2017 ANNUAL REPORT - ORIAoria.org.au/wp-content/uploads/2012/02/ORIA-Annual-Report-2017.pdf · Her key achievements include co-ordinating the ORIA’s annual grant ... 8 ORIA Annual

THE OPHTHALMICRESEARCH REPORTINSTITUTE OF AUSTRALIA

O R I AAdvancing eye research

2017 ANNUAL REPORT

2017 ANNUAL REPORT

CONTENTSNotice of Meeting 4

Chair’s report 6

ORIA Grants 8

Progress reports 11

Directors’ Report 42

Financial Statements 46

Auditors’ Report 65


4 ORIA Annual Report 20174 ORIA Annual Report 2017

NOTICE OF MEETINGTHE ANNUAL REPORT WILL BE PRESENTED AT THE SIXTY FIFTH ANNUAL GENERAL MEETING TO BE HELD IN PERTH, WA ON SUNDAY 29 OCTOBER 2017

THE BOARDClinical Prof. Stephanie Watson, Sydney (Chairman)

Prof Mark Gillies, Sydney (Vice Chairman)

Prof Richard Mills, Adelaide (Honorary Secretary)

A/Prof Paul Healey, Sydney (Honorary Treasurer)

Dr Fred Chen, Perth

Prof J Crowston, Melbourne

Dr Clare Fraser, Sydney

Dr Jennifer Fan Gaskin, Melbourne

Prof Stuart Graham, Sydney

Dr Gerald Liew, Sydney

Prof David Mackey, Perth

Prof Paul Mitchell, Sydney

Prof Peter McCluskey, Sydney

Dr John Males, Sydney

A/Prof Andrea Vincent, New Zealand

Dr Andrew White, Sydney

5Notice of Meeting 5

HONORARY SECRETARY

RESEARCH ADVISORY COMMITTEE

For assessments of funding in 2017

Prof David Mackey, Perth (Chair)

A/Prof Alex Hewitt, Hobart (Secretary)

Prof Stuart Graham, Sydney (ex officio Chair ORIA)

Dr Kathryn Burdon, Adelaide

Dr Fred Chen, Perth

Prof Jamie Craig, Adelaide

Prof Jonathan Crowston, Melbourne

Dr Samantha Fraser-Bell, Sydney

Dr Chris Layton, Brisbane

A/Prof Ian Trounce, Melbourne

Dr Peter Van Wijngaarden, Melbourne

A/Prof Andrea Vincent, New Zealand

Dr Trevor Sherwin, New Zealand

Clinical Prof. Stephanie Watson, Sydney

Save Sight Society NZ rep – Dr Graham Wilson

Anne Dunn Snape (as Executive Officer, ORIA)

Prof Richard Mills

94-98 Chalmers Street,

Surry Hills NSW 2010

HONORARY TREASURER

A/Prof Paul Healey

94-98 Chalmers Street,

Surry Hills NSW 2010

TRUSTEES

HON SOLICITORS

EXECUTIVE OFFICER

ACCOUNTANTS

AUDITORS

McLean Delmo Bentleys

302 Burwood Road,

Hawthorn, VIC

Orr Martin & Waters

461 Whitehorse Road,

Balwyn VIC 3103

National Australia Trustees Ltd,

Melbourne VIC

King and Wood Mallesons

Advance Bank Centre

60 Marcus Clarke Street, Canberra 2600

Ms Anne Dunn Snape, BA

(Soc & Pol Phil), (MQ)

PostGradC Ethics & Legal Studs

The Ophthalmic Research Institute of Australia is the College’s research arm, and aims to: “Advance Eye Research”. The ORIA held its first AGM over 60 years ago and since that time has distributed literally millions of dollars to provide funding for medical eye research in Australia.

The ORIA’s activities are co-ordinated and managed by up to 16 members of the Board of the ORIA and Executive Officer, Anne Dunn Snape. Using the income from its investments and donor organisations, the ORIA continued to contribute to funding for research projects throughout Australia.

The ORIA received 52 applications from Australian researchers during 2016 for funding during 2017 which were subsequently assessed by the Research Advisory Committee. The ORIA’s 17 member Research Advisory Committee is composed of leading research scientists and ophthalmologists from Australia and New Zealand. The recommendations of the Committee are put forward to the Board of the ORIA who then indicate what funds are available for the forthcoming calendar year.

With available funding, the ORIA is supporting 11 projects during the 2017 funding year. The total amount provided for medical eye research by the ORIA and its supporters is just under $800,000. We are thankful to the New South Wales Branch of RANZCO and the Eye Surgeons’ Foundation for their support, as well as previous benefactors.

At the close of business April 20 2017, the ORIA received applications from 54 research groups throughout Australia for assessment for funding in 2018. These are currently being peer reviewed by experts in the field from Australia and internationally.

The ORIA assists the Save Sight Society of New Zealand by incorporating their applications for funding into the ORIA’s assessment process which is conducted from April to September.

The ORIA also continues its annual support of the Ringland Anderson Chair of Ophthalmology in Victoria.

CHAI

R’S

REPO

RT

6 ORIA Annual Report 2017

The ORIA is always mindful of auditing its research funding to assess how well its mission to advance eye research is being achieved. The results of funding support during 2016 are exciting. A financial statement from each project/institution is also secured to ensure funds have been used in the manner previously indicated in the application.

The progress/final reports printed in the annual report this year not only highlight the results but also draw on the implications for both clinical practice and science for the ophthalmology profession and its patients.

Details of all other grants awarded can be found on the ORIA website www.oria.org.au and for New Zealand at www.safesightsociety.org.nz.

In October, we say farewell to our Executive Officer, Anne Dunn Snape after 15 years with the ORIA. During the last 15 years, Anne has been as asset to ORIA. Her key achievements include co-ordinating the ORIA’s annual grant process which has seen an increase from 18 applications to 55, reviewing and preparation of the new ORIA Constitution with King Wood and Mallesons, design and preparation of the annual reports including showcasing progress reports from awardees, helping to build the much needed funds to support ophthalmic research, as well as all aspects of the management of the organisation. Anne leaves at a time when the equity of the ORIA has increased 20 per cent during the financial year and the Richard and Ina Humbley Foundation, of which she fostered, has now been established with the ORIA as its sole beneficiary. Anne is leaving ORIA in a stronger position than when she started, she will be missed by all at ORIA. I would personally like to wish her the best in her future endeavours.

Stephanie Watson

Chair, ORIA

Anne Dunn Snape

Executive Officer, ORIA

The ORIA is currently receiving progress reports for researchers funded during 2016. At this early stage, two of the researchers have received NH&MRC funding based on the initial support received by the ORIA. The two projects are

ORIA/G J WILLIAMS BEQUEST GRANT Prof J Smith ICAM-1 expression in retinal endothelial cells

ORIA/EYE SURGEONS’ FOUNDATION GRANT Dr R Natoli, A/Prof R Essex & Prof J Provis MicroRNA provide a new class of therapeutics for diseases such as Age-Related Macular Degeneration

ORIA grants have also had translational impact. Another research group has a Patent pending based on the results generated from the ORIA funding support.

Supporting new investigators is a key ORIA activity. Three new investigators now have a grant track record after receiving ORIA funding during the year. At least one has applied for additional funding based on data available from their ORIA funded research.

At least 22 publications and 7 presentations have arisen out of this year’s research funding.

7Chair’s Report


ORIA PROJECTFUNDING 2017

ORIA / ESME ANDERSON GRANTDR GUEI-SHEUNG LIU, A/PROF ALEX HEWITT,

A/PROF BANG BUI, DR ANNA KING &

DR AMY FAN LI

Comparison of different proteins

for direct gene editing in the retina

$50,000

ORIA / RICHARD AND INA HUMBLEY FOUNDATION GRANTDR FRED CHEN

Stem Cell Therapy for Age

Related Macular Degeneration

$50,000

ORIA / BRENDA MITCHELL GRANTDR SANDY HUNG &

MRS SANDRA STAFFIERI

Using patient-derived stem cells

to model retinoblastoma

$50,000

ORIA/EYE SURGEONS’ FOUNDATION GRANTDR VIVEK GUPTA & PROF STUART GRAHAM

PHLPP as a novel target to protect

retina against glaucoma damage

$49,700

ORIA/EYE SURGEONS’ FOUNDATION GRANTPROF JUSTINE SMITH

Interactions between Mononuclear Phagocytes

and Endothelial Cells in Human Retina

$49,850

ORIA / RANZCO NSW BRANCH GRANTA/PROF R MAX CONWAY, DR SVETLANA

CHEREPANOFF, DR MICHAEL GIBLIN,

PROF RICHARD EPSTEIN, A/PROF

ANTHONY JOSHUA, DR WENCHAN WONG

& MS AMPARO HERRERA-BOND

Developing technology to improve

healthoutcomes in primary eye

melanoma patients

$49,850

9Project Funding 2017 9

DONATIONS:RANZCO, NEW SOUTH WALES BRANCH

THE EYE SURGEONS’ FOUNDATION

THE ESTATE OF HARDIE-ANSELMI TRUST

THE ESTATE OF THE LATE MARY HELENE TILDEN

THE RICHARD AND INA HUMBLEY FOUNDATION

ORIA / GLADYS CLARE DICKSON GRANTDR MICHELE C MADIGAN &

PROF PETER J MCCLUSKEY

Eye melanocytes and inflammation

$46,600

ORIA NEW INVESTIGATOR/ ANSELMI BEQUEST GRANTDR SRUJANA SAHEBJADA

Genes in Keratoconus corneas

$49,105

ORIA NEW INVESTIGATOR/ RICHARD AND INA HUMBLEY FOUNDATION GRANTDR JIA JIA LEK

Reticular pseudo-drusen (RPD):

A high-risk factor for vision loss in

age-related macular degeneration (AMD)

