityCollegeDublin,Ireland

Controversially,histonedeacetylaseinhibitors(HDACi)areinclinicaltrialsforthetreatment of inherited retinal degenerations. Previous studies report that patients with inheritedrodphotoreceptordegenerationshowimprovedvisualfieldsandacuityfollowingtreatmentwithvalproicacid,anapprovedanti-epilepticandHDACi.Utilisingthezebrafishdye-/-modelofinheritedblindnesswedeterminediftreatmentwithanHDACipanelcanrescue cone photoreceptor-mediated visual function. Visual function was assessed by optokinetic response and visualmotor behavioural response assays. Cone photoreceptor outer segment (OS) morphology, ciliary marginal zone (CMZ) apoptosis and cone photoreceptor outer segment (OS) length were assessed by light microscopy. Larvae weredrugtreatedwith1µMTSA,6µMScriptaid,10µMMC1568,10µMMS275and10µM7,8-dihydroxyflavonehydratefrom3-5dpfat28.5°C.UnbiasedproteomicanalysisoflarvaleyesbyLC-MS/MSandIngenuitypathwayanalysis(IPA)uncoveredHDACialteredsignalling pathways. TrkB signalling was pharmacologically inhibited via co-treatment with ANA-12andBDNF/TrkBsignallingwasanalysedbywesternblot.dye-/-exhibitdefectivevisual behaviour, including impaired optokinetic (OKR) and visualmotor (VMR) responses compared to unaffected siblings. dye-/- display defects in retinal morphology including increased cell death in the ciliary marginal zone (CMZ), decreased length of photoreceptor outersegments(OS)andunevenlydistributedoralignedphotoreceptors.HDACitreatmentresultsinsignificantlyimprovedOKR(~43fold,p<0.001)andVMR(~3fold,p<0.05).HDACitreatmentrescuedgrossmorphologicaldefects,reducedCMZcelldeathby80%and increased photoreceptor OS length. Unbiased proteomic analysis of dye-/- eye extractsuncoveredincreasedBDNF-TrkBandAktsignalingaskeymediatorsofHDACirescue.Inagreement,co-treatmentwithANA-12,aTrkBantagonist,significantlyblockedHDACirescueofvisualfunctionandAktphosphorylation.Notably,soletreatmentwithaBDNFmimetic7,8-dihydroxyflavonehydratesignificantlyrescueddye-/-visualfunction(~58foldincreaseinOKR,p<0.001,~3foldincreaseinVMR,p<0.05).

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Investigating a Role for Aberrant Epigenetic Regulation in Genetic Eye Disease: Approaches in a Rare Disease, Brittle Cornea Syndrome TypeL.F. Porter,G.GGalli,S.Williamson,J.Selley,D.Knight,N.Elcioglu,A.H.Lund, R. Bonshek , F.D. Manson and G.C. Black

UniversityofLiverpool,UnitedKingdom

Introduction: Epigenetics refers to changes in gene expression that do not relate to theDNAsequenceitself,encompassinghistoneandDNAmethylation,chromatinmodificationsandmicroRNAs.Epigeneticchangesarekeytotissue-specificphenotypes.Here we present an approach to uncovering epigenetic dysregulation in Type 2 brittle cornea syndrome (BCS2). BCS2 is an inherited connective tissue disease with a devastating ocular phenotype caused by mutations in the transcription factor PRDM5. PRDM5ishypothesisedtoexertepigeneticeffectsthroughhistoneandDNAmethylation,and a powerful role for PRDM5 in the reprogramming of haematopoetic stem cells has also been shown. Methods and results: First we report abnormal retinal vascular morphologyintheeyesoftwocousinswithBCS2(PRDM5Δexons9-14)usingimmunohistochemistry,andminedatafromskinfibroblastexpressionmicroarraysfrompatientswithPRDM5mutationsp.Arg590*andΔexons9-14,aswellasfromaPRDM5ChIP-sequencing experiment. Gene ontology analysis of dysregulated PRDM5 target genes reveals enrichment for extracellular matrix (ECM) genes supporting vascular integrityanddevelopment.Q-PCR,andChIP-qPCRconfirmupregulationofcriticalmediatorsofECMstabilityinvascularstructures(COL13A1,COL15A1,NTN1,CDH5)inpatientfibroblasts.WeidentifyH3K9di-methylationatthesePRDM5targetgenesinfibroblasts,anddemonstratethattheBCS2mutationp.Arg83Cysdiminishesinteractionof PRDM5 with repressive complexes, including NuRD complex protein CHD4, and the repressive chromatin interactor HP1BP3, by co-immunoprecipitation combined with mass spectrometry. We observe reduced HP1BP3 staining in the retinas of two cousins lacking exons 9-14 by immunohistochemistry, and dysregulated H3K9 di-methylation in skin fibroblastsofthreepatients(p.Arg590*,p.Glu134*andΔexons9-14)bywesternblotting.ThesefindingssuggestthatdefectiveinteractionofPRDM5withrepressivecomplexes,and dysregulation of H3K9 di-methylation, play a role in PRDM5-associated disease.

Identification and Characterisation of Cone Photoreceptor Enriched Factors in Zebrafish using CRISPR Cas9.Andrew Smith,ConorDaly,VijenderChaitankar,AnandSwaroop,BreandánKennedy


The inability of the retina to detect/transmit light-triggered signals is largely responsible for incurable blinding conditions such as age-related macular degeneration, and cone-rod dystrophy, due to dysfunction or death of photoreceptor cells. There remains a lack of understanding of genes involved in cone development, function and survival, hinting attheexistenceofconespecificorconesensitiveprocesses.Thisresearchaimedtoidentifynovelfactorsexpressedinconephotoreceptorsinzebrafishandmouse,andin the macula of humans, using sequencing technologies. Immunohistochemistry was performed on adult primate retina to identify the subcellular localisation of one of these factors, clul1. Furthermore, this research aimed to characterise a selection of these factors inzebrafishusinginsituhybridisation,andgeneknockdownandknockoutstrategiesusingmorpholinoantisenseRNA,andCRISPRCas9technologies,respectively.27novel,evolutionarilyconserved,cone-enrichedgeneswereidentifiedinzebrafishandmice,usingmicroarray,andRNAseqanalysis,respectively.Thesefactorswereconfirmedtobeconserved in human, and enriched in the retina. FISH revealed that the genes clul1 and es1werespecificallyexpressedintheconephotoreceptorsofadult,anddevelopinglarvalzebrafish.Immunohistochemistryrevealedclul1tobespecificallyexpressedintheoutersegments of photoreceptors in adult primate retina. Knockdown of the gene clul1 resulted inastatisticallysignificanteffectonvisualbehaviour,anddidnotresultinanysignificantmorphologicaldifferencestotheretinaofzebrafishlarvae.Conclusivelythisprojecthasidentifiednovelfactorsenrichedinconephotoreceptors,whereknockdownofoneofthesefactorsinzebrafishhadanegativeeffectonvisualfunctionindicatingapotentialroleforclul1invisualfunction.Thisresearchfurtheraimstocharacteriseknockoutzebrafishandmouse models of novel factors to elucidate their roles.

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The Geraldine Duggan Award1. Progesterone is Neuroprotective in the Retina by Attenuating

Microglial-Driven Degeneration Sarah L. Roche, University College Cork, Ireland

2. The Effects of Zinc Supplementation on Autophagy in Primary Human Foetal Retinal Pigment Epithelial Cells

Eszter Emri, Centre for Experimental Medicine, School of Medicine, Dentistry and BiomedicalSciences,Queen’sUniversityBelfast,UnitedKingdom;2Ocularbiologyand Therapeutics, University College London, London.

3. Phenotype Screening Identifies Vitamin D as a Significant Regulator of Ocular Developmental Angiogenesis

Stephanie Merrigan, University College Dublin, Ireland

4. Oral Administration of Hsp90 Inhibitor, 17-DMAG, Retards Retinal Degeneration in Two Models of P23H Rhodopsin Retinits Pigmentosa

Anna-SophiaKiang,InstituteofGenetics,TrinityCollegeDublin,Ireland

5. A Novel Erythropoietin Derived Peptide has Significant Neuroprotective Efficacy in Diabetic Retinopathy

PaulCanning,Queen’sUniversityBelfast,UnitedKingdom

6. Progesterone Analog-Mediated Redox Response Enhances Cell Survival in Stressed 661W Photoreceptor Cells

AnaRuiz-Lopez,UniversityCollegeCork,Ireland

7. The Polycomb Repressor Complex and Smad 3 Constitute a Switch Enhancing Complex that Interprets TGFβ Signaling to Control Cell Fate in Diabetic Microvascular Complications

DarrellAndrews,UniversityCollegeDublin,Ireland

8. RAB-28, Linked to Retinal Degeneration, Associates with the BBSome and Intraflagellar Transport in C. Elegans

S. P. Carter, UCD Conway Institute, Dublin, Ireland

9. NO-Notch: Nitric Oxide/Notch Signalling Cross-Talk in Vascular Development

HenrykStanikowski,Queen’sUniversityBelfast,UnitedKingdom

Geraldine Duggan (pictured below right) was one of the founding members of Fighting Blindness in 1983. She is fondly remembered for her great wit, no-nonsense attitude and her energy and enthusiasm for life. She showed immense dedication to and belief intheworkofFightingBlindness.AteveryAGMshewouldchallengeourresearchersandvisitingguestswiththesamequestion—theoneoneveryparentandpatient’smind,“Yesthatscienceisgreat,butwhenwillwegetacure?”Thisquestionresonateswith us and remains uppermost in our minds, driving us forward every year as we prepare for another Retina conference.

Geraldine Duggan passed away on November 16, 2011. We honour her every NovemberattheRetinaConference,wheretheGeraldineDugganAwardispresentedinhermemory.Theaward,presentedbyGeraldine’sfamily,goestoayounginvestigator deemed to have submitted the best abstract and given the best oral presentation at the meeting.

Thirty-three abstracts were submitted for Retina 2016. Three of these were selected topresentshorttalksaspartoftheScientificProgramme.Postersareavailableforviewing in the coffee area.