$49,105

ORIA / RENENSSON BEQUEST GRANTDR YUYI YOU &

A/PROF ALEXANDER KLISTORNER

Trans-synaptic degeneration in the visual

pathways in glaucoma: An MRI Study

$48,450

ORIA NEW INVESTIGATORDR MICHELLE SUN, A/PROF

ANDREA O’CONNOR & DR JOHN WOOD

Bioengineered Eyelids

$47,850

TOTAL FUNDED

$539,205WWW.ORIA.ORG.AU


A Martins

Abi Tenen

Adrian Fung

Ajay Kumar

Ajoy Vincent

Alex Holcombe

Alex P Hunyor

Allison McKendrick

Andy Chien

Aparna Jayachandran

Balamarali Ambati

Bang Bui

Brian Sloan

Colin Willoughby

Darren Kelly

Donald Mutti

Dr. Damien Harkin

Fay Khong

Fred Chen

George Black

George Kong

Gerald Liew

Glyn Chidlow

Greg Dusting

Gwyneth Rees

Heather Connor

Heather Mack

Hemal Mehta

Hitesh Peshavariya

Holly Chinnery

Ilva Rupenthal

Isabella Cheung

James Bourne

James Elder

James Wong

Jason Steel

Jennifer Arnold

Jenny Berka

Jie Zhang

John Forrester

John Foster

Julie Lim

Justine Smith

Karl Brown

Katie Edwards

Katrina Schmid

Keryn Williams

Kevin Spring

Liesel Fitzgerald

Ling Zhu

Lyndell Lim

Marc Sarossy

Mark Wilcox

Matthew Simunovic

Michael Jones

Michelle Madigan

Mirella Dottori

Mo Dirani

Mona Awadalla

Monica Acosta

Nicholas Blackburn

Nick Di Girolamo

Nigel Barnett

Nigel Morlet

Owen Marshall

Paul Baird

Paul Donaldson

Peter Hadden

Petra Liskova

Ray Wong

Rebecca Pelekanos

Richard Mills

Robert Casson

Robyn Guymer

Robyn Jamieson

Rohan Essex

Rohan Merani

Russ Van Gelder

Samuel McLenachan

Sasha Klistorner

Scott Read

Shiwani Sharma

Svetlana Cherepanoff

Thomas Campbell

Tom Edwards

Verity Oliver

Vishal Jhanji

Vivek Gupta

Zhi Chao Wu

Zhichao Wu

THANK YOUTHE INSTITUTE WOULD LIKE TO THANK OUR EXTERNAL REFEREES WHO KINDLY GAVE ADVICE WHICH HELPED WITH THE ALLOCATION OF THE 2017 GRANTS. THEIR WORK IS INVALUABLE.

PROGRESSREPORTS

PROGRESS REPORT


OVERVIEW

This study aimed to use stem cells to model eye diseases. To study diseases with complex genetics (such as glaucoma or AMD), a large cell population is needed to obtain the right power of analysis, which would require a much higher number of staff. This bottleneck could be addressed through the use of robotics, which could maintain hundreds of samples, opening up new avenues into the research of complex diseases.

AIM

The overall aim of this work was to adapt our current maintenance and differentiation protocols to an automated platform to dramatically increase sample size, reduce variability and thus and allow for future high-throughput analysis of the transcriptome and metabolome in retinal cells derived from patient iPSCs. We have been successful in this adaptation and our work resulted in two publications (enclosed).

RESULTS

We manually reprogrammed 77 skin fibroblast lines from individual patients to iPSCs. All lines were successfully passaged on the platform. We then directed the differentiation of human PSCs towards two retinal lineages, retinal pigment epithelium cells and retinal ganglion cells. These results provide proof-of-concept that automation can be utilised to facilitate stem cell maintenance and retinal differentiation. Importantly, no contamination was observed during the differentiation procedure, demonstrating the robustness of the automated platform for long-term sterile cell culture.

In summary, we report the successful maintenance, passaging and differentiation of human PSCs, using an automated platform equipped with liquid handler and robotic arms, to handle cells in tissue culture plates. By increasing sample size and reducing variability, it allows for more defined parameters for future high-throughput analysis of the transcriptome and metabolome of progeny cells derived from patient iPSCs. This will be particularly important for iPSC modelling of complex genetic diseases, such as glaucomas and age-related macular degeneration, which will require large sample sizes to provide sufficient power for statistical analysis.

ORIA/ANSELMI BEQUEST GRANT

DR R WONG, A/PROF PÉBAY AND MR D CROMBIE

AUTOMATED DIFFERENTIATION OF PATIENT DERIVED STEM CELLS INTO EYE CELLS

13Progress Report

Publications (* Equal authors)

1.Crombie DE*, Daniszewski M*, Liang HH, Kulkarni T, Li F, Lidgerwood GE, Conquest A, Hernandez D, Hung SS, Gill KP, De Smit E, Kearns L, Clarke L, Sluch VM, Chamling X, Zack DJ, Wong RCB, Hewitt AW* and Pébay A*. Development of a modular automated system for maintenance and differentiation of adherent human pluripotent stem cells. SLAS Discovery. Accepted 25.01.17. [IF: 2.218; 5y: 2.003].

2.Daniszewski M, Crombie DE, Henderson R, Liang HH, Wong RCB, Hewitt AW and Pébay A*. Automated cell culture systems and their applications to human pluripotent stem cell studies. SLAS Technology. Accepted 06.05.17. [IF: 1.297; 5y: 1.563].

An application for a Patent arising out of this research was submitted by the investigators:

Automated system for maintenance and differentiation of pluripotent stem cells.

Australia and Germany application number: 2017100376;

Application date: 04.04.17

Inventors: Alice Pébay, Duncan Crombie, Alex Hewitt, Helena Liang, Raymond Wong, Maciej Daniszewski.

Ray Wong, Alice Pebay & Duncan Crombie and project team at CERA together with Mr Peter Clemenger whose Foundation also provided financial assistance

PROGRESS REPORT


OVERVIEW

We hypothesized that functional assessment of trabecular meshwork (TMW) cells derived from patients with differing ABCA1-POAG risk profiles will provide essential insight into disease mechanisms. Due to the extreme difficultly in obtaining TMW cells from living donors, we proposed to generate induced pluripotent stem cells (iPSCs) from POAG patients with specific genetic backgrounds as well as control lines. These cells would subsequently be differentiated into TMW cells to assess their functions, identify differences and dissect the ABCA1 allele-specific signalling pathways involved.

AIM

The Specific Aims of this proposal were:

Aim 1: To generate iPSCs from people who are homozygotic low or high risk at the ABCA1 locus.

Aim 2: To functionally characterise patient-specific TMW cells. We will establish the basic biology of these iPSC-derived TM cells in terms of cholesterol efflux, phagocytic ability and responsiveness of MYOC secretion to steroid treatment.

RESULTS

We have generated iPSCs from individuals with POAG and with differing ABCA1 allelic risk profiles using episomal transfer of pluripotency factors. We have also characterised their pluripotency and karyotype, and have identified clones that are suitable for further study. We have also devoted substantial time to generating quantitative assays that will enable a robust assessment of TMW cell function, and which are widely used in the field. To this end, we have developed assays to quantify MYOC secretion using ELISA, contraction of collagen gel substrates, and phagocytosis

Despite obtaining promising preliminary data using human embryonic stem cells and following publishing protocols, our attempts to generate TMW from iPSC have thus far proven to be variable and have prevented an analysis of ABCA1 function. However, in parallel experiments using mesenchymal stem cells, we have identified that combinatorial exposure to retinoic acid, TGF-β2 and BMP4, factors that each has a role in TMW cell development, caused a synergistic induction of MYOC gene expression, resulting in significant secretion of MYOC into the culture medium. This work is currently being prepared for peer review, and may ultimately contribute to refinement of protocols for TMW cell generation from iPSCs. Our future work will focus on improving approaches to TMW cell differentiation from iPSCs, prior to undertaking an interrogation of how ABCA1 risk variants alter TMW cell function.

ORIA GRANT

DR ANTHONY COOK, A/ PROF ALEX HEWITT

ASSESSING ABCA1 FUNCTION IN POAG USING PATIENT-SPECIFIC TRABECULAR MESHWORK CELLS

15Progress Report

A/Prof Alex Hewitt

PROGRESS REPORT


OVERVIEW

A significant barrier towards the evaluation of new therapies for glaucoma glaucoma1,2 is the lack of sensitive and precise measures of disease progression. As glaucoma is an optic neuropathy that results in the loss of retinal ganglion cells (RGCs), current measures of disease progression include a clinical examination of the appearance of the optic nerve for the loss of these cells, and visual field testing (or measurement of sensitivity to light throughout the field of vision) to capture the visual loss secondary to such cell loss. However, these measures are highly variable, meaning that the ability to accurately determine whether an individual is truly exhibiting glaucomatous progression or not is difficult. This is a tremendous barrier towards the evaluation of new therapies in glaucoma, and therefore our research sought to address this gap by developing highly precise measures of disease progression in glaucoma.

Fundus-tracked perimetry is a technique that could potentially provide a sensitive measure to monitor disease progression. Although this technique measures visual sensitivity in a similar way to conventional visual field testing, a key difference is that it utilises a high-resolution retinal camera to monitor the eye throughout the examination. Visual sensitivity can therefore be measured at specific locations in the retina, unaffected by eye movements that contribute to the high level of variability (even in vigilant test takers).3

In addition, the identical retinal location can be tested at subsequent visits, potentially enabling precise evaluation of disease progression over time. We therefore sought to determine the utility of this technique in eyes with glaucoma.

RESULTS

We embarked on a prospective study to evaluate the variability of visual field testing using fundus-tracked perimetry. Since measurements can be obtained precisely at specific retinal locations, this opened up the possibility of performing visual function testing at high density around the optic nerve head (seven times higher than conventional testing), where the earliest changes of glaucomatous visual function loss may be captured.

We observed that visual sensitivity measurements in areas of glaucomatous damage did not demonstrate high levels of variability as observed in previous studies,4 which were approximately twice to three times higher than what was observed in this study. This suggested that fundus-tracked perimetry could be used to as a precise tool to detect glaucoma progression (Wu et al, Invest Ophthalmol Vis Sci, 2016).

Although we had planned to evaluate how well this technique could detect glaucomatous progression over time, we identified another promising method for this purpose. Analogous to an electrocardiogram for the heart, the electroretinogram (ERG) is an objective measure of the functional state of the RGCs affected in glaucoma.