Thank you to the Duggan family for their on-going support of Fighting Blindness and the Retina Conference.

Previous winners of the Geraldine Duggan Award

2015: Dr Matthew Carrigan, Trinity College Dublin and Dr Kirk Stephenson, Mater Misericordiae University Hospital, Dublin

2014: Dr Orla Galvin, University College Dublin

2013: Dr Sarah Chambers, Queens University, Belfast

2012: Dr Alison Reynolds, University College Dublin

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10. Effects of Intravitreal Anti-VEGF Therapy on the Vasculature in Ins2Akita Diabetic and WT Non- diabetic Mice

JudithLechner,Queen’sUniversityBelfast,UnitedKingdom

11. The Synthetic Progestin Norgestrel Acts as an Antioxidant in a Model of Retinal Degeneration by Modulating Nrf2 Signalling

AshleighM.Byrne,UniversityCollegeCork,Ireland

12. Developing Novel Anti-Angiogenic Compounds for Ocular Neovascular Disorders: a Structure-Activity Relationship for a Family of Cysteinyl Leukotriene Receptor Antagonists

AlisonL.Reynolds,UniversityCollegeDublin,Ireland

13. Evaluation of Potential Mitochondrial Therapies Using Novel Complex I Assays

DM Maloney, Institute of Genetics, Trinity College Dublin, Ireland

14. A Sustained Release Formulation of Quininib-Hyaluronan Microneedles Inhibits Cysteinyl Leukotriene Induced Retinal Vascular Permeability.

Orla Galvin, Renasci Ltd, Nottingham, United Kingdom

15. A Novel Concept for Sub-RPE Deposit Formation ImreLengyel,Queen’sUniversityBelfast,UnitedKingdom

16. Verification of ERG Isolation of Human Rod Photoreceptor Activity Using Silent Substitution

John Maguire, Bradford School of Optometry & Vision Science, University of Bradford, United Kingdom

17. Phase I Device Trial of a Next Generation “Injectable” Telescopic Lens Implant for End Stage Age Related Macular Degeneration

Kirk Stephenson, Mater Private Hospital, Dublin, Ireland

18. Standardisation of Irish Inherited Retinal Dystrophy Service Kirk Stephenson, Mater Misericordiae University Hospital Dublin, Ireland

19. microRNA Regulatory Circuits in a Mouse Model of Retinitis Pigmentosa

ArpadPalfi,TrinityCollegeDublin,Ireland

20. A Novel Phenotype Due to SCL24A1 Mutation Conor P. G. Malone, Research Foundation, Royal Victoria Eye and Ear Hospital,

Dublin, Ireland

21. A Variable Phenotype Associated with BBS1 Conor P. G. Malone, Royal Victoria Eye and Ear Hospital Research Foundation,

Ireland

22. Investigating a Role for Aberrant Epigenetic Regulation in Genetic Eye Disease: Preliminary Approaches in a Complex Phenotype, Dry AMD

Louise F Porter, University of Liverpool, United Kingdom

23. Locus Resequencing of ABCA4 in STGD1 Patients Uncovers Novel Deep-Intronic Splicing and Putative Cis-Regulatory Mutations

Miriam Bauwens, Ghent University, Belgium

24. Target 5000: Genetic Characterisation of a Cohort of Inherited Retinal Degeneration (IRD) Patients

AdrianDockery,TrinityCollegeDublin,Ireland

25. Analysis of Photoreceptor Pointing Using Quadrant Pupil Detection Salihah Qaysi, University College Dublin, Ireland

26. Assessing Cone Photoreceptor Structure in Patients with Exon 3 Interchange Haplotypes in the OPN1LW/OPN1MW Gene Array

EmilyJ.Patterson,MedicalCollegeofWisconsin,USA

27. Multimodal Retinal Imaging in Dementia Lajos Csincsik, Queen`s University Belfast, United Kingdom

28. Human Retinal Imaging Using Scanning Laser Ophthalmoscope PrinceSunilThomas,AdvancedOpticalImagingGroup,SchoolofPhysics,University

College Dublin, Ireland

29. Baseline Characteristics of Patients with Neovascular Age-related Macular Degeneration from the Real-world LUMINOUS Study: Global vs Irish Results

Jennifer Coppins, Novartis Ireland Limited, Dublin

30. The Use of 3D Animated Video for Learning Sean Kirwan, University College Dublin, Ireland

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1. Progesterone is Neuroprotective in the Retina by Attenuating Microglial-Driven Degeneration

Sarah L. Roche and Thomas G. Cotter UniversityCollegeCork,Ireland

Purpose: To study the role of photoreceptor-microglia crosstalk in retinal degeneration and neuroprotection in the rd10 mouse model of retinitis pigmentosa. Background: We have previously shown that Norgestrel, a progesterone analogue, promotes photoreceptor survival in the rd10 retina through an upregulation of growth factors. The aim of the current study was to investigate in more detail the cellular events leadingtoNorgestrel’sneuroprotectiveeffects.Specifically,wewishedtostudytherole that photoreceptor-microglia crosstalk might play, as evidence is mounting for microglia contributing to disease progression in retinal degeneration. Methods: Dams of postnatal day (P) 10 rd10 pups were given a Norgestrel-supplemented diet (80mg/kg). Upon weaning, pups remained on Norgestrel. Tissue harvested from P15-P50 micewasusedtoassessmicroglialactivationandinflammatoryprocesses.Inorderto analyse the contribution of photoreceptor-microglia crosstalk to degeneration and neuroprotection, primary rd10 microglia were cultured with photoreceptors in vitro following pre-treatment with Norgestrel. Results: In Norgestrel-treated mice, less pro-inflammatorymicrogliawereobservedatatimewhensignificantpreservationofthephotoreceptor layer was present. This implicates changes in microglial responses, in Norgestrel-mediated neuroprotection. By culturing isolated rd10 microglia with photoreceptors in vitro, we show that rd10 microglia cause neuronal cell death. Importantly, Norgestrel can prime photoreceptors against microglial-derived toxicity and consequentially reduce photoreceptor death. These neuroprotective effects involve an upregulation of the chemokine, fractalkine, in photoreceptors, which acts directly on microglia to modulate activity. This work presents Norgestrel as a promising therapeutic for retinal degeneration and describes novel aspects to its mechanism of action as a neuroprotective in the retina. We are grateful to Fighting Blindness Ireland and Science Foundation Ireland for supporting this work.

2. The Effects of Zinc Supplementation on Autophagy in Primary Human Foetal Retinal Pigment Epithelial Cells

Eszter Emri1,2,LajosCsincsik1,2, Xinyi Yang2,JánosKriston-Vizi3,RobinKetteler3 and Imre Lengyel1,2

Centre for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences,Queen’sUniversityBelfast,UnitedKingdom;2OcularBiologyandTherapeutics,UniversityCollegeLondon,London;3BioinformaticsImageCoreatLaboratoryforMolecularCellBiology,MedicalResearchCouncil,UniversityCollegeLondon,London,UnitedKingdom.

Age-relatedmaculardegeneration(AMD)islinkedtooxidativestress,inflammationand decline in autophagy in the retinal pigment epithelium (RPE). These all contribute totheprogressionofAMDespeciallyattheearlystagesthatarecharacterisedbyaccumulation of extracellular proteins, lipids and trace elements rich deposits. It is suggestedthatsupplementationwithzincisbeneficialtoslowtheprogressionofAMDclinically.Inthisstudyweinvestigatedwhetherzincsupplementationcoulddirectlyinfluenceautophagy.Wecharacterisedtheautophagosomeformationin primary human foetal RPE cells in culture using LC3I/II antibody staining and confocal microscopy. Results using different autophagy modulators, such as ChloroquineandBafilomycinwerequantifiedusingquantitativehigh-throughputimage screening before and after zinc supplementation. We found that zinc supplementation had no effect on autophagosome formation at basal conditions. However,itsignificantlydecreasedtheautophagosomenumber,inducedbyChloroquineorBafilomycin(p<0.05).BasedontheseresultsweproposethatoneofthewayszincsupplementationaffectstheprogressionofAMDisbydirectlyaffectingautophagy in RPE cells.

3. Phenotype Screening Identifies Vitamin D as a Significant Regulator of Ocular Developmental Angiogenesis.

Stephanie Merrigan,SallyHampton&BreandánKennedy UniversityCollegeDublin,Ireland Introduction:Abnormalocularvasculaturegrowthunderpinsage-anddiabetes-

relatedblindness.Currenttreatmentshaveefficacyanddeliverylimitations.Ourobjective was to discover novel inhibitors of vasculature development through phenotype-based screening of a bioactive drug library. Method: 472 ICCB library compounds were screened for inhibition of ocular hyaloid vessel (HV) development inTg(fli1:EGFP)zebrafishlarvae.Hitcompoundsweretestedforinhibitionofnon-ocularinter-segmentalvessel(ISV)development.mRNAandmiRNAexpressionstudies in larval eyes were carried out by quantitative RT-PCR. Safety studies evaluatedretinalmorphologybylightmicroscopy.InvitroviabilityinARPE-19cellswasevaluatedbyAlamarBlueassay.ARPE-19cellfactorsecretionstudieswerecarriedoutbyELISA.Results: Hit compound calcitriol and 6 additional vitamin D receptoragonists(VDRAs)significantlyinhibitedoculardevelopmentalangiogenesis.DespitepresenceoftheVDRinlarvaltrunks,VDRAsdidnotinhibitISVdevelopment.Safety studies showed calcitriol-treated larvae to have normal retinal lamination/morphology.miR21,VEGFaaandVEGFabexpressionwassignificantlyupregulatedin calcitriol-treated eyes and miR150 expression was unchanged. VEGF receptors flt1andkdrlarenotsignificantlyupregulatedbutadosedependantincreaseinexpressionwasseenincalcitriol-treatedeyes.InARPE-19cells;VDRAshadno

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effectoncellviabilityandpreliminarydatashowsTNFαinducedIL-8andGM-CSFexpressiontobeattenuatedbyVDRAtreatment.Conclusion:VDRAssignificantlyandspecificallyinhibitocularangiogenesisduringzebrafishdevelopment.Thisanti-angiogenic activity correlates with increased miR21 expression and an up-regulation ofVEGF.Futurestudieswillevaluatetheanti-angiogenicmechanismofVDRAsandvalidate anti-angiogenic activity in mouse models.

4. Oral Administration of Hsp90 Inhibitor, 17-DMAG, Retards Retinal Degeneration in Two Models of P23H Rhodopsin Retinits Pigmentosa

Anna-Sophia Kiang,AnhNguyen,PaulKenna,JamesKeaney,LawrenceTam,JaneFarrar,MarianHumphries,MatthewCampbellandPeteHumphries

Institute of Genetics, Trinity College Dublin, Ireland

Aim: To investigate therapeutic potential, enhanced oral delivery and mechanism of actionofthehsp-90inhibitor,17-DMAG,inretardingretinaldegenerationinmodelsof Pro23His dominant retinitis pigmentosa. Methods:AlbinoheterozygousP23Hline-3 rats (LaVail lab) and heterozygous P23H knockin mice (Palczewski lab) were firstsubretinallyinjectedwithAAV2/9expressingshRNAtargetingthetightjunctionproteinclaudin-5ornon-targetingshRNAunderthecontrolofdoxycylineinorderthat modulation of inner blood retinal barrier permeability be inducible. MRI and LC/MS/MS analyses were used to assess small molecule uptake into retina following inductionofshRNAexpression/downregulationofclaudin-5.Theanimals’drinkingwaterwassupplementedwith6mg/Kg/week17-DMAGin2mg/mldoxycyclinefor3or 5 weeks for the mice and rats respectively after which electroretinography was carriedout,animalssacrificedandretinalsectionsanalysedwhereappropriate.ClonedcDNAsexpressingWTand/orP23Hrhodopsinwereexpressedincellcultureinthepresenceandabsenceof17-DMAGtoinvestigatemolecularmechanismsinvitro.Proteinextractsfromanimalsinjectedwith17-DMAGintravitreallyorIPwereused for similar in vivo analyses. Results: Downregulation of claudin-5 results in enhancedretinaluptakeofgadoliniumand17-DMAGasshownbyMRIandLC/MS/MS analyses respectively. Retinal degeneration was retarded in the P23H-3 rat as evidenced by electroretinography and histopathology. However, results from the P23Hknockincohortswerelessclearlydefinedasitappearsthatthismodeldoesnottoleratesubretinalinjectionswell.Resultsoftheeffectsof17-DMAGonproteinexpression of ER-resident chaperone, BiP (Grp78), and other UPR-related proteins will be presented. Conclusion:Enhanceddeliveryof17-DMAGretardsretinaldegeneration in P23H-line 3 rats via modulation of cellular stress responses.

5. A Novel Erythropoietin Derived Peptide has Significant Neuroprotective Efficacy in Diabetic Retinopathy

Paul Canning,OliviaO’Leary,Lynsey-DawnAllen,JasenkaGuduric-Fuchs,MichaelBrines, Anthony Cerami, and Alan W. Stitt

Queen’sUniversityBelfast,UnitedKingdom

Exogenousdeliveryoferythropoietin(EPO)hasanti-inflammatoryandtissueprotectiveeffectsinmanydiseasestates.ARA290,anovelEPO-derivedpeptide,ameliorates vascular leakage and capillary dropout in diabetic rodent retina. Since diabetic retinopathy exhibits a neurodegenerative component, we assessed the potentialofARA290topreventdeficitsinneuralretinalintegrityandfunction,linkedtoinflammatorypathwaysuppression.C57BL/6Jmicewererendereddiabeticbystreptozotocin injection; whilst control mice received citrate buffer. Diabetic animals receivedintraperitonealinjectionof;vehicle,30,or60µg/kgARA290for12weeks.At12weeks,retinalfunctionwasassessedbyscotopicelectroretinography(ERG).Retinal thickness was assessed by optical coherence tomography (OCT), and compared to baseline pre-treatment values. Post-mortem analysis of retinal integrity and activation of immune cells was conducted alongside expression of a panel of proandanti-inflammatorycytokines.HbA1clevelsacrossdiabetictreatmentgroupsweresimilarandsignificantlyelevatedcomparedtonon-diabeticcontrolanimals(***p<0.0001;One-WayANOVA)andARA290hadnoeffectonthehaematocritinanytreatedgroups.Roda-waveamplitudewassignificantlydecreasedathigherlight levels in vehicle-treated diabetic animals compared to non-diabetic controls (**p<0.01;two-wayANOVA,Bonferronipost-test).Thisdecreaseinroda-waveamplitudewasattenuatedinthe60µg/kgARA290-treateddiabeticanimals.Thedecrease in ON bipolar cell function (b-wave amplitude) observed in vehicle-treated diabeticmicecomparedtonon-diabeticmice,wasalsosignificantlyattenuatedinthe60µg/kgARA290-treateddiabeticmice(***p<0.0001;twowayANOVA).ARA290treatment reduced retinal thinning observed in the vehicle-treated diabetic animals measured at 12 weeks diabetes, with retinal thickness approaching that observed in non-diabeticanimals.Diabetesmediatedincreasesinpro-inflammatoryTNFα,IL-1β,andICAM-1retinalmRNAlevelswerepartiallyattenuatedwithARA290treatment.ARA290isapotentneuro-protectantinthediabeticretina,preventingneuralretinaldegenerationmediatedbyincreasedpro-inflammatorystimuli,whilstpreservingretinal functionality.

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6. Progesterone analog-mediated Redox response enhances cell survival in stressed 661W photoreceptor cells

Ana Ruiz-Lopez, Alice C. Wyse Jackson, Sarah L. Roche, Ashleigh M. Byrne, Jennifer Moloney, Thomas G. Cotter

UniversityCollegeCork,Ireland

Purpose: Retinitis pigmentosa (RP) is a retinal disease caused by the progressive loss of photoreceptor cells. Despite a substantial understanding of the mechanisms leading to cell death, there is currently no treatment for RP. We have previously shown that a progesterone analogue, Norgestrel, is neuroprotective in the retina mediatedbytheneurotrophicfactorbasicfibroblastgrowthfactor(bFGF).Wehavealso shown that bFGF drives reactive oxygen species (ROS) production which enhances survival of stressed photoreceptor cells. The aim of the current study was to study the role of ROS production in survival responses following Norgestrel treatment. Methods: The 661W photoreceptor cell line was used in the experiments performed. Serum deprivation was used to induce stress for the time indicated. Flow cytometry was used to measure ROS levels, while bFGF levels were measured by rtPCR and western blotting. Cell viability and proliferation were analyzed using MTS assay. Results: Norgestrel enhances an early pro-survival burst of ROS up to 1h in the stressed cells. No changes in ROS are seen in the absence of bFGF action througheitherblockingitsreceptor(PGRMC1)orusingsiRNA,suggestingthisburstis mediated by the up-regulation of bFGF at 30 min following serum starvation (SS). This may in part explain its protective effects on the 661W cells. This ROS production appearstocomefromNADPHoxdiase(NOX)familywhichhavebeenshowntobeinvolved in cell survival. Conclusions: The early Norgestrel-mediated response in retinal cells induces bFGF production which increases ROS levels from NOX proteins in order to enhance cell survival. We are grateful to Fighting Blindness Ireland and Science Foundation Ireland for supporting this work.

7. The Polycomb Repressor Complex and Smad 3 Constitute a Switch Enhancing Complex that Interprets TGFβ Signaling to Control Cell Fate in Diabetic Microvascular Complications

Darrell Andrews,LetiziadeChiara,GiorgioOliviero,ArianeWatson,GerardCagney,ColmO’Brien,JohnCrean


TGFβiswidelyacceptedtoplayakeypathogenicroleinmanydiseaseprocesses,including the progression of retinal disease where it stimulates epithelial dedifferentiation or EMT and the deposition of Extracellular Matrix (ECM) proteins suchascollagenandfibronectinleadingtofibrosis.TGFβsignalsthroughdownstream Smad effectors in particular Smad3 to recruit the transcriptional regulators of EMT and as such this signalling network has been established as a

primecandidatefortherapeuticintervention.Howeverdespitesignificanteffortsinthisarea,todatefewstudieshavedemonstratedclinicalefficacyandhumantrialstodelayorreversefibroticprogressionbyblockingTGFβhaveprovenineffective.AtthecoreoftheproblemremainsafundamentalgapinourknowledgeofhowTGFβsignallingconvergesandinteractswiththecell’stranscriptionalmachinerytoregulategene expression during the initiation and progression of retinal disease. Recent studieshaverevealednovelinteractionsbetweencomponentsoftheTGFβsignallingcascade, master transcriptions (including Oct4, Sox2 and Nanog) and the general chromatinmachineryleadingtothehypothesisthatacombinationofDNAsequence,chromatin environment and cellular context coordinately regulate cellular responses toTGFβsignalling.Usingendogenousimmunoprecipitationcoupledtoamassspectrometrybasedapproach,wehaveidentifiedaninteractionbetweenSmad3andEZH2, a component of the polycomb repressive complex 2 which forms a context dependent switch enhancing complex that regulates cell fate both during neuroretinal specificationandTGFβmediatedepithelialdedifferentiationassociated.Wefurthercharacterise this interaction and delineate its role in the repression of E cadherin using luciferase assays and ChIP-PCR to demonstrate binding of the regulatory complex to the E cadherin promotor. We propose that this complex forms a molecular switch that regulates promoter access through epigenetic mechanisms and controls genesilencingwithclearimplicationsforpotentialtargetingofTGFβindisease.