ORIA / EYE SURGEON’S FOUNDATION GRANT

DR ZHICHAO WU & PROF JONATHAN G. CROWSTON

USING FUNDUS-TRACKED PERIMETRY TO ENABLE PRECISE EVALUATION OF DISEASE PROGRESSION IN GLAUCOMA

17Progress Report

Using a novel simple-to-implement handheld ERG system, we developed a rapid testing protocol (requiring 1-minute per measurement) that reduced measurement variability by nearly 70% when compared to conventional methods (Wu et al, Trans Vis Sci Tech, 2016).

As a result of these findings, we now plan to begin a study to evaluate the effectiveness of both of these techniques combined for precisely monitoring glaucomatous progression over time.

References

1. Chang EE, Goldberg JL. Glaucoma 2.0: neuroprotection, neuroregeneration, neuroenhancement. Ophthalmology 2012;119:979-86.

2. Casson RJ, Chidlow G, Ebneter A, et al. Translational neuroprotection research in glaucoma: a review of definitions and principles. Clin Experiment Ophthalmol 2012;40:350-7.

3. Maddess T. The Influence of Sampling Errors on Test–Retest Variability in Perimetry. Invest Ophthalmol Vis Sci 2011;52:1014-22.

4. Wall M, Woodward KR, Doyle CK, Artes PH. Repeatability of automated perimetry: a comparison between standard automated perimetry with stimulus size III and V, matrix, and motion perimetry. Invest Ophthalmol Vis Sci 2009;50:974-9.

Publications arising from this ORIA funding, acknowledging its support

1. Wu Z, McKendrick AM, Hadoux X, Fan Gaskin, JC, Ang GS, Sarossy MG, Crowston JG. Test–Retest Variability of Fundus-Tracked Perimetry at the Peripapillary Region in Open Angle Glaucoma. Invest Ophthalmol Vis Sci, 2016;57:3619-25.

2. Wu Z, Hadoux X, Hui F, Sarossy MG, Crowston JG. Photopic Negative Response Obtained Using a Handheld Electroretinogram Device: Determining the Optimal Measure and Repeatability. Trans Vis Sci Tech, 2016;5:8

Dr Richard Wu

PROGRESS REPORT


OVERVIEW

The retinal photoreceptors form the first sensory element in the visual system. Recent advances in retinal imaging by incorporating of adaptive optics (AO) technology into clinical ophthalmic imaging devices enable visualisation of individual cone photoreceptor cells. To date, however, there are no consensuses on the optimal methods of adaptive optics image collection, post-acquisition image processing, nor on the selection of sampling windows for cone density measurements. Moreover, the limited information is available on the relationship between cone distribution and their function that can be measured using microperimetry, within central 20° of the retina.

AIM

The purpose of this pilot study was to:

i) optimise AO imaging protocol and develop software for reliable automated cone counting at specified locations of the macula,

ii) examine the relationship between retinal sensitivity and cone density in para- and peri-foveal regions in healthy subjects,

iii) assess the predictive value of cone density deviation from normative values in predicting retinal sensitivity loss in photoreceptor cell diseases.

RESULTS

In this study we used adaptive optics flood illumination ophthalmoscopy AO-FIO (rtx1, Imagine Eyes, Orsay, France) to image cone photoreceptors cells of 32 control subjects and 18 cases.

The cases comprised of 10 patients with scotoma (a first sign of macular disease) and 8 patients with suspected hydroxychloroquine (HCQ) toxicity after continuous HCQ dosing for 10 years. Quantitative analysis has been performed on adaptive optics images to measure the cone density at 36 microperimetry loci using a specialised software developed for this project. This software incorporates the circular Hough transform to detect cone reflexes and allows automated cone counting and calculation of cone density. The means of the retinal sensitivity and cone densities of control subjects at each of the loci location as well as illustrative cases of patients with the retinal diseases are presented in Figure.

Overall, in healthy subjects, the retinal sensitivity was the highest in the foveal region where the cone density riches the maximum. There was a reduction in sensitivity and cone density for all subjects with scotoma and patients with established retinal toxicity. A reduction in only cone density was observed in patients without established toxicity.

ORIA GRANT

DR DANUTA M BUKOWSKA AND DR EVAN WONG

STRUCTURE AND FUNCTION CORRELATION IN HUMAN MACULA – RELATIONSHIP BETWEEN CONE PACKING DENSITY AND RETINAL SENSITIVITY.

19Progress Report

IMPLICATIONS FOR PRACTISE/SCIENCE

The array of therapeutic options available to clinicians for treating retinal disease is growing. With these advances comes the need for non-invasive imaging of the retinal structures on a cellular level, to better monitor the efficiency of those therapies. In vivo cellular imaging of the human retina can be achieved by the incorporation of adaptive optics (AO) technology into clinical ophthalmic imaging devices. However, lack of consensuses procedures of AO image collection and processing, and limited information on the relationship between retinal sensitivity and cone distribution within the retina prevent ophthalmologists from using AO in their daily clinical practice.

This study has optimized image acquisition and developed novel analysis algorithms of cone photoreceptor cells images obtained using AO camera. Moreover, our pilot data presents the relationship between retinal sensitivity and cone density in para- and peri-foveal regions in healthy and diseased subjects. This is a first step towards providing a more sensitive measure of disease progression to support clinical trials to be conducted to demonstrate efficacy of novel treatments. However, future study with a larger sample of healthy and case subjects is required before this goal can be achieved.

Peer-reviewed publications arising from ORIA support for projects in the recent past

E. N. Wong, A. L. Chew, W. H. Morgan, P. J. Patel, F. K. Chen, “The use of microperimetry to detect functional progression in non-neovascular age-related macular degeneration: A systematic review,” Asia Pac J Ophthalmol 6(1), 70-79, 2017.

D. M. Sampson, D. Alonso-Caneiro, A. L. Chew, T. Lamey, T. McLaren, J. De Roach, and F. K. Chen, “Enhanced visualization of subtle outer retinal pathology by en face optical coherence tomography and correlation with multimodal imaging,” PloS One 11(12), 1-32, 2016.

D. M. Bukowska, S. L. Wan, A. L. Chew, E. Chelva, I. Tang, D. A. Mackey, and F. K. Chen, “Fundus autofluorescence in rubella retinopathy: correlation with photoreceptor structure and function,” Retina 37(1), 124-134, 2017.

Presentations on this work, acknowledging the ORIA

D. M. Sampson, “What can we learn from multimodal imaging of the human retina,” Optical Society 2016 lecture series, 7 December 2016, Queensland University of Technology – invited talk.

Figure 1. Graphical display of retinal sensitivity (top row) and cone density (bottom row) in the horizontal (left side) and vertical (right side) meridians. X-axis shows eccentricity in degrees from foveal center. Bars represent -1 to +1 standard deviation of the normative data. Blue dots represent microperimetry and cone density means for control subjects. Orange, green and red dots are example cases of scotoma, not-established and established retinal toxicity respectively.

Dr Danuta Bukowska

Dr Evan Wong

PROGRESS REPORT


OVERVIEW

The need for this project

Glaucoma is a leading cause of irreversible blindness which affects approximately 3% of the Australian population over 40 years of age and is often associated with increased ocular pressure. This disease is caused by the gradual loss of retinal ganglion cells (RGCs), the neurons which send visual information to the brain. The causes of glaucoma and the actual mechanisms underlying the loss of RGCs remain elusive, however, it is now believed that the initial cellular insults reside in the optic nerve head (ONH), where the RGC processes, or axons, pass out of the eye and into the optic nerve. We sought a better understanding of the cellular signalling processes occurring in this region, in particular involving mitogen-activated protein kinases (MAPKs), a large enzyme family known to have an involvement in neuronal death.

A note on our experimental approach

We used a laser to block fluid movements in the front of the rat eye. This mimics glaucoma by causing a sustained build-up of ocular pressure, which in turn exerts a damaging effect on the RGC axons in the back of the eye. By collecting tissues after specific periods of time, we could investigate cellular processes occurring as a result of pressure elevation, particularly relating to MAPKs.

RESULTS

MAPKs are broadly divided into three sub-groups: P42/44 MAPK, P38 MAPK and SAPK. We found that each of these enzyme groups were present in specific populations of cells throughout the untreated retina, ONH and optic nerve. After elevation of ocular pressure, alterations were observed:

• P42/44 MAPK was clearly activated in the ONH soon after elevation of pressure, in specific cells called astrocytes, which feed and pack around RGC axons.

• P38 MAPK became activated in microglial cells, throughout the tissue, after elevation of pressure. These cells are involved in immune system responses in nervous tissues. Interestingly, this enzyme was also activated in RGCs, themselves, in the retina.

• SAPK was present and activated throughout RGCs and their axons, even without increased ocular pressure. However, the pressure elevation caused this enzyme to accumulate in RGC axons passing through the ONH, ie. where damage initiation occurs.

ONGOING STUDIES

We are currently completing studies investigating compounds that block the action of each of the sub-types of MAPKs, to see whether these can prevent tissue damage after elevation of ocular pressure.

ORIA GRANT

DR JOHN P. M. WOOD

MITOGEN-ACTIVATED PROTEIN KINASES IN THE OPTIC NERVE HEAD IN EXPERIMENTAL GLAUCOMA

21Progress Report

IMPLICATIONS

We have shown that each of the three types of MAPK are activated in the ONH as a result of elevating pressure. Interestingly, the three types of MAPK are activated in three different cell-types. By investigating whether inhibitory compounds specific to each of the sub-types of MAPK have a positive effect on tissue outcome after induction of tissue injury, we can hypothesise as to which cell types and MAPK sub-classes are involved in RGC loss in glaucoma. Overall, the data obtained could lead to novel therapeutic targets for glaucoma and, ultimately, a reduction in the disease burden at the individual, population and economic levels.

Publications prepared from this study

• Mammone T, Chidlow G, Casson RJ, Wood JPM. Expression and activation of mitogen-activated protein kinases in the optic nerve head in a rat model of ocular hypertension. Submitted.

• Mammone T, Chidlow G, Casson RJ, Wood JPM. Improved immunohistochemical detection of phosphorylated MAP kinases in the injured rat optic nerve head. J Histochem Cytochem, submitted.

Figure 2. Localisation of active MAPK sub-types in ONH of rats subjected to experimentally elevated ocular pressure. Each of the three MAPK groups is represented: P42/44 MAPK, which is activated in astrocytes, SAPK (which is active in RGC axons), P38 MAPK, which is activated in microglial cells.

Dr John Wood

PROGRESS REPORT


OVERVIEW

Uveitis – inflammation inside the eye – is a cause of substantial visual morbidity that reduces quality of life and has a high economic cost. Based on calculations made with US data, the total annual cost of blindness from uveitis equals that of blindness from diabetic eye disease, although uveitis is far less common. Uveitis may represent an infection, or it may be a non-infectious autoimmune or autoinflammatory condition. For one in two patients with non-infectious uveitis, conventional immunosuppressive drugs will not adequately control the disease or treatment-limiting complications will develop. Quality of life in persons with non-infectious uveitis is considerably lower than would be expected based on level of vision alone. When the inflammation involves the posterior eye, risk of vision loss, reduction in quality of life and economic cost are highest. With the support of ORIA, we have explored a novel treatment paradigm for non-infectious posterior uveitis.

In a proof-of-concept study we investigated the effect of transcription factor blockade on induced or pathological – versus constitutive or homeostatic – expression of an endothelial adhesion molecule in a human uveitis model. Non-infectious posterior uveitis involves the migration of multiple leukocyte subsets across the retinal vascular endothelium.

Intercellular adhesion molecule-1 (ICAM-1) is a key adhesion molecule for leukocyte egress into human retina, and tumour necrosis factor alpha (TNF-α) is a master cytokine regulator of inflammation within human retina. We treated human retinal endothelial cells with TNF-α, and observed a substantial increase in ICAM-1 transcript expression, beginning within minutes of exposure. This finding indicated that ICAM-1 gene transcription might be targeted to limit induction of ICAM-1 protein expression on retinal endothelial cells and leukocyte-retinal endothelial interactions, without impacting consitutive levels of these parameters.

RESULTS

We observed an increase in the transcription factor, nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 (NF-κB1), in parallel with ICAM-1, in human retinal endothelial cells treated with TNF-α, and identified putative binding sites for NF-κB1 within the ICAM-1 regulatory region. We silenced NF-κB1 transcript with small interfering (si)RNA to examine the effect of targeting TNF-α-induced ICAM-1 gene transcription in human retinal endothelial cells. We measured the effect of NF-κB1 silencing on human retinal endothelial cell surface ICAM-1 expression, using a cellular ELISA. Knockdown of NF-κB1 significantly decreased TNF-α-induced ICAM-1 protein expression, but did not reduce constitutive ICAM-1 expression.

ORIA/G J WILLIAMS BEQUEST GRANT

PROF JUSTINE R. SMITH, FRANZCO, PHD, FAHMS

“ICAM-1 EXPRESSION IN RETINAL ENDOTHELIAL CELLS”

23Progress Report

We next examined the effect of NF-κB1 silencing on leukocyte-endothelial cell interactions, by quantifying THP-1 leukocyte binding to human retinal endothelial monolayers. Knockdown of NF-κB1 significantly decreased leukocyte binding to human retinal endothelial monolayers that were pre-treated with TNF-α, but did not reduce binding to non-treated monolayers.

In summary, NF-κB1-targeted blockade of ICAM-1 gene transcription reduces TNF-α-induced inflammatory activities of human retinal endothelial cells that contribute to the development of non-infectious posterior uveitis. Importantly, the blockade does not impact baseline levels of these activities. Retinal endothelial ICAM-1 transcription blockade has immense potential as a treatment for patients with non-infectious posterior uveitis. Importantly, this proof-of-concept research paved the path to a 3-year NHMRC Project Grant, for which funding commenced in 2017.

National grant funding that was obtained as a result of ORIA support:

National Health & Medical Research Council Project Grant

APP1123684: Regulation of ICAM-1 Expression in Human Retinal Endothelial Cells

Awarded to: Justine R. Smith (CIA), Flinders University, 2017-2019

Publication of this work that has acknowledged or will acknowledge ORIA support:

1. Ashander LM, Appukuttan B, Ma Y, Gardner-Stephen D, Smith JR. Targeting endothelial adhesion molecule transcription for treatment of inflammatory disease: a proof-of-concept study. Mediators Inflamm 2016; 2016: 7945848.

Two more publications related to this research are in draft form, with submission to peer-reviewed journals anticipated in 2017.

Prof Justine Smith and team at Flinders University, Adelaide

PROGRESS REPORT


OVERVIEW

The major obstacles in developing treatment for inherited retinal blindness are: (1) the large number of disease-causing genetic mutations, (2) the lack of suitable animal models for all mutations and (3) the inability to conduct clinical trials due to small number of individuals affected by each mutation. In the last decade, the advent of induced pluripotent stem cells (iPSC) has opened new avenues for translational research into human genetic diseases. Ten years ago (2006), it was shown that human skin cells could be reprogrammed into a cell that has similar properties to an embryonic stem cell, known as pluripotent state. These iPSC can be cultured to produce any cell type in the body, including cells found in the retina.

In this project, we aimed to produce iPSC from a patient with autosomal dominant retinitis pigmentosa caused by mutation of the RP1 gene. Mutations in this gene are associated with up to 10% of autosomal dominant retinitis pigmentosa. To establish an experimental platform for understanding RP1 disease mechanisms and evaluating novel treatments, we aimed to convert patient iPSC into retinal tissues so that we don’t have to remove patients’ retina for laboratory experiments. Since RP1 is only expressed in photoreceptors in the retina, this approach is essential to provide experimental access to tissues that would normally be unavailable for molecular analysis.

RESULTS

We have successfully reprogrammed patient skin cells into iPSCs. Patient iPSC expressed markers of embryonic stem cells and could be differentiated to produce beating heart muscle cells and various forms of retinal cells in our laboratory. Overall, patient iPSC were indistinguishable from iPSC derived from healthy volunteers. To our knowledge, our RP1-iPSC line, funded by ORIA grant, was the first iPSC line produced from a patient in Western Australia. Establishment and characterization of this first iPSC line required numerous attempts using different reagents and protocols. However this arduous work of trying out different methods has resulted in an optimized protocol that allows us to generate high quality iPSC lines quickly and efficiently. Since the establishment of capability to produce iPSC at the Lions Eye Institute, we were able to collaborate with several other groups in Western Australia to study other non-ocular conditions using iPSC as a disease model (see below).

Patient and control iPSC were cultured to produce miniature retina in the petri dish known as retinal organoids. We confirmed the presence of rod and cone photoreceptors, Muller cells and retinal ganglion cells in these retinal organoids. Retinal differentiation into organoid was comparable between patient and control iPSC cultures. Molecular characterization of patient and control retinal organoids is currently underway to investigate subtle differences between patient and healthy organoids so that we can identify markers that represent the earliest sign of retinitis pigmentosa developing in these organoids.

ORIA/ESME ANDERSON GRANT

DR SAMUEL MCLENACHAN AND DR FRED CHEN

DEVELOPING PERSONALISED DISEASE MODELING FOR TESTING NOVEL TREATMENT IN INHERITED RETINAL DISEASE DUE TO RP1 MUTATION

25Progress Report

With the initial delay in iPSC reprogramming and the long time course of retinal differentiation protocols (3-6 months), we anticipate completion of Aims 2-3 in early 2018.

IMPLICATIONS

Molecular characterisation of our RP1-iPSC-derived retinal cells is expected provide valuable insights into the mechanisms where by mutant RP1 proteins causes disease as well as invaluable data on the efficacy of new therapeutic strategies, such as RNA interference and translational read-through inducing drugs for the treatment of this disease. In a broader context, the successful reprogramming of the RP1-iPSC line was an essential first step in bringing iPSC technology to Western Australia. Pilot data generated from this project has stimulated significant interest from local and international researchers. To date, we have generated iPSC disease models for six inherited retinal diseases, establishing collaborations with Murdoch University, Telethon Kids Institute, Ear Science Institute and the University of Skovde. In summary, we have added a valuable new capacity for personalised disease modeling research in Western Australia, paving the way for future discoveries and ultimately, new treatments to prevent blindness due retinal dystrophy.

Figure 3. RP1-iPSC-derived Retinal Organoids

Dr Sam McLenachan and Dr Fred Chen

PROGRESS REPORT


OVERVIEW

Age-Related Macular Degeneration (AMD) is the leading cause of blindness in the developed world, costing the Australian economy alone up to $5 billion per annum. The need for a therapeutic is paramount, yet the most common subtype of the disease, known as atrophic or ‘dry’ AMD, has no effective therapies. MicroRNA (miRNA) molecules have gained traction as potential gene regulatory therapeutics due to their unique ability to target multiple genes working within the same cellular pathway, effectively getting to the root of the problem. The eye is an excellent neuronal model for investigating the endogenous functions of miRNA and their role in neurodegeneration, first because miRNAs are abundant in the central nervous system including the retina, and second because the eye is a closed compartment, where constructs can be easily delivered without interfering with the target tissue, and without the complications of surgery. This project seeks to understand two microRNA molecules, miR-124 (anti-inflammatory) and miR-155 (pro-inflammatory) and their potential as therapeutics for retinal degenerations, including AMD.

RESULTS

We have demonstrated that an absence of miR-124 in human AMD retina is correlated to photoreceptor degeneration (Figure 1A-C). This also holds true in our mouse model of photo-oxidative damage (PD), which mimics facets of atrophic AMD, which indicates a change in expression levels of miR-124 after retinal damage is incurred

(Figure 1D-J). Further we found that modulation of miR-124 has the capacity to control the inflammatory environment of the retina, by regulating the expression of pro-inflammatory chemokine CCL2, which we have previously shown to be involved in the progression of retinal degenerations (1, 2). Increasing availability of miR-124, with the use of therapeutically delivered mimics, reduced photoreceptor cell death, decreased macrophage recruitment into the outer nuclear layer, and increased retinal function in the degenerating retina (Figure 1K-L). We also localised the expression of miR-124 in the degenerating retina to be in both neuronal and Müller cells. For miR-155, we have demonstrated that it directly targets complement factor H, a main player in AMD (3). Furthermore, we found that intravitreal delivery of miR-155 inhibitors also increased retinal function (Figure 1M-N).