8. RAB-28, Linked to Retinal Degeneration, Associates with the BBSome and Intraflagellar Transport in C. Elegans

S. P. Carter,V.L.Jenson,A.A.W.M.Sanders,J.Kennedy,M.R.Leroux,B.Kennedy,O.E.Blacque

UCDConwayInstitute,Dublin,Ireland

Cilia are sensory and signalling organelles which project from the surfaces of most eukaryotic cells. Cilia serve different sensory roles, depending on cell type and can detect a wide variety of ligands. For example, the photoreceptor outer segment is aciliummodifiedforthedetectionoflight.VarioussmallGTPasesfacilitateciliumformationandfunction,servingasimportantregulatorsofciliaryproteintrafficking,includingintraflagellartransport.AnumberofsmallG-proteinsarealsoassociatedwith human ciliopathy phenotypes such as retinal degeneration. To identify new ciliarytransport-associatedproteins,wescreenedavailableC.elegansRNA-Seqlibrarydatasetsforgeneswithexpressionprofilesmatchingthoseofknownciliarygenes.Fromthisanalysiswefoundthatcone-roddystrophy-associatedRAB-28isexpressedspecificallyinciliatedsensoryneurons,whereitaccumulatesatthepericiliary membrane (PCM) and undergoes bidirectional IFT. This localisation and transport behaviour is dependent on GTP binding and the BBSome IFT cargo-adaptorcomplex,whichrecruitsGTP-boundRAB-28tothePCM.Functional

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analysesusinganullallelerevealedthatC.elegansRAB-28isdispensableforciliumstructure, function and IFT. However, overexpression of a predicted constitutively activeRAB-28variant(GTP-bound)disruptsciliaryB-tubuleformationandsensorybehaviour.Furthermore,overexpressionofactivatedRAB-28causesdramaticenlargement of the amphid sensory pore, formed by a supporting (non-ciliated) glialcell.Insummary,thisworkidentifiesRAB-28asaBBSome-dependentIFTcargo, with cell autonomous and cell non-autonomous functions at the ciliary base. FurtherinvestigationinavertebratemodelsuchaszebrafishmayshedlightonthepathomechanismofRAB28-linkedcone-roddystrophyandothercilia-relatedformsofretinal degeneration.

9. NO-Notch: Nitric Oxide/Notch Signalling Cross-Talk in Vascular Development

H.Stanikowski,N.Poulose,K.Edgar,P.Mullan,D.M.McDonald Queen’sUniversityBelfast,UnitedKingdom,Ireland

Introduction: The Notch signalling pathway is known to regulate the proliferation andspecificationofendothelialtipandstalkcellsresponsibleforvascularmigration.Disrupted regulation of this pathway is a characteristic hallmark of pathological angiogenesis in a variety of conditions from cancers to retinopathies. Inhibition of this pathway results in increased vascular branching and endothelial cell proliferation. We have previously demonstrated that eNOS overexpression increases vascular branch formation in the murine retinal vasculature. eNOS derived nitric oxide is a highly reactive and pleotropic molecule and therefore has the potential to interact with the Notch signalling pathway at various sites. Understanding how eNOS is regulated during vascular development is crucial to unravelling any potential crosstalk between eNOS and Notch. Methods: Intraperitoneal injections of BrdU solution were given to eNOS GFP overexpressing and wild type P5 and P7 mice 3 hours prior tosacrifice.Eyeswereremoved,dissected,stainedforlectintoidentifytheretinalvascular region; and BrdU to quantify endothelial cell proliferation. Retinas were then subsequently imaged by confocal microscopy. BrdU endothelial cell proliferation and vascularbranchpointformationwerequantifiedmanuallyintheretinalvasculatureof eNOS GFP overexpressing mice and compared with the wild-type. BrdU labelled proliferatingendothelialcellswerequantifiedbasedonselectedcriteriaofhavingan elongated morphology and localised within the lectin stained vascular network. Results: Our current work shows that eNOS overexpression in the developing murine retina enhances endothelial cell proliferation and branch point formation. Further work willinvestigatehoweNOSinteractswithNotchregardingendothelialcellspecificationin the developing retina.

10. Effects of Intravitreal Anti-VEGF Therapy on the Vasculature in Ins2Akita Diabetic and WT Non-Diabetic Mice

Judith Lechner,JoseR.Hombrebueno,MeiChen,HepingXu Queen’sUniversityBelfast,UnitedKingdom

Aim:Anti-vascularendothelialgrowthfactor(VEGF)therapyhasbecomeastandardtherapy for the management of diabetic macular oedema. However, VEGF is an important growth factor for retinal neurons and in a previous study we have shown that multiple intravitreal injections of anti-VEGF induced retinal neurodegeneration inIns2(Akita)diabeticmice.InthisstudyweanalysedtheretinalvasculatureinIns2(Akita)diabeticmicefollowingmultipleintravitrealinjectionsofanti-VEGF.Materials and Methods:Ins2(Akita)diabeticmicewereinjectedintravitreallywith1µlof anti-VEGF antibody (0.2 µg/µl) every 2.5-3 weeks for a total of 5 injections. Mice injected with an IgG control (0.2 µg/µl) and un-injected mice were used as control. Non-diabeticsiblingsofIns2(Akita)micereceivedthesametreatment.Fourweeksafter the last injection, eyes were enucleated and the different vascular plexuses wereanalysedinretinalflatmounts.CollagenIVandisolectinB4stainingwasusedto determine the percentage of the area covered by blood vessels in each vascular plexus. Central and peripheral areas of the retina were analysed separately. Results:Inthesuperficialvascularlayer,therewasaslightreductioninthevesselareaindiabeticmicewhencomparedtonon-diabeticmice.Anti-VEGFtreatmentdid not affect the vessel area in diabetic or non-diabetic mice in this layer. In the inneranddeepvascularlayers,therewerenosignificantdifferencesinthevesselarea when comparing diabetic and non-diabetic mice and anti-VEGF treatment had no effect. However, in a previous study we have shown increased albumin leakage indiabeticmicefollowingfiveanti-VEGFintravitrealinjections,suggestingthatthefunction of these vessels might be compromised while the structure remains intact. Conclusion:Ourresultssuggestthatfiveconsecutiveintravitrealanti-VEGFinjectionsdo not cause any structural changes, as assessed by vessel area, to the retinal vasculatureinIns2(Akita)diabeticandnon-diabeticmice.

11. The Synthetic Progestin Norgestrel Acts as an Antioxidant in a Model of Retinal Degeneration by Modulating Nrf2 Signalling

Ashleigh M. Byrne,AnaM.Ruiz-Lopez,SarahL.Roche,JenniferN.Moloney,AliceC. Wyse Jackson, Thomas G. Cotter

UniversityCollegeCork,Ireland

Retinitis pigmentosa (RP) is a group of highly heterogeneous inherited retinal diseases, all of which comprise progressive death of rod and cone photoreceptors via several different signaling cascades including apoptosis, autophagy and

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necrosis.Currently,diseasecausing-mutationshavebeenidentifiedin60rodphotoreceptor-genes. Despite this heterogeneity, all cases of RP are characterized by death of rod photoreceptors followed by secondary gradual death of cone cells, whicheventuallyleadstototalblindness.Althoughnotfullyunderstood,oxidativestressisakeyfactorinthemechanismofsecondaryconecelldeath.Astherodsdie, oxygen metabolism is perturbed, leading to increased retinal oxygen levels, and thus reactive oxygen species (ROS). Moreover, it has been demonstrated that ROS produced due to light exposure can potentiate the degeneration of the retina in inherited and age-related retinopathies, including RP. We have previously demonstrated the neuroprotective effects of Norgestrel, a synthetic analogue of progesterone, in both the inherited RP model, and the light-damage model of retinal degeneration. We now show that Norgestrel can also act as a powerful antioxidant in the degenerating retina. Norgestrel rescued photoreceptors from light-induced oxidative stress and subsequent cell death. Norgestrel also prevented light-induced photoreceptor morphological changes that were associated with ROS production, and that are characteristic of RP. Further investigation showed that Norgestrel acts via post-translational modulation of the major antioxidant transcription factor Nrf2; bringing about its phosphorylation, subsequent nuclear translocation, and increased levels of its effector protein superoxide dismutase 2 (SOD2). In summary, our results demonstratesignificantprotectionofphotoreceptorcellsfromoxidativestress,andunderscore the potential of Norgestrel as a therapeutic option for RP and other degenerative retinopathies, which will not discriminate between genetic etiologies.

12. Developing Novel Anti-Angiogenic Compounds for Ocular Neovascular Disorders: a Structure-Activity Relationship for a Family of Cysteinyl Leukotriene Receptor Antagonists

Alison L Reynolds,PilarVentosa,JohanGranander,ClaireKilty,ElizabethYoung, Clare T Butler, Orla Galvin, Stephanie Merrigan, Temitope Sasore, Yolanda Fernandez,DeclanGilheany,BreandánN.Kennedy.


Introduction:Previously,weidentifiedquininib(2-[(E)-2-(quinolin-2-yl)vinyl]phenol), a cysteinyl leukotriene receptor antagonist with anti-angiogenic activity in mouse models of ocular neovascularisation. Here, we perform a structure-activity relationship study to comprehensively characterise the features which confer anti-angiogenic activity. Methods:Seventyquinolineanaloguesweretested.Anti-angiogenic activity of each compound was ranked based on ability to inhibit hyaloid blood vessel formation in an in vivo ocular developmental angiogenesis assay in transgenicfli1:EGFPzebrafish.Compoundswereconsideredactivewhere≥40%inhibition was observed, compared to control. Selected highly ranked compounds

were tested in mammalian endothelial cell lines (HMEC-1) for cytoxicity (MTT assay) andanti-angiogenicactivity(invitrotubuleformationassay).Efficacywasdeterminedusingamouseexvivoaorticringassay.cysLTreceptorantagonismwasquantifiedusing reporter cell bioassays. Results:Eighteenanaloguesshowed≥40%inhibitioninformationofhyaloidvessels.Thesecompoundshadmodificationstothe“triene”bonding and benzene ring. Six compounds were selected for further testing, none ofwhichshowedasignificantreductionincellviabilityat10µMover24hours.Threecompoundsshowed≥40%reductionintubulelengthinvitroandoneshowed≥40%reductioninsproutformationinthemouseaorticringassay.Cellbioassaysshowed all six compounds inhibited LTD4 induced activation of cysLT1 (IC50: 1.7 –6.7µM)andat30µM,fourofthecompoundsshowed≥50%inhibitionofLTC4induced activation of cysLT2. Conclusions: Of the seventy compounds tested, 18 showed robust anti-angiogenic activity in vivo. Three selected quinoline compounds acting as cysteinyl leukotriene antagonists, demonstrated anti-angiogenic activity inhumancelllinesandmouseaorticring.Thisstudyisthefirststepinidentifyinga quinoline pharmacophore for antagonising the cysLT pathway as a therapeutic target for inhibiting the aberrant pathological angiogenesis occurring in ocular neovascularisation.