Overall, our results help to further elucidate the role of miR-124 in the retina and provide a solid foundation for further study on the topic. Taken together, our results have strongly demonstrated the potential of miR-124 mimics and miR-155 inhibitors as a combinational gene therapy in retinal degenerations. This further paves the way for investigations exploring the therapeutic potential of miR-124 and miR-155 in diseases where inflammation is a key feature of pathogenesis, including AMD, Parkinson’s disease and Alzheimer’s disease.

We would like to thank the Ophthalmic Research Institute of Australia (ORIA) and the Eye Surgeons’ Foundation (ESF) for the support of this work.

ORIA/EYE SURGEONS’ FOUNDATION GRANT

DR R NATOLI, A/PROF R ESSEX & PROF J PROVIS

MICRORNA PROVIDE A NEW CLASS OF THERAPEUTICS FOR DISEASES SUCH AS AGE-RELATED MACULAR DEGENERATION

27Progress Report

As a direct result of this funding we were able to obtain ground-breaking preliminary data which enabled us to secure National Health and Medical Research Funding (The use of microRNA as novel therapeutic targets for reducing retinal inflammation and degeneration. NHMRC: 1127705; 2017-2019). Further, these results will form the basis for two high impact publications. We would like to thank both funding agencies for supporting Early Career Researchers, innovative science and our quest to slow the progression of retinal degenerations.

Publications arising from this project:

Due to increased funding from the NHMRC we have expanded the scope of this project which has delayed publication. The first publication to arise from this work is due to be submitted in May, 2017, with the remaining work to be published before the end of 2017. Both ORIA and ESF will be acknowledged in these publications.

References:

1 .M. Rutar, R. Natoli, J. M. Provis, Small interfering RNA-mediated suppression of Ccl2 in Muller cells attenuates microglial recruitment and photoreceptor death following retinal degeneration. J Neuroinflammation 9, 221 (2012).

2. M. Rutar, R. Natoli, K. Valter, J. M. Provis, Early focal expression of the chemokine Ccl2 by Muller cells during exposure to damage-inducing bright continuous light. Invest Ophthalmol Vis Sci 52, 2379-2388 (2011).

3. G. S. Hageman et al., A common haplotype in the complement regulatory gene factor H (HF1/CFH) predisposes individuals to age-related macular degeneration. Proc Natl Acad Sci USA 102, 7227-7232 (2005).

Figure 4. (A) In situ hybridization of miR-124 in normal age-matched human retina shows strong staining of in the INL and outer limiting membrane compared with (B), an AMD affected retina where staining of miR-124 is depleted. (C) The scrambled control probe showed no labelling in the human retina. (D-E) The difference in distribution of miR-124 in control compared to photo-oxidative damaged (PD) mice is shown in whole retinal sections.

The central regions of the superior side of the retina (rectangles in D and E) show the location for greatest susceptibility to cell death. (F-I) Control animals show labelling predominantly below the ONL in the OLM, while damaged retinas display higher amounts in the INL and depletion of labelling in the OLM comparatively.This change is seen as early as day 1, with consistent labelling at day 3 and day 5 of PD. (J)

Scrambled control probe shows no retinal staining. (K-L) Electoretinography (ERG) showed a decreased response amplitude in PD mice receiving a control, scrambled miRNA compared with those injected with miR-124 mimic in the a-wave and b-wave. (M-N) Intravitreal delivery of miR-155 inhibitor revealed no significant change in ERG a-wave but a significant increase in the b-wave (*indicates significance using a Student’s t-test or two-way ANOVA with Sidak’s post hoc test for multiple comparisons, P < 0.05 error bars indicate SEM)

Dr Ricardo Natoli

PROGRESS REPORT


OVERVIEW

Why was there a need for this project?

Glaucoma is typically understood to be an intraocular pressure (IOP)-associated optic neuropathy and almost all treatment options in glaucoma involve the lowering of IOP. The disease process causes death of the retinal ganglion cells (RGCs); it is the axon fibres of these cells that make up the bulk of the optic nerve. In clinical and experimental models of glaucoma, RGCs can be conceptualised into three groups: healthy, dead, and “sick” ganglion cells.1 The conventional, established methods of monitoring glaucoma were not designed to assess the state of these “sick” cells but only examine the structural and functional outcomes following ganglion cell death, and their greatest limitation is that it can takes months, if not years, to detect worsening of disease.

Glucose eye drops have recently been demonstrated to temporarily improve vision in the form of acuity and contrast sensitivity in patients with primary open-angle glaucoma (POAG).2 This is the first time that short-term improvement in visual function has been demonstrated without the lowering of IOP in POAG. It was postulated that vision improvement occurred as a result of glucose providing an alternative energy supply to metabolically challenged RGCs. The exact site in the retina, both in terms of the layer and topographical location at which this improvement is mediated, was explored in this study, thereby elucidating the underlying mechanism that facilitates this improvement.

We investigated subjects before and after administration of topical glucose with two technologies:

1) The photopic negative response (PhNR) of the full-field electroretinogram, which has been demonstrated to be an accurate and sensitive means of detecting change in RGC function on glaucoma.

2) Microperimetry for monitoring short-term functional change at retinal locations at highest risk of change.

RESULTS

Electroretinogram (ERG) recordings were performed before and after topical glucose drops using a hand-held ERG device (RETeval system, LKC Technologies, Inc., Gaithersburg, MD, USA). Artefacts such as blinks were removed after the recording and different processing methods were applied to optimise the signal and obtain an adequate baseline correction for the average response. To date we have found no significant group difference in PhNR before and after glucose treatment.

We have completed the microperimetry component of our study, and surprisingly a decline in the overall sensitivity was demonstrated following the administration of topical glucose, rather than an improvement (see Figure 1).

ORIA NEW INVESTIGATOR GRANT

DR JENNIFER FAN GASKIN

HOW DOES GLUCOSE BOOST VISION?

29Progress Report

What are the implications for clinical practice?

Whilst we cannot firmly conclude that topical glucose does not temporarily improve visual function, neither the PhNR nor our method of recording or microperimetry has proven to be a useful tool in detecting this improvement. One possible reason for a failure to detect a change in the PhNR is that the majority of subjects involved in this study had moderate to advanced glaucoma, and the potential neuroprotective effect of glucose may be limited in this group.

The reason for the decline in post-glucose sensitivity in microperimetry may be due to fatigue of the subjects on repeat testing, as this was also exhibited in a separate study testing a different topographic region in glaucoma patients.3 More selective subject recruitment and refinement of testing protocol may help definitively determine whether the PhNR and microperimetry are useful tools for detecting improvement in glaucoma following glucose administration.

References

1. Casson RJ, Chidlow G, Ebneter A, et al. Translational neuroprotection research in glaucoma: a review of definitions and principles. Clin Experiment Ophthalmol 2012;40: 350–7

2. Casson RJ, Han G, Ebneter A, et al. Glucose-induced temporary visual recovery in primary open angle glaucoma. Ophthalmology 2014;121:1203-11

3. Wu Z, McKendrick AM, Hadoux X, Fan Gaskin JC, Ang GS, Sarossy MG, Crowston JG. Test-Retest Variability of Fundus-Tracked Perimetry at the Peripapillary Region in Open Angle Glaucoma. Invest Ophthalmol Vis Sci 2016;57:3619-25

Figure 5. A significant decline in mean point-wise sensitivity of the central 10° visual field was demonstrated on microperimetry following the administration of topical glucose eye drops (mean point-wise sensitivity 25.0 dB pre-glucose vs. 24.4 dB post glucose, p=0.002. n= 7).

Dr Jennifer Fan Gaskin

PROGRESS REPORT


OVERVIEW

Retinal Muller glial cells, the major supporting cells in the retina, play an important role in maintaining normal retinal homeostasis. They particularly participate in providing metabolic support to photoreceptors. Dysfunction of Muller cells has been implicated in some retinal diseases such as Macular Telangiectasia type 2 (MacTel2), which is an untreatable macular disease with unknown cause. Despite the importance of Muller cell function, the method to generate Muller cells from human pluripotent stem cells is yet to be discovered.

The main aim of the project is to develop a method to make Muller cells from human pluripotent stem cells including embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) from MacTel2 patients and healthy subject. We particularly modulate the Notch signalling pathway in retinal progenitor cells derived from the stem cells. The Notch signalling pathway is a highly conserved pathway in the central and peripheral nervous systems and heavily involved in determining cell fate such as self-renewal and differentiation. It plays an important role in glial cell differentiation including retinal Muller cells from retinal progenitor cells.

RESULTS

For the past year, we have been successful to generate retinal Muller glial cells from human pluripotent stem cells, by promoting the Notch signalling pathway in retinal progenitors. We firstly used hESCs to develop the method. We differentiated hESCs into retinal progenitor cells and promote the Notch signalling pathway with the Notch ligands, DLL4 and Jagged-1 (50ng/ml) for 2-6 weeks. We validated Muller cell differentiation with 4 different methods including qRT-PCR, immunocytochemistry, western blot and fluorescence activated cell sorting, all of which showed upregulation of the most distinctive Muller cell marker, Glial Fibrillary Acidic Protein (GFAP) (Figure1). The results provided a solid foundation for the next step of the project which includes generation of Muller glial cells from human iPSCs and pre-clinical transplantation study of grafting Muller precursor cells-derived from stem cells into a transgenic mouse retina with Muller cell loss.

We have been successful in making iPSCs from two of MacTel2 patients’ skin biopsies in collaboration with A/P Alice Pebay’s lab at Centre for Eye Research Australia. We recently used these iPSCs and the iPSCs from healthy subject to generate Muller cells from their retinal progenitors. We used the method described above, and GFAP positive cells were found from immunnohistochemstry after 4-6 weeks of Notch promotion (Figure 1, D and E). We are currently in the process of validating the results.

ORIA NEW INVESTIGATOR /RANZCO NSW BRANCH GRANT

DR S H CHUNG & PROF M GILLIES

USE PATIENT-DERIVED STEM CELLS TO MODEL MACULAR TELAGIECATASIA TYPE 2 IN A DISH

31Progress Report

RESEARCH OUTPUT

With generous support from ORIA toward to the project in 2015, we were able to apply for an Australian provisional application with the method to differentiate Muller cells from human pluripotent stem cells in the past year, and produced one conference abstract at the International Society of Stem cell Research 2016 in San Francisco. We are also in the process of writing a manuscript.

Figure 6. (A) A schematic diagram of Muller cell differentiation method from human stem cells. (B-D) Immunocytochemisty confirmed GFAP positive cells after 2-6 weeks after Notch promotion. B and C represent Muller cells derived from hESC, and D and E are Muller cells derived from iPSCs (both MacTel2 and healthy subject). (F and G) qRT-PCR analyses revealed a significant upregulation of GFAP mRNA transcription. (H and I) FACS quantification of GFAP positive cells after 4 weeks (H) and 6 weeks (I) Notch promotion on hESC derived retinal progenitors. (J) Western blot indicates upregulation of GFAP protein in the Notch ligand treated retinal progenitors. * represents p< 0.05. Scale bar =100µm

PROGRESS REPORT


ORIA NEW INVESTIGATOR GRANT

NUCLEAR ESCAPE ROUTE FROM VISION LOSS IN LEBER HEREDITARY OPTIC NEUROPATHY

OVERVIEW

LHON is a devastating disease that affects predominantly young men and can cause sudden and irreversible blindness. LHON is maternally-inherited and causes dysfunction in the powerhouse of cells, mitochondria. For unknown reasons, within the same family, a sibling might go blind because of LHON (LHON-affected), whilst other siblings, although identical from a mitochondrial genetics point of view, may remain asymptomatic and maintain good vision throughout life (LHON-unaffected).

My work investigated whether cells from LHON-unaffected people compensated for the mitochondrial defect caused by LHON through the activation of other bioenergetic pathways. I answered this question definitely for the first time in a tightly controlled cohort, showing that contrary to some previously published findings, cells from LHON-unaffected people do not exhibit bioenergetic compensation. Instead, I found evidence of subtle, alternative compensatory mechanisms in asymptomatic LHON patients that requires further investigation. These results are currently being prepared for publication.

RESULTS

Understanding why some people who carry genetic LHON defects lose vision whilst others preserve good vision throughout life will lead to effective therapeutic targets being identified. This work has answered the first question on this path, establishing that there is no bioenergetic compensation in LHON. This is crucial knowledge, as most therapeutic approaches tested in LHON have so far focused on increasing bioenergetic metabolism. Unfortunately, these therapeutic approaches have resulted in scarce clinical benefit.

I am very grateful to ORIA for the New Investigator Grant I was awarded in 2016. The ORIA funds were used to support research - chemicals, consumables and partial salary funding, into the genetic pathways that mediate protection in Leber’s hereditary optic neuropathy (LHON).

Thanks to the ORIA New Investigator Grant, I have generated sufficient preliminary data to request research funding from the United Mitochondrial Disease Foundation, to continue my investigation into the alternative cellular compensatory mechanisms that may prevent blindness in people at risk due to LHON.

DR I LOPEZ SANCHEZ

33Progress Report

Dr Isobel Lopez Sanchez

PROGRESS REPORT


PROJECT BRIEF BACKGROUND AND OBJECTIVES

Glaucoma is a common adult-onset neurodegenerative disease whose pathogenesis is still under intense debate. Glaucoma traditionally has been explained by increased intraocular pressure (IOP), however up to 50% of cases are not associated with elevated IOP. We are exploring other factors which may damage the optic nerve and contribute to glaucoma pathogenesis. Substantial evidence indicates that mitochondria within the retinal ganglion cells of the optic nerve suffer damage during glaucoma which impairs function of the optic nerve. Mitochondria are the powerhouses of the cell, providing the energy for the optic nerve cells to transmit signals from the eye to the brain. Using glaucoma patient cell lines, we are examining the protein machinery within the mitochondria (the oxidative phosphorylation pathway – OXPHOS) which is responsible for generating energy for the optic nerve to function. In previous ORIA-supported research we identified that a particular component of the protein machinery (Complex-I) is impaired in glaucoma, and results in less energy being generated.

In this current proposal, we propose to identify at the protein level if there are particular structural abnormalities which may explain the Complex-I failure. This was to be achieved by mass spectrometry-based analysis of the mitochondrial proteome with a particular focus on the candidate enzyme called Complex-I.

PROJECT PROGRESS

The first part of the project involved the isolation of high quality mitochondrial preparations. This was completed successfully by mid-2016. For this, 15 control lymphoblast cell lines and 15 primary open-angle glaucoma (POAG) lymphoblast cell lines were expanded to 500 ml cultures and mitochondria isolated using established protocols from the Trounce laboratory. In this process we chose to vary our aim to include in the downstream mass-spec analysis samples of ‘crude’ mitochondrial purity, together with Percoll-gradient purified samples. This will give extra information on proteins that associate closely with mitochondria from the so-called ‘mitochondrial-associated-membranes, or MAM, of the endoplasmic reticulum. All 30 cell lines were processed successfully, and the resulting mitochondrial fractions were sent to Co-Investigator Gruz’s lab in late 2016.

ORIA/RENENSSON BEQUEST GRANT

DR NICOLE VAN BERGEN, A/PROF IAN TROUNCE & PROF FRANK GRUS

MITOCHONDRIAL PROTEOMICS AS A SELECTIVE TOOL FOR UNRAVELLING POAG DISEASE PATHOGENESIS

35Progress Report

Proteomic analysis is ongoing in Prof. Gruz’s lab, with all indications showing excellent sample purity and consistency. The figure above shows an example of the purified mitochondrial proteins run on a denaturing gel before the indicated bands are excised and the proteins in each band identified using mass spectrometry. This work is currently underway and we expect to have a high impact publication from this, but more importantly we hope to identify new therapeutic targets involving OXPHOS complex I.

While we are disappointed to not have final findings to report, we are confident that high quality new information will be obtained in the second half of 2017 from this pioneering analysis of the mitochondrial proteome in glaucoma.

Dr Nicole Van Bergen

A/Prof Ian Trounce

PROGRESS REPORT


OVERVIEW

Brain derived neurotrophic factor (BDNF) has been shown to play a critical role in the preservation of inner retinal function and structural integrity in glaucoma. Our research has shown that BDNF/TrkB signalling regulates the glycogen synthase kinase 3 β (GSK3β) signalling in the retinal ganglion cells (RGCs) and other neuronal cells. Shp2 is a partner phosphatase for regulating the TrkB receptor signalling in the RGCs. Using a series of bioinformatics approaches, we have been able to determine the molecular constraints that determine the interaction of TrkB agonists with the TrkB receptor, thus enhancing our understanding of the TrkB actions. The regulatory phosphatase, Shp2 undergoes activation under high intraocular pressure (IOP) conditions leading to deactivation of the TrkB receptor. Activation of Shp2 is mediated through its interactions with the integral membrane protein caveolin. Recent studies have established variations in the caveolin gene locus as a strong risk factor for development of glaucoma.

This study proposed to investigate the involvement of Shp2 phosphatase and caveolin proteins in glaucoma and evaluate the effects of modulating Shp2 on the inner retina in glaucomatous conditions. Inner retina is particularly affected in glaucoma and so strategies that have protective effects on the inner retina may be useful in glaucoma. Interestingly, we have recently observed that transgenic rodent model of Alzheimer’s disease depicts inner retinal degenerative changes which will

help us to understand the molecular mechanisms underlying inner retinal degenerative processes. We have carried out studies in caveolin -1 knockout mice that we have obtained from our collaborator based in University of Queensland (Prof. Rob Parton). These animals were re-derived and a new colony established at Macquarie University. We have been able to design and obtain the adeno-associated vectors (AAVs) tagged with the green fluorescent proteins (GFPs). We have already been carrying out the AAV –transgene intravitreal injections into the rats and caveolin-1 knockout, heterozygous and wild type mice and have obtained good expression of Shp2 transgene. We have also carried out several control experiments in order to help us evaluate the effects of over-expression of these transgenes. We have performed both electrophysiological and structural studies to determine the effect of the gene therapy experiments.

RESULTS

Our results indicate that caveolin plays an important role in the preservation of inner retinal function as demonstrated by changes in the scotopic threshold recordings in these mice. Shp2 over expression in the RGCs reduced the amplitudes of electrophysiological recordings indicating compromised retinal function. Thickness of the ganglion cell layer was also reduced indicating that these cells are negatively affected by SHp2 over-expression. On the other hand our studies show that knockdown of Shp2 using AAV under the experimental glaucoma conditions was

ORIA/MARY TILDEN BEQUEST GRANT (2015)

DR VIVEK GUPTA, PROF STUART GRAHAM & DR YUYI YOU

DEVELOPING A NEW GENE THERAPY APPROACH TO PROTECT AGAINST GLAUCOMA DAMAGE

37Progress Report

protective for the retina as measured by electrophysiological recordings and histological analysis of the retina. These studies identify Shp2 as a novel molecule that can be targeted using genetic and pharmacological approaches for future drug development in glaucoma.

Publications:

N Chitranshi, Y Dheer, R Vander Wall, V Gupta, M Abbasi, SL Graham, Vivek Gupta (2016) Computational analysis unravels novel destructive single nucleotide polymorphisms in the non-synonymous region of human caveolin gene. Gene Reports. doi.org/10.1016/j.genrep.2016.08.008.

Gupta Vivek, Chitranshi N, You Y, Gupta V, Klistorner A, Graham S. (2014) Brain derived neurotrophic factor is involved in the regulation of glycogen synthase kinase 3β (GSK3β) signalling. Biochem Biophys Res Commun. 454(3):381-386.

Gupta V, Gupta VB, Chitranshi N, Gangoda S, Vander Wall R, Abbasi M, Golzan M, Dheer Y, Shah T, Avolio A, Chung R, Martins R, Graham S. (2016). One protein, multiple pathologies: multifaceted involvement of amyloid β in neurodegenerative disorders of the brain and retina. Cellular and Molecular Life Sciences 10.1007/s00018-016-2295-x.

Yuyi You, Vivek K Gupta, Nitin Chitranshi, B Reedman, A Klistorner, Stuart L Graham. (2015) Visual evoked potential recording

in a rat model of experimental optic nerve demyelination. Journal of Visualized Experiments. JoVE52934R3.