13. Evaluation of Potential Mitochondrial Therapies using Novel Complex I Assays

DM Maloney, N Chadderton, S Millington-Ward and GJ Farrar Institute of Genetics, Trinity College Dublin

Manydisordersinvolvingtissues,whichhavesignificantenergyrequirements,involvemitochondrial dysfunction often due to mutations affecting the mitochondrial genome. Some such mutations can involve genes coding for subunits of complex I of the electrontransportchainleadingtoacomplexIdeficiencyindisorderssuchasLeberHereditary Optic Neuropathy (LHON) amongst others. Mitochondrial dysfunction leads to a lack of energy production and ultimately the death of the cell. In disorders such as LHON, retinal ganglion cells (RGCs) are affected, leading to retinal dysfunction. These observations have prompted interest in exploring innovative therapeuticstomodulatemitochondrialdisordersinvolvingcomplexIdeficiency.TheFarrarlaboratoryhasexploredcandidategenetherapiesforcomplexIdeficiencyusingNdi1,ayeastgenewhichisacomplexIhomologue.Inordertotesttheefficacyof candidate therapies, we have developed robust, empirical assays of mitochondrial function. Some such assays measure oxygen consumption while others evaluate levelsofNADHoxidationinasample,bothbeforeandaftercomplexIinhibition(inhibitor 1) thereby providing a measure of complex I activity. To optimally distinguish betweentheactivityofcomplexIandacandidatetherapeutic,modifiedassays

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arebeingdevelopedinvolvingadditionofasecondinhibitorwhichenablesspecificmeasurement of the therapeutic, such as Ndi1. Such in vitro assays will facilitate large-scale screening of candidate therapeutics and enable only those that show strongevidenceofefficacyinvitroarethentestedinvivo.Incombinationwithotherquantitative assays such as Reactive Oxygen Species (ROS) generation, this allows detailed evaluation of the health of mitochondria within a sample.

14. A Sustained Release Formulation of Quininib-Hyaluronan Microneedles Inhibits Cysteinyl Leukotriene Induced Retinal Vascular Permeability

Orla Galvin,AkshaySrivastava,OliverCarroll,RajivKulkarni,SteveDykes,SteveVickers,KeithDickinson,AlisonReynolds,ClaireKilty,GarethRedmond,RobJones,SharonCheetham,AbhayPandit,&,BreandánN.Kennedy.

RenasciLtd,Nottingham,UnitedKingdom

Purpose: Pathologic neovascularisation and ocular permeability are hallmarks of proliferative diabetic retinopathy (DR). Current pharmacologic interventions targeting VEGF are effective in only 30-60% of patients and require multiple intraocular injections associated with iatrogenic infection. Our goal is to develop novel small molecule VEGF-independent drugs amenable to sustained ocular release, and which reduce retinal angiogenesis and vascular permeability. Here, quininib was formulated intohyaluronan(HA)microneedleswhosesafetyandefficacywasevaluatedinvivoin an acute cysteinyl leukotriene (CysLT) retinal vascular permeability (RVP) model. CysLTs are synthesised from arachidonic acid via 5-HETE which is increased in the vitreous of diabetic patients. Methods:Quininib-HAmicroneedleswereformulatedviadesolvationfromquininib-HAsolutionandsubsequentcross-linkingwith4-arm-PEG-amine prior to freeze-drying. Scanning electron microscopy revealed hollow needle-shapedparticleultrastructure,withazetapotentialof−35.5mVdeterminedbyelectrophoreticlightscattering.TheincorporationefficiencyandpharmacokineticprofileofquininibreleasedinvitrofromthemicroneedleswasquantifiedbyHPLC.RVPwasdeterminedbyEvan’sBlueleakagefromthevasculatureofBrownNorwayrats. Results: Quininib incorporation into these microneedles was 90%. In vitro, 20% quininib was released over 4 months; or in the presence of increasing concentrations ofhyaluronidase,60%incorporatedquininibwasreleasedover4months.Zebrafishhyaloid vasculature assays demonstrated quininib released from these microneedles significantly(p<0.0001)inhibitedoculardevelopmentalangiogenesiscomparedtocontrol. Sustained amelioration of RVP was demonstrated using a bespoke cysteinyl leukotrieneinducedrodentmodel.CysLTssignificantlyinduceRVPinBrownNorway rats compared to vehicle, CysLTs 30 µM vs vehicle p=0.031, CysLTs 90

µM vs vehicle p=0.005, which was greater than that observed with VEGF 50 ng vs vehiclep=0.212.Quininib–HAmicroparticlessignificantlyinhibitedRVPonemonthafter administration compared to neat quininib control (p=0.0071). Conclusion: We established a CysLT induced model of RVP in Brown Norway rats; and developed Quininib-HAmicroneedleswhichallowsustainedreleaseofquininib,aresafeinvivoand effectively inhibit angiogenesis and RVP in vivo.

15. A Novel Concept for Sub-RPE Deposit Formation Imre Lengyel,MatthewPilgrim,EszterEmri,LajosCsincsik,RichardB.Thompson,

Christine A. Curcio Queen’sUniversityBelfast,UnitedKingdom It is still not known how sub-RPE deposits in the human eye start to form. Here we

presentevidencethathydroxyapatite(HAP),thehighlyinsolublemineralcomponentof bone, is present in sub-RPE deposits and form the seeding point for the deposition of proteins and lipids both in the human eye as well as in a cellular model. For our experiments we used donor eyes that were obtained from the Eye Bank at MoorfieldsEyeHospitalorprimaryporcineandhumanretinalpigmentepithelialcells.ThesewereanalyzedbymicroprobesynchrotronX-rayfluorescence(SXRF)andx-raydiffraction(XRD)stainedwithfluorescentdyesselectiveforhydroxyapatiteandimagedbyconfocalfluorescencemicroscopy.Forimmunolabeling,tissuesandcellswereparaffin-embeddedandsectionedorflatmounted.TheXRDdataunambiguouslyidentifiedhydroxyapatiteinsub-RPEdepositsinhumaneyes.TheuseoftwoinsitufluorescentstainsforHAPshowedspherularstaininginallformsofsub-RPEdepositsandindicatedthatHAPspherulesarepresentontheBMeveninareaswheretherearenovisibledrusen.TheaveragesizeoftheHAPspherulesis~3um.Usuallynumerousspherulesaredistributedthroughoutindividualsub-RPEdepositssomethatarecoatedwithpreviouslyidentifieddrusenproteins.Whilenotidentical,similarHAPdepositionwasfoundinthecellularmodelsystemthatwascombinedwithproteinandlipidsignatures,suggestingthatHAPdepositionislikelyto be initiated by the RPE. The unexpected presence of hydroxyapatite and the closeassociationofproteinandlipidswiththeHAPspherulesinvivoaswellasinvitro points to a previously unrecognized early molecular event at the RPE/choroid interface. Understanding these molecular interactions that required for this deposit formation and growth may provide a means to forestall or reverse deposit formation well before irreversible sight loss occurs.

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16. Verification of ERG Isolation of Human Rod Photoreceptor Activity Using Silent Substitution

John Maguire1,4,DeclanMcKeefry1, Neil Parry4,JanKremers3, Ian Murray2 1BradfordSchoolofOptometry&VisionScience,UniversityofBradford,UnitedKingdom;2FacultyofLifeSciences,UniversityofManchester,UK;3Dept. Ophthalmology,UniversityHospitalErlangen,Germany;4Vision Science Centre, ManchesterRoyalEyeHospital,UK.

Purpose: To develop an ERG–based assay of human rod photoreceptor function using silent substitution stimuli without the need for dark adaptation. In order to verify the level of isolation of rod function using this method we compared ERGs elicited from normals and participants with a homozygous mutation in CNGB3 causing rod monochromacy. We also used L-, M- and S-cone isolating stimuli to record ERGs in these groups. Importantly, the examination of responses from L-, M- and S-cone photoreceptors may help provide a more complete electrophysiological characterisation of retinal function in clinical cases. Methods: Rod isolating stimuli were generated using a 4 primary LED ColorDome ganzfeld stimulator. ERGs were recorded from 5 normals and 3 participants with a homozygous mutation in CNGB3 causing rod monochromacy. In experiments 1 & 2 we examined the temporal frequency and retinal illuminance response characteristics using sinusoidal stimuli. ERGs were subjected to Fourier analysis from which the amplitude of the fundamental component was measured. In experiment 3 we examined the morphology of transient ERG waveforms elicited by rod and cone isolating stimuli withsquare-wavetemporalprofilesResults: 1) The temporal response functions forrodisolatingflickerstimuliarelow-passforbothgroupsandexhibitnoresponsebeyond 30Hz. 2) The retinal illuminance functions are similar in both groups, exhibiting maxima between 10-100 ph Td. 3) There is a high degree of similarity between the rod isolated ERGs from the normal group and those from the rod monochromats. Conclusions: Our results demonstrate that silent substitution provides an effective method for the isolation of human rod photoreceptor activity without the need for time consuming periods of dark adaption. Furthermore, the use of both rod and cone photoreceptor isolating stimuli provides the potential for a more extensive overview of function in the pathological retina.