Chitranshi N, Gupta V, Kumar S, Graham SL (2015) Exploring the Molecular Interactions of 7,8-Dihydroxyflavone and Its Derivatives with TrkB and VEGFR2 Proteins. Int J Mol Sci. 16(9):21087-108. doi: 10.3390/ijms160921087.

Vivek K Gupta, Nitin Chitranshi, Veer B Gupta, Mojtaba Golzan, Yogita Dheer, Roshana Vander Wall, Dana Georgevsky, Anna E King, James C Vickers, Roger Chung, Stuart Graham (2016) Amyloid β accumulation and Inner retinal degenerative changes in Alzheimer’s disease transgenic mouse. Neuroscience letters, doi: 10.1016/j.neulet.2016.04.059.

Nitin Chitranshi, Vivek Gupta, Yogita Dheer, Veer Gupta, Roshana Vander Wall, Stuart Graham (2016) Molecular determinants and interaction data of cyclic peptide inhibitor with the extracellular domain of TrkB receptor. Data in Brief (2016) 776–782.

N Chitranshi, Y Dheer, RVander Wall, VGupta, Yuyi You, Alexander Klistorner, Stuart Graham, Vivek Gupta (2016) PTPN11 influences TrkB phosphorylation and endoplasmic stress response disrupting retinal integrity. (In preparation)

Oral / Poster presentations:

Stuart L. Graham, Nitin Chitranshi, Roger Chung, Vivek K Gupta (2015) Impaired inner retinal function and Amyloid β aggregation is demonstrated in a mouse model of

Alzheimer’s disease. Annual conference of Association for Research in Vision and Opthalmology (ARVO) , Colorado, USA.

Nitin Chitranshi, Yuyi You, Vivek K Gupta, A Klistorner, Stuart L. Graham (2015) Effects of isoflurane on the visual evoked potentials in rats. Annual conference of Association for Research in Vision and Opthalmology (ARVO), Colorado, USA.

Nitin Chitranshi, Vivek K Gupta, Roshana Vander Wall, Yogita Dheer, Stuart L Graham (2016) AAV mediated gene therapy to modulate neurotropic factors in the retina and in neuronal cells in culture. American society for Gene and cell Therapy, 16th annual meeting, Washington DC, USA, May 4-7, 2016.

Dheer, Yogita; Chitranshi, Nitin; Vanderwall, Roshana; Graham, Stuart L.; Gupta, Vivek (2016). Retinoid x receptor (RXR) expression in the rodent retina and effects of its modulation in neuronal cells. ARVO, 2016, May1-5, Seattle, USA.

Nitin Chitranshi, Vivek K Gupt, Roshana Vander Wall, Yogita Dheer, Stuart L Graham (2016) SHP2 (PTPN11) over-expression by AAV gene delivery impairs neuronal cell growth in SH-SY5Y cells and induces neurodegeneration of SD rat retinal ganglion cells. ARVO, 2016, May1-5, Seattle, USA.

Impaired Protease Regulatory Activity of Neuroserpin is Observed in Human Glaucoma Eyes and a Rodent Model of Experimental Glaucoma. Vivek Gupta, Roshana Vander Wall, Stuart Graham. Clin Exp Opthalmol 43, S1, 22.

Left to Right: Prof Stuart Graham, Dr Vivek Gupta and Dr Yuyi You

PROGRESS REPORT


OVERVIEW

Glaucoma remains one of the 3 major causes of blind registrations in Australia. High blood pressure (BP) has been regarded as a possible risk factor for the development and progression of glaucoma, but it has been postulated that it may be protective early in the disease process. Blood vessel abnormalities secondary to high BP could impair blood flow to the eye and initiate glaucomatous damage. On the other hand hypotension and reduced ocular perfusion pressure is known to be a risk factor as well. To further elucidate the role of high BP in glaucoma, we studied retinal vascular characteristics and structural changes in glaucoma and

hypertensive animal models.

RESULTS

We induced glaucoma using micro-bead injections in one eye of normal and hypertensive rats. The fellow eye served as control. At 12 weeks follow-up, mean eye pressure in the glaucoma and control eyes of the hypertensive rats was 22.7 ± 4.4 mmHg and 11.6 ± 1.0 mmHg, respectively. These values in the control rats were 21.5 ± 2.2 mmHg and 11.6 ± 1.4 mmHg.

Vascular Findings

The average static venous diameter was smaller in hypertensive rats, however, the difference did not register as statistically significant (P = 0.08).

The average arterial diameter was significantly narrower in hypertensive rats (P < 0.05). This was in line with previous studies which reported retinal arteriolar narrowing in hypertension. This phenomenon can be explained by increased vascular resistance due to vasoconstriction in response to high BP.

The average venous pulse amplitude was non-significantly lower in SHR rats (p = 0.08). However, the average arterial pulse amplitude was significantly smaller in this group (p <0.05). As expected, we observed pulse amplitude attenuation along arteries. In contrast, the pulse amplitude did not change along veins.

Structural Findings

In collaboration with University of California San-Diego, we employed a custom written algorithm to automatically segment and measure the retinal ganglion cell layer thickness (RGCLT) in each eye obtained using non-invasive optical coherence tomography scans.

While there was a reduction in the mean RGCLT in the glaucoma eyes of the hypertensive rats compared to their fellow eyes this did not reach significance (48.5 ± 7.4 μm and 62.5 ± 17.5 μm, respectively; p = 0.052). However, significant differences were found between the RGCLT thickness in the glaucoma and control eyes of the normal rats (43.3 ± 14.3 μm and 74.6 ± 16.5 μm, respectively; p < 0.001).

ORIA/RANZCO NEW INVESTIGATOR GRANT (2015)

DR MOJTABA GOLZAN & PROF ALBERTO AVOLIO

EFFECTS OF HIGH BLOOD PRESSURE ON RETINAL HEMODYNAMICS IN GLAUCOMA

39Progress Report

IMPLICATIONS

This study confirmed a narrower arterial diameter and pulse amplitude in hypertensive animals after 12 weeks. These changes were non-significant in venous diameter and pulse amplitudes. We also observed a non-significant loss of RGCLT in the eyes of hypertensive rats with experimentally induced glaucoma. In other words, hypertensive animals had less structural damage compared to normal eyes after glaucoma was induced.

Collectively, our findings are consistent with the concept that early in glaucoma high BP could be protective against a raised eye pressure, but that later, progressive blood vessel changes lead to an exacerbation in disease pathology.

Publications related

Rezaeian M, Georgevsky D, Golzan SM, Graham SL, “High speed in-vivo imaging of retinal hemodynamics in a rodent model of hypertension” IEEE Eng Med Biol, 2016, doi: 10.1109/EMBC.2016.7591420.

FINANCIALS

FINANCIALS


In accordance with a resolution of the directors, the directors submit herewith the financial statements of The Ophthalmic Research Institute of Australia for the year ended on 30 June 2017 and report as follows:

1. DIRECTORSThe names of the Directors of the company in office at the date of this report are:

Clinical Professor Stephanie Watson, Sydney (Chairman)

Professor Mark Gillies, Sydney (Vice Chairman)

Professor Richard Mills, Adelaide (Honorary Secretary)

Associate Professor Paul Healey, Sydney (Honorary Treasurer)

Dr Fred Chen, Perth

Professor Jonathon Crowston, Melbourne



Professor Stuart Graham, Sydney


Professor David Mackey, Perth

Professor Paul Mitchell, Sydney

Professor Peter J McCluskey, Sydney


Associate Professor Andrea Vincent, New Zealand

2. INFORMATION ON DIRECTORS

The names, qualifications and period membership commenced and position held are as follows:

Dr Fred Chen MB BS (Hons), PhD (London), FRANZCO, CSA (Cert) 2011

Professor J Crowston BSc, MB BS, FRCOphth, FRANZCO, PhD 2008

Dr Clare Fraser, MB BS, MMed, FRANZCO 2016

Dr Jennifer Fan Gaskin MBChB, MD, FRANZCO 2016

Professor Mark Gillies MB BS, PhD, FRANZCO Vice Chairman 2004

Professor Stuart L Graham MB BS, MS, FRANZCO, FRACS 2001

Associate Professor Paul Healey MB BS (Hons), B (Med) Sc,

MMed PhD, FRANZCO Honorary Treasurer 2011

Dr Wilson Heriot MB BS, FRANZCO, FRACS (Resigned 20 November 2016) 2009

Dr Gerald Liew, MB BS, MMed, PhD, FRANZCO 2015

Professor David Mackey MB BS, MD, FRANZCO, FRACS 2005

Professor Paul Mitchell MB BS, MD, PhD, FRANZCO, FRACS, FRCOphth, FAFPHM 2015

Professor Peter J McCluskey MB BS, FRANZCO, FRACS 1984

Dr John Males MB BS, M Med, FRANZCO 2009

Dr Richard Mills MB BS, FRCS, FRACS, FRANZCO, PhD Honorary Secretary 2003

Associate Professor Andrea Vincent MBChB, FRANZCO 2008

Clinical Professor Stephanie Watson BSc, MB BS, FRANZCO, PhD Chair 2006

Dr Andrew White BMedSci (Hons), MB BS, PhD, FRANZCO 2015

No shares are held by Directors.

43Financials

3. MEETINGS OF DIRECTORS

During the financial year three meetings of directors were held. Attendances were:

Board Members Number Eligible to Attend Number Attended

Dr Fred Chen, Perth 3 3

Professor J Crowston, Melbourne 3 3

Professor Mark Gillies, Sydney 3 3

Professor Stuart L Graham, Sydney 3 3

Associate Professor Paul Healey, Sydney 3 3

Dr Wilson Heriot, Melbourne 2 2

Dr Gerald Liew, Sydney 3 1

Professor David Mackey, Perth 3 3

Professor Peter J McCluskey, Sydney 3 2

Professor Paul Mitchell, Sydney 3 0

Dr John Males, Sydney 3 3

Proessor Richard Mills, Adelaide 3 3

Associate Professor Andrea Vincent, New Zealand 3 3

Clinical Professor Stephanie Watson, Sydney 3 3

Dr Andrew White, Sydney 3 1

Dr Andrew White, Sydney 3 1

4. INDEMNIFYING OFFICER OR AUDITORThe company has not during or since the financial year in respect of any person who is or has been an officer or auditor of the company or a related body corporate indemnified or made any relevant agreement for indemnifying against a liability incurred as an officer including costs and expenses in successfully defending legal proceedings or paid or agreed to pay a premium in respect of a contract of insurance against a liability incurred as an officer for the costs or expenses to defend legal proceedings.

5. PRINCIPAL ACTIVITIESThe principal activity of the company in the course of the financial period was to provide funds for ophthalmic research. There has been no significant change in the nature of this activity during that period.

6. SHORT-TERM AND LONG-TERM OBJECTIVESThe company’s short-term objectives are to:- continue to fund research into all types of eye diseases annually in Australia- continue to be in the forefront of advancing eye research in Australia - continue to support the presentation of research and the publication of the results of research for vision scientists and ophthalmologists for the benefit of all Australians.- continue to support new scientists by providing a percentage of its annual funding to support this category

The company’s long-term objectives are to:- increase the funds available for the provision of research funding in order to achieve its mission statement of advancing eye research in Australia. - ensure that the funding it provides leads to researchers gaining a track record to enable them to secure larger grants towards bigger and successful projects.

FINANCIALS


7. STRATEGIESTo achieve its stated objectives, the company has adopted the following strategies:

- The company is partnered with the Eye Sugeons’ Foundation who are now primarily responsible for raising additional funding towards the ORIA’s research projects and to raise awareness generally of eye health within Australia.

- The company’s Investment Advisory Committee monitors and works towards successfully managing the company’s invested funds, the profits from which are used annually for research funding.

- The company connects with other vision related organisations in Australia to support funding of projects for specific diseases.

- The company strives to attract, support and retain quality staff who are committed to the work of the organisation.

- The company conducts audits on its previously funded research to ensure the funding it provides is meeting its objectives.

- The company’s Board and Research Advisory Committee is made up of leading vision scientists and ophthalmologists within Australia.

8. OPERATING RESULTS(1) OPERATING REVENUERevenue is mainly derived from investing in shares and interest bearing securities.

2017 2016

Net dividend, interest and trust distribution income

$ 690,524 $ 559,635

Less Expenses 47,156 40,513

$ 643,368 $ 519,122

(2) OPERATING SURPLUSThe surplus of the company before other comprehensive income for the year ended 30 June 2017 was $1,989,919 (2016 $560,192). This amount is comprised of the following:

2017 2016

Trust Fund $ 2,011,277 $ 571,170

Administration (21,795) (10,978)

$ 1,989,482 $ 560,192

Other comprehensive income before grants and Director of Research allocation amounted to a profit of $568,764 (2016: deficit of $555,396) and included a loss on re arrangement of investments of $99,081 (2016: loss of $357,998) and valuation gain on available-for-sale financial assets of $667,845 (2016: loss of $197,398).

45Financials

9. REVIEW OF OPERATIONS The surplus for the year was $1,989,482 compared to $560,192 in 2016. Distributions from legacies and donations increased to $1,366,328 from $66,471 in 2016. This increase is due to two large bequests, $1,193,840 from The Anselmi Estate and $103,721 from The Estate of the late Mary H Tilden during the 2017 financial year. The administrative operations of the institute for the year resulted in a deficit of $21,795 compared with a deficit of $10,978 in 2016.

10. DIVIDENDSThe company’s Articles of Association preclude the payment of dividends to any of its members.

11. STATE OF AFFAIRSThere has been no significant change in the state of affairs of the company occurring during the year.

12. LIKELY DEVELOPMENTSAt the date of this report, there are no known unusual developments that will affect the results of the company’s operations in subsequent financial years.

13. SHARE OPTIONSNo share options were issued during the year.

14. DIRECTORS’ BENEFITSWith the exception of the grants made or allocated to Professor Stuart Graham, Clinical Professor Stephanie Watson, Proessor Peter McCluskey, Professor Jonathan Crowston, Dr Fred Chen and Professor Mark Gillies, no director of the company has since the end of the previous financial year, received or become entitled to receive a benefit not disclosed in the accounts as directors’ emoluments by reason of a contract made by the company or a related corporation with the directors, or with a firm in which he or she has a substantial financial interest.

15. AUDITOR’S INDEPENDENCE DECLARATIONA copy of the auditor’s independence declaration as required under the Australian Charities and Not-for-profits Commission Act 2012 is set out at page 27.

For and on behalf of the Board.

Clinical Professor Stephanie Watson Associate Professor Paul Healey Director Director

Sydney

Signed in accordance with a resolution of directors,

this 9th day of September 2017

FINANCIAL STATEMENTS


STATEMENT OF FINANCIAL POSITIONAS AT 30 JUNE 2017

Note2017

$2016

$

Current Assets

Cash and Cash Equivalents 3 2,554,053 1,323,579

Receivables 4 180,522 143,451

Investments 5 9,486,908 8,850,342

12,221,484 10,317,372

Non-Current Assets

Plant & Equipment 6 3,720 6,717

Total Assets 12,225,204 10,324,089

Current Liabilities

Payables 7 490,637 500,874

Provisions 8 23,310 27,999

513,947 528,873

Net Assets 11,711,257 9,795,216

Equity

General Fund 13 (a) - -

Capital Funds

Research Fund 9 2,438,183 1,236,057

Settled Funds 10 472,556 472,556

Financial Assets Reserve 11 1,580,380 912,535

Capitalised Profit on Re-arrangement of Investments, Capital Distributions & Transfers

12 6,695,085 6,794,166

11,186,204 9,415,314

Retained Income- Available for grants 13 (b) 525,053 379,902

Total Equity 11,711,257 9,795,216

The accompanying Notes form part of these financial statements.

47Financials

TRUST FUND STATEMENT OF COMPREHENSIVE INCOMEFOR THE YEAR ENDED 30 JUNE 2017

Note2017

$2016

$

Income

Dividends received from:

Other Corporations 608,087 491,290

Adjustment for 2015 imputation credits (1,991) (48,138)

Total Dividends 606,096 443,152

Interest received from:

Other Entities 21,236 48,527

Trust distributions received from:

Other Entities 63,192 67,956

The Richard and Ina Humbly Foundation 65,502 - 756,026 559,635

Legacies - Anselmi Estate 1,193,840 53,197

- Ivy May Stephenson 3,265 3,758

- The Estate of the late Gladys Clare Dickson Bequest - 9,516

- The Estate of The Late Mary H Tilden 103,721 -

Sundry Income 1,581 678

Total Income for the Year 2,058,433 626,784

Expenses

Legal Fees - 15,101

Commission Paid 47,156 40,513

47,156 55,614

Surplus For The Year 2,011,277 571,170

Other Comprehensive Income

Valuation Gains/(Losses) on available-for-sale financial assets 667,845 (197,398)

Profit/(Loss) on Re-arrangement of Investments (99,081) (357,998)

Total other comprehensive income 568,764 (555,396)

Surplus for the year before allocation 2,580,041 15,774

Grants Allocated/made during the year 14 439,205 414,750

Allocation to Director of Research - Victoria 15 203,000 157,000

642,205 571,750

Total Comprehensive Income/(Loss) 1,937,836 (555,976)

Profit/(Loss) Attributable to Members of the Entity 1,369,072 (580)

Total Other Comprehensive Income/(Loss) Attributable to Members of the Entity 568,764 (555,396)




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49Financials

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ADMINISTRATION STATEMENT OF COMPREHENSIVE INCOMEFOR THE YEAR ENDED 30 JUNE 2017

Note2017

$2016

$

Income

Membership Fees - RANZCO 150,650 147,696

Total Income 150,650 147,696

Expenses

Accountancy Fees 31,451 23,354

Auditors' Remuneration 16 5,750 4,950

Bank Charges 190 207

Consulting Fees 10,000 -

Depreciation 2,997 1,942

General Expenses 9,802 7,931

IT & Webpage Expenses 1,912 493

Insurance 2,164 2,798

Printing & Stationery 5,771 4,599

Loss on Disposal of Equipment - 14

Staff Salaries 52,042 47,208

Superannuation Contribution 9,269 9,025

Salary Sacrificed Benefits 25,200 28,000

Provision Employee Benefits (4,689) 6,434

Meeting and Travelling Expenses 20,586 21,719

Total Expenses 172,445 158,674

Surplus/(Deficit) For The Year 13 (a) (10,978) (268)

Other Comprehensive Income - -

Total Comprehensive Income (21,795) (10,978)




STATEMENT OF CASH FLOWSFOR THE YEAR ENDED 30 JUNE 2017

Note2017

$2016

$

Cash Flows From Operating Activities

Receipts

Dividends Received 570,258 489,093

Interest Received 21,236 48,527

Trust Distributions 127,461 67,956

Legacies 1,300,826 66,471

Other Revenue 3,572 957

RANZCO - Reimbursement of membership fees 150,650 142,150

Contributions from Eye Surgeons Foundation (Formerly RANZCO Eye Foundation)

100,000 100,000

Contributions from RANZCO NSW Branch - 24,750

Payments

Commissions (47,156) (40,513)

Research Grants Paid (591,352) (694,249)

Payments to Director of Research - Victoria (157,000) (210,000)

Other (180,221) (169,646)

Net Cash (Used in)/Provided by Operating Activities 17 1,298,275 (168,958)

Cash Flows From Investing Activities

Proceeds from Re-arrangement of Investments 551,493 1,285,941

Payments for Property, Plant & Equipment - (6,399)

Payments for Investments (619,294) (1,607,995)

Net Cash Used in Investing Activities (67,801) (328,453)

Net(Decrease)/Increase in Cash and Cash Equivalents 1,230,474 (497,411)

Cash and Cash Equivalents at 1 July 2016 1,323,579 1,820,990

Cash and cash equivalents at 30 June 2017 3 2,554,053 1,323,579


51Financials

NOTES TO AND FORMING PART OF THE FINANCIAL STATEMENTS

1 STATEMENT OF ACCOUNTING POLICIESThe financial statements are for the Ophthalmic Research Institute of Australia, incorporated and domiciled