17. Phase I Device Trial of a Next Generation “Injectable” Telescopic Lens Implant for End Stage Age Related Macular Degeneration

Kirk Stephenson,DavidKeegan MaterPrivateHospital,Dublin,Ireland

Purpose:Todescribethesafetyandefficacyofaninjectableintraoculartelescope(WA-NG:x3magnification)foruseinendstageagerelatedmaculardegeneration.Methods: Eligible patients were put through a stringent screening process. Inclusion criteria included phakic status, bilateral stable end stage macular degeneration withBCVA6/24–6/240,refractionbetween-6Dand+4D,anteriorchamberdepthof≥2.5mm,nohistoryofpreviousintraocularsurgeryandnoperipheralvisualpathology. Patients were seen up to 1 year post op with measurement of endothelial cellcount(ECC),anteriorchamberdepth(ACD)andvisualacuitymeasurement,in addition to full ophthalmic examination. Results: 3 patients were recruited and implantedwiththeinjectabletelescope(videoavailable).Allthepatientsweremale,withmeanage80.3y(range73-84y).Afteroneyear,meanECClosswas9.2%withameanresidualECCof2442cells.ACDpreopwasmean2.68mmandthisdeepenedto 3.68mm at one year. Visual outcomes were in keeping with the original IMT model, withagainin≥2linesofvisioninall3cases(range+2.6to+4.4LogMARlines).There were no serious adverse events. Discussion:Inthisphaseone(“firstintheworld”)study,theWA-NGintraoculartelescopeissafe,toleratedandachievesvisualoutcomes comparable with the original telescopic lens implant. The surgery requires a smaller corneal incision, due to the injectable format of the haptics, thus inducing less astigmatism and leading to a more rapid healing process. The haptic design allowsgreaterposteriordisplacementandthisisreflectedinECCandclearancemeasurements up to one year. Further data is required to evaluate the long term safetyandefficacyofthisdevice;howeveritisaviableassistivetechnologyforendstageAMD.

18. Standardisation of Irish Inherited Retinal Dystrophy Service Kirk Stephenson,PaulKenna,DavidKeegan MaterMisericordiaeUniversityHospitalDublin

Purpose: To investigate the current standard of practice for patients with inherited retinal dystrophy (IRD) in the Republic of Ireland versus international best practice. Method: Representatives from all public eye hospitals and a selection of private ophthalmologists within the Republic of Ireland were invited to take part in a survey. Topics covered were volumes of new/return IRD patients, availability of relevant investigations/services, and trends in management. Results: Nine centres replied tothesurvey.AllwereawareoftheTarget5000all-IrelandIRDregistry.Monthlynumbers of new/review IRD patients seen were <5 in regional centres and 6-10 in specialistcentres.Theaveragetimespentwithnewpatientswas>30minutesinallcentres. Necessary clinical tests were available in 88% of centres; 66% requested

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genetictesting.Allpatientswereseenbyophthalmologistsorgeneticists;onlyonecentre had access to a genetic counsellor (this was not full time ophthalmic genetics). 50% of centres had a low vision service available and patients were referred to NCBI low vision services from 33% of centres. Discussion: The gold standard of IRD care involves detailed phenotyping and genetic testing. Management of patients with IRD is multidisciplinary, including disease-modifying therapies (e.g. gene therapy, stemcelltherapy,artificialretinas)andmethodsformaximizingpatientfunctionandquality of life (e.g. low vision aids, social supports, genetic counselling). There is a cross-border all-Ireland IRD registry under development (Target 5000), with access to clinical and genetic testing leading to access to low vision supports and counselling (i.e. Fighting Blindness & NCBI). Seeing a higher volume of IRD patients and with the infrastructure in place to accommodate them, the specialist centres involved in Target 5000 approach international best practice. With institutional and Fighting Blindness support this is the optimal route through which to manage patients with IRD in Ireland.

19. MicroRNA Regulatory Circuits in a Mouse Model of Retinitis Pigmentosa

Arpad Palfi,KarstenHokamp,StefanieM.Hauck,SebastianVencken,SophiaMillington-Ward,NaomiChadderton,MathewCarrigan,ElodKortvely,CatherineM.Greene,PaulF.KennaandG.JaneFarrar

Trinity College Dublin, Ireland

In many human conditions, including disorders of the retina, dysfunction has been linkedtoperturbedmiRNAregulation.Forexample,wefoundanalteredexpressionof miR-1/133 and miR-183/96/182 clusters, as well as miR-142 in the RHO-P347S mouse retina, a murine model for retinitis pigmentosa. High-throughput LC-MS/MS analysis of the RHO-P347S mouse retina was used in the current study to determine theproteinexpressionofinsilicopredictedtargetsfortheabovemiRNAs;133potentialtargetswereidentified.Pathwayover-representationanalysisoftheproteinexpression signature in the RHO-P347S retina suggests G-protein signaling/visual transduction, and synaptic transmission for miR-1, and trans-membrane transport, cell-adhesion, signal transduction and apoptosis for miR-183/96/182 as regulated functions.AdditionallymiR-1,miR-96/182andmiR-96targetingofCtbp2,Rac1andSlc6a9 respectively was validated in vitro. Interaction of the miR-183/96/182 cluster andRac1mRNAintheretinawasconfirmedusinginvivoRac1-miR-CATCH.WepredictedinsilicoandvalidatedinvivothatadditionalmiRNAs,suchasmiR-103-3pandmiR-9-5ptargetRac1mRNA.UsingRac1-miR-CATCHwealsoenrichedformiRNAs,suchasmiR-125a/b-5pandmiR-378a-3p,whichwerenotpredictedinsilicototargetRac1.Insummary,ourdatasuggestsignificantutilizationofmiRNAcircuits

intheretina.WithregardtoRac1,morethan30miRNAsappeartotargetthismRNAin retina. Perturbed Rac1 expression has been implicated in retinal degenerations inanumberofstudies,mostrecentlybySongetal.(IOVS,May2016).RefiningourunderstandingoftheregulatorymechanismsinfluencingRac1expressionmayhighlight new therapeutic targets.

20. A Novel Phenotype Due to SCL24A1 Mutation Conor P. G.Malone,MatthewCarrigan,KarenCollins,HilaryDempsey,

G.JaneFarrar,PaulKenna ResearchFoundation,RoyalVictoriaEyeandEarHospital,Dublin,Ireland

Over 600 patients attending the Research Foundation have been recruited prospectively and clinically characterized during the course of a Next Generation Sequencing (NGS) initiative, (Target 5000). Patients underwent assessment of best-corrected visual acuity, Goldmann perimetry, Lanthony D15 colour vision testing, slit-lampbiomicroscopy,ISCEVstandardfull-fieldelectroretinography(ERG),colourandautofluorescencefundusphotographyandspectral-domainopticalcoherencetomography.Followinginformedconsent,DNAsamplesdrawnfromthesubjectsunderwent exon sequencing of 218 retinopathy-associated genes using target-capture oligo panels. Two siblings, the proband (52 years old) and her sister (47 years old) both reported lifelong night blindness. There was no known parental consanguinity. Both retained good central vision until their 5th decade. Goldmann perimetry showed marked concentric constriction in both patients to the IV4e, I4 and O4e targets. ERG revealed non-recordable rod-isolated responses in both patients.Cone-isolatedresponseswerenormalintimingbutverysignificantlyreduced in amplitudes. Fundoscopic examination demonstrated bone-spicule intra-retinal pigment deposits in both patients. The clinical picture strongly indicated a diagnosis of retinitis pigmentosa (RP), with likely autosomal recessive inheritance. NGS revealed that both patients were homozygous for a 1bp frameshift deletion in codon893ofSLC24A1.MutationsinSLC24A1,therodNa-Ca+Kexchanger(NCKX)gene, have been reported as causative of congenital stationary night blindness (CSNB)(RiazuddinetalAmJHumGenet.2010Oct8;87(4):523-31;NeuilléetalClin Genet. 2016 Jun;89(6):690-9). We are unaware of any reports of mutations in SCL24A1ascausativeofRetinitisPigmentosa.Werecentlyreported(CarriganetalBrJOphthalmoldoi:10.1136/bjophthalmol-2015-306939)thatamutationinGNAT1,a gene which was previously thought only to be associated with CSNB, also causes a late-onsetformofRP.OurdemonstrationofanovelmutationinSLC24A1,associatedwith classical RP, extends the range of retinal phenotypes which disruption of this gene may cause.

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21. A Variable Phenotype Associated with BBS1 Conor P. G.Malone,MatthewCarrigan,KarenCollins,HilaryDempsey,

AdrianDockery,GJaneFarrar,PaulKenna RoyalVictoriaEyeandEarHospitalResearchFoundation,Dublin,Ireland

Over 600 inherited retinal degeneration (IRD) patients have been recruited prospectively through the Research Foundation, Royal Victoria Eye and Ear Hospital, Dublin to a Next Generation Sequencing (NGS) initiative, Target 5000. We report phenotypic and genotypic results of 3 affected siblings. Each underwent assessment ofbest-correctedvisualacuity(BCVA),Goldmannperimetry,LanthonyD15colourvisiontesting,slit-lampbiomicroscopy,ISCEVstandardfull-fieldelectroretinography(ERG),colourandautofluorescencefundusphotographyandspectral-domainopticalcoherencetomography.Followinginformedconsent,DNAsamplesfromeachsubject underwent exon sequencing of 218 retinopathy-associated genes using target-capture oligo panels. The proband reported night blindness since 12 years of age,hadmarkedconcentricfield-losssymptomsatage25andunderwentbilateralcataract surgery during her 4th decade. Fundoscopy revealed extensive bone-spicule intra-retinaldeposits.AdiagnosisofRetinitisPigmentosa(RP)wasmadeat25yearsofage.HerBCVAatage45wasHandMovementsineacheye.Hersisterpresentedatage18withreduceddayvision.Moderateconcentricfieldlosswasnoted.Shereported good night vision until age 32. Fundoscopy at initial presentation showed macular pigmentary changes. Peripheral pigmentary deposits have never been observed.HerBCVAat36yearsofagewas6/15ineacheye.Abrotherreportednightblindness and compromised day vision at age 25 and underwent bilateral cataract surgeryinthe5thdecade.HisBCVAatage48was6/24ineacheye.Scatteredperipheral pigmentary deposits and macular pigmentary changes, supportive of a clinical diagnosis of RP, were observed. One other sibling, and both parents (who are not related), were phenotypically normal. NGS revealed that all 3 affected siblings werehomozygousforaT>Gtransversionatnucleotide1169inBBS1resultinginaMethioninetoArgininesubstitutionatcodon309.Met390Arghasbeenreportedascausative of approximately 80% of Bardet-Biedl Syndrome cases due to mutations inBBS1(MykytynK.etal,NatGenet.2002Aug;31(4):435-8)FollowingtheclinicaldiagnosisofRP,butpriortotheelucidationoftheBBS1Met360Argmutation,the proband was diagnosed with renal dysfunction which responded to dietary restrictions. Her sibling, with predominantly macular changes, was diagnosed with non-insulin dependent Diabetes Mellitus, controlled on diet. None of the individuals wassignificantlyobeseandnonehadanyevidenceofcompromisedmentation.Direct questioning revealed that 2 of the 3 siblings had supernumerary toes removed in early childhood. Pleotropism in Bardet Biedl Syndrome is well documented. There is marked genetic heterogeneity, with approximately 14 genes to date implicated in the condition. BBS1 mutations are thought to account for approximately 25% of cases of the Syndrome. This study indicates that even within the same family the condition maypresentdifferentlyinaffectedsiblings.Theidentificationofawell-knownBBS1mutation in patients in whom the clinical diagnosis did not initially point to Bardet BiedlSyndromeemphasisestheutilityofgenotypinginhelpingtorefinetheclinicaldiagnosis.

22. Investigating a Role for Aberrant Epigenetic Regulation in Genetic Eye Disease: Preliminary Approaches in a Complex Phenotype, Dry AMD

Louise F Porter,EikodeJong,RenskeKuiper,MarcelCoolen,PaulNBishop,LakisLiloglu,SimonClark,AnnekedenHollander

UniversityofLiverpool,UnitedKingdom

Introduction:Age-relatedmaculardegeneration(AMD)isthecommonestcauseofvisionlossintheUK.Itiscausedbybothgeneticandenvironmentalinfluences,although the link between the two factors is sometimes tenuous. Genetic approaches haveidentifiedmajorcommonvariantsconferringAMDriskonchromosomes1q31 (complement factor H (CFH) (Y402H)) and 10q26 (age-related maculopathy susceptibility2(ARMS2)andadjacenthigh-temperaturerequirementfactor(HTRA1)genes).Overall19lociconveyingsusceptibilitytoAMDareknown,estimatedtoaccount for 65% of its heritability. It remains unclear what effect these SNPs have on gene expression levels overall. Epigenetics has been implicated in mediating, in part, the disease risk associated with some environmental factors, therefore we investigateapossibleepigeneticcontributiontoAMD.DNAmethylation,particularlyat CpG dinucleotides, is an important mechanism of gene silencing. Differential CpG dinucleotideDNAmethylationlevelsinthevicinityofAMDrisklocimayresultinaltered gene expression levels of target genes. We are performing an epigenome-wideassociationstudy(EWAS)aimingtosystematicallyuncovertheassociationbetweenepigeneticvariants(inthiscaseDNAmethylation)anddisease,potentiallyenablingtheidentificationofdisease-associateddifferentiallymethylatedregionsandlocifordryAMD.Methods:2-stageGenome-widemethylationanalysisofDNAsamplesisolatedfromdonoreyes(24dryAMD,24controls-discoverycohort;24dry,AMD,24controls-replicationcohort)usingIllumina450Karrayprofilingof485000methylationsites.SitesthataredifferentiallymethylatedinAMDversuscontrolRPEcell-derivedDNAinboththediscoveryandreplicationcohortswillbeanalysedusingpyrosequencing(methylation-specificPCR).Thecollectionofgoodqualityreplicated data will also enable incorporation of this study into future meta-analyses.

23. Locus Resequencing of ABCA4 in STGD1 Patients Uncovers Novel Deep-Intronic Splicing and Putative Cis-Regulatory Mutations

Miriam Bauwens, Riccardo Sangermano, Nicole Weisschuh, Frans Cremers, Timothy Cherry and Elfride De Baere

GhentUniversity,Belgium

Purpose:AutosomalrecessiveStargardtdisease(STGD1)ishallmarkedbyalargeproportionofpatientswithasingleheterozygouscausativevariantinABCA4.AfterBraunetal.(2013)reporteddeep-intronicABCA4variants,weresequencedtheentireABCA4geneanditscis-regulatoryregionin116BelgianandGermanSTGD1patientswithoneornoABCA4varianttouncovernovelnon-codingABCA4variants.Methods:AfterenrichmentwithacustomizedHaloPlexpanelthatincludestheentireABCA4geneanditscis-regulatorydomainaswellasPRPH2,all116patientsunderwenttargetedlocusresequencingusingNGS.Variantfilteringwasbased

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on criteria such as minor allele frequency, frequency in the in-house sequenced population, splicing predictions and regulatory potential as determined by their presenceinretina-specificcis-regulatoryregions(T.Cherry,unpublisheddata)andregulomeDB datasets. Mini-gene assays were conducted to functionally validate putative splicing variants, while luciferase assays were performed to assess potential cis-regulatory effects. Results:TargetedresequencingofABCA4andPRPH2revealedaputativepathogenicorpathogenicvariantin~50%ofcases.SomeoftheABCA4variantswerecodingvariantsofunknownsignificance;intronicvariantsclose to intron-exon boundaries, the pathogenicity of which was demonstrated by mini-gene assays, or already published deep-intronic variants. Interestingly, a novel deep-intronicvariantinintron30wasidentifiedintwounrelatedpatients,ofwhichmini-gene assays demonstrated the inclusion of a cryptic out-of-frame exon. Finally, weidentifiedputativecis-regulatoryvariantsinthe5’UTRofABCA4.Usingluciferaseassayswedemonstratedasignificantlyreduced(p<0.01)activityofoneofthesevariants. Conclusions: Targeted resequencing in 116 STGD1 patients revealed novel deep-intronic splicing and cis-regulatory mutations, for some of which the functional effect could be demonstrated. Our study illustrates the need for targeted locus resequencing in STGD1 patients.

24. Target 5000: Genetic Characterisation of a Cohort of Inherited Retinal Degeneration (IRD) Patients

Adrian Dockery,MatthewCarrigan,ConorMalone,DavidKeegan,KirkStevenson,JulieSilvestri,AndrewGreen,JohnMcCourt,PeteHumphries,PaulF.Kenna, G. Jane Farrar

Trinity College Dublin, Ireland

The Target 5000 project is focused on genetic characterisation using target capture next generation sequencing (NGS) of a cohort of Irish patients with an inherited retinal condition, estimated to be about 5,000 individuals. Many clinical trials are being undertaken for inherited forms of ocular disease, however, many such trials require patients to have their causative mutation known and validated in order to be eligible for consideration as participants in trials. The aim of the study is to genetically identify patients with inherited retinal degenerations (IRDs) in Ireland, to characterise disease causing mutations and in principle potentially enable clinical trials to be more accessible to some patients where appropriate. The study also seeks to detect formerly undiscovered pathological mutations in the target capture panel of known IRD genes under evaluation. To date as part of Target 5000 approximately 10% of the Irish IRD population has been sequenced for 220 IRD genes providing an initial informative snapshot of retinal degeneration in Ireland. Target 5000 offers not only a chance to discover new relevant and pathogenic mutations, but is vital in giving patients access to information regarding the genetic pathogenesis of their disease. Over 50 novel mutations have been discovered, as well as some previously ambiguous phenotypes resolved. Greater knowledge regarding genotype and phenotype of IRDs from this study, and similar studies globally, should provide a greater understanding of the natural histories of these disorders and should aid in development of more robust future diagnoses and prognoses.

25. Analysis of Photoreceptor Pointing Using Quadrant Pupil Detection Salihah Qaysi, Denise Valente, Brian Vohnsen UniversityCollegeDublin,Ireland

Photoreceptors are endowed with directional properties that are expressed by their angular sensitivity to incident light as well as directionality in retinal images commonly referredtoastheStiles-CrawfordeffectofthefirstkindandtheopticalStiles-Crawford effect, respectively. Individual photoreceptor tilt can be analysed by moving the incident light across the pupil when capturing retinal images. The aim of this study is to examine the viability of a quadrant pupil detection scheme in which light enters near the SCE peak and backscattered light is captured through four equal-sized sectors in the pupil from which individual photoreceptor tilt can be derived. The method employs a pyramidal prism to capture simultaneously 4 high-resolution retinal images.AnumericalanalysisusingMatlabisperformedtoquantifytheangulartuningin scattered light from each cone in the captured retinal images. The experimental results for parafoveal retinal imaging in healthy subjects are compared with modeled cone mosaic images. Retinal photoreceptors are modeled by single-layered scattering by mitochondria in the ellipsoid. Retinal images are calculated through four equal-sized sectors in the pupil lane in a 4f system. The Modeled photoreceptors are placed in the retinal plane with different angular tilts. The initial result of our sectored quadrant pupil imaging system allows not only determination of total intensity images, but also direct determination of photoreceptor inclinations in the backscattered light intensity. It is found that the method is highly suited to determine photoreceptor inclinations without requiring displacement of the incident light in the pupil plane, wherebythedeterminationofphotoreceptortiltbecomessimplified.Theexperimentalresultscomparedwellwiththetheoreticalexpectationstherebyconfirmingthepotential of the technique. It offers the possibility to analysis cone photoreceptor tilt in 2-D and is expected to probe valuable when analysing retinal disease.

26. Assessing Cone Photoreceptor Structure in Patients with Exon 3 Interchange Haplotypes in the OPN1LW/OPN1MW Gene Array

Emily J. Patterson,ConorMalone,KarenCollins,AndrewGreen,MatthewCarrigan,JaneFarrar,MaureenNeitz,PaulKenna,JosephCarroll

MedicalCollegeofWisconsin,USA

Purpose: Mutations in the OPN1LW and/or OPN1MW genes lead to a wide range of phenotypes–fromred-greencolourvisiondeficiencytoprogressiveconedystrophy.Advancesinhigh-resolutionretinalimaginghaveenableddetailedexaminationof cone structure in the living human retina, which is of critical importance in assessingtherapeuticoptionsinthesepatients.HereweusedAOSLOandOCTtocharacterize cone structure in two brothers with mutations in both their OPN1LW and OPN1MW genes. Methods: Two males (aged 14 and 19 years) with suspected blue cone monochromacy (BCM) were recruited from the Royal Victoria Eye & Ear Hospital. The OPN1LW/OPN1MW gene array was characterized and the thickness and integrity of the photoreceptor layers were evaluated using OCT. The cone mosaicwasimagedusingAOSLO(bothconfocalandnon-confocalsplit-detection).

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Cone density was measured at foveal and parafoveal regions of interest using semiautomated cone-counting software. Retinal imaging was performed at the AdvancedOcularImagingProgramattheMedicalCollegeofWisconsin.Results: BothsubjectshadtheLIAVAexon3haplotypeencodedbytheirOPN1LWgene,withMIAVAencodedbytheirOPN1MWgene.OCTimagingrevealedreducedretinalthickness, while parafoveal cone density was around 24% of normal for the older brother and 34% for the younger brother. Conclusions:HerewepresentthefirstAOSLOimagesofpatientsharboringtheMIAVAhaplotype.Weobservedevidenceof remnant cone inner segment structure in both males, which is consistent with previousobservationsinpatientswiththeLIAVAhaplotype.Whileconfocalimagesrevealed reduced numbers of waveguiding cones, the overall appearance of the mosaic was suggestive of a contiguous arrangement of inner segments at the fovea, in stark contrast to previous observations in BCM. This study highlights the need for increased access to genotyped patients for advanced retinal imaging studies.

27. Multimodal Retinal Imaging in Dementia Lajos Csincsik, Timothy Shakespeare, Sebastian Crutch, Tunde Peto, Imre Lengyel Queen`sUniversityBelfast,UnitedKingdom

Purpose: Pathological changes in the eye have been reported in a range of neurodegenerativediseases.Changesoftheretinalnervefiberlayer(RNFL)dueto ganglion cell loss, retinal vessel caliber as well as increased accumulation of extracellular drusen deposits had been associated with dementia related changes in thebrain.HerewereportonthebaselinecharacteristicsofpatientswithAlzheimer`sdisease(AD)andPosteriorCorticalAtrophy(PCA).Methods: Optical coherence tomography(OCT)imageswereacquiredof72healthycontrols(HC;MMSE>28),22AD(MMSE<20)and27PCA(MMSE<24)patientsusingOPTOSSD-OCT/SLO.The study had full local Ethical Committee approval. The images were analysed for macular volume (MV) and thickness (MT), peripapillary (pp) RNFL, RPE-Photoreceptor, ONL, OPL-INL and whole retinal (WR) thickness as well as overall peripapillaryvesselcaliber(ppVC).StatisticalanalysiswascarriedoutusingSTATAand SPSS. Images with poor quality were removed from analysis. Results: There wasnosignificantagedifferencebetweenHC,ADandPCApatients(66.2±7.4vs64.9±6.5vs65.5±7.8;p>0.1).WefoundsignificantreductioninppOPL-INLthickness(p=0.006)inpatientswithPCAcomparedtoHCandAD,andsignificantincreaseinppVCinAD(p<0.001)andPCA(p=0.032)comparedtoHC.Whilewefoundnosignificantdifferences(p>0.05)betweenPCA,ADandHCinanyotherOCTmeasuresoveralltherewasasignificantagerelateddecreaseonallofthoseparameters. Conclusions: Our results suggest that analysing changes in all retinal layers might be necessary to obtain a full picture of the pathology in the retina associated with dementia. In addition, retinal vessel caliber segmentation might hold relevant information in regards to dementia associated pathologies. While therearechallengesinimagingpatientswithADandPCAnewimagingmodalitieshave the potential to reveal previously unrecognized association between retinal manifestations of dementia.

28. Human Retinal Imaging Using Scanning Laser Ophthalmoscope Prince Sunil Thomas and Brian Vohnsen Advanced Optical Imaging Group, School of Physics,


Ascanninglaserophthalmoscopehasbeenusedtoimagetheretinainthenormalhuman eye. To resolve individual cone photoreceptors near fovea centralis, the system was aligned with minimal aberrations but without using adaptive optics. Afocusedinfraredscanninglaserbeamrasterscanstheretinaandimagesarecaptured confocally in a conjugate plane with an avalanche photodiode. Individual cone-photoreceptor directionalities has been measured using the scanning laser ophthalmoscope. The brightness of cone cells is due to the elevated refractive index of the mitochondria where the incident beam gets backscattered. Retinal scans were also able to resolve the blood vessels in the retina. The cone directionality is determined at different retinal eccentricities where the cones are estimated to have a diameter of about 5-10 µm, matching approximately to the spot size of the incident beam. Finally, measured cone directionality values are compared with the predicted directionalitiesfromthecouplingefficiencytophotoreceptorwaveguidemodesaswell as from expectations based on psychophysics. With the integration of adaptive optics into the present system, a greater resolution of cone cells is expected. This also leads to the possibility of increase in the resolution of blood vessels with a potential to identify micro aneurysms particular to diabetic retinopathy. Retinal scans obtained using the scanning laser ophthalmoscope could reveal the packing arrangement of cones in the transverse plane. Difference in the packing arrangement or structure of the cones will be an indicator of a retinal disease. The present study paves the way for a possible use of scanning laser ophthalmoscopy to target cone-specificretinaldiseases.

29. Baseline Characteristics of Patients with Neovascular Age-related Macular Degeneration from the Real-world LUMINOUS Study: Global vs Irish Results

Coppins, Jennifer5;Beatty,Stephen1;Cahill,Mark2;Henry,Emer3;Keegan,David4; 1InstituteofEyeSurgery,WhitfieldClinic,Co.Waterford,2BeaconClinicandHospital,

Sandyford,Dublin;3WaterfordRegionalHospital,Waterford;4MaterPrivateHospital,Dublin;5Novartis Ireland Limited, Dublin

Study Group: Luminous

Purpose: LUMINOUS (NCT01318941) is a 5-year multicenter, observational study to evaluate the long-term safety, effectiveness, treatment patterns and health-related quality-of-life associated with ranibizumab (RBZ) treatment in clinical practice for all licensed indications. Here, we describe the baseline characteristics of the enrolled globalandIrishnAMDpatients.Methods:Ofthe29052adultpatientsenrolledgloballypriortoMarch2014,364(1.25%)wereIrish:245(67.3%)werenAMDpatients. Both treatment naïve and prior RBZ patients were enrolled (19 vs 226). Results:ThemeanageofIrishnAMDpatientswashigherthantheglobalcohort(75.8yrsvs74.5yrs).Overall,baselineVAwashigherintheIrishcohortthantheglobal cohort (treatment naïve: 60.4 vs 49.3, prior-RBZ: 62.5 vs 56.3, respectively). In

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theIrishcohortbaselineVA(ETDRSletterscore)washigherinthepriorRBZpatientsthan in the treatment naïve group (62.5 vs 60.4). Non ocular comorbidities were lower in Irish patients with the exception of hyperchloesterolemia / lipidemia (51.0% vs 30.4%) and history of coronary artery disease (36.3% vs 13.9%).The mean CRT at baseline of Irish patients was lower than the global cohort (201.8µm vs 303.6 µm). The median time to diagnosis in years was longer in Irish patients than in the global cohort (1.503 vs 0.869). There were more females in the Irish cohort (71.8% vs 57.6%). Conclusions: LUMINOUS includes patients with more diverse demographics than typically reported for clinical trials, and is more representative of the real world patient population. Overall, baseline demographics were similar except gender and race,whencomparedtoglobalaverages.AlsopriorRBZtreatmentwasassociatedwithnumericallyhigherVAthanthetreatmentnaïvegroup.AnalysesofLUMINOUSstudy data are expected to provide substantial evidence on the long-term safety, effectiveness, and treatment patterns of ranibizumab in routine clinical practice.

30. The Use of 3D Animated Video for Learning Sean Kirwan1,CaoimheKeoghegan1,DrBreandánKennedy1, Dr Rosemarie

Carew2,TomásO’Sullivan2, Carlos Tena2, Dr James Corcoran2,DrPaulHalley2, Dr Omar Mahfoud2.

1UniversityCollegeDublin,Ireland 2ICON Firecrest

Goal: The goal of this outreach initiative is to provide a concise, accessible and engaging platform, to enhance public and patient awareness of different forms of vision loss. 3D animated videos were completed on i) age-related macular degeneration; ii) diabetes; iii) diabetic retinopathy; and iv) inherited blindness. Method: Firstly, focus-group meetings involving several key stakeholders were set-up to consider and evaluate the content to be included in the animations. Next, scripting ofthevideoswascarriedoutwhichoutlinedscene-by-scenethevideos’contentand voiceover narration. Subsequently, a storyboard document was designed, whichdefinedindetailthegraphicalinstructionforthegraphicdesigners,tocreatethe 3D animation videos. Results: The 3D animated videos average 5 minutes in length; combining a blend of visual animations and audio voiceover; which are freely accessible for researchers, clinicians and patient advocates to use in lectures, seminars, consultations, websites, social media and public engagement events. Each of the four animated videos was publicly disseminated through a variety of social medianetworks,patientadvocatewebsites,scientificresearcherforums,publicawareness blogs and secondary school visits. The 3D animated videos have over 7,600 views, reaching 14,500 people internationally through Facebook alone. From a total of 257 surveys carried out, over 98% of people agreed that this type of content isverybeneficialforincreasingawareness,andover51%saidtheywouldprefertolearn using 3D animated videos instead of written or still picture learning. Conclusion: The use of short concise 3D animated videos is a novel method of disseminating awareness for disease areas and enhances teaching and learning for all disciplines of science and non-science backgrounds. Thus, it can be concluded from the above that the use of 3D animated videos is an effective method of raising awareness and understanding of disease conditions to an extensive audience.

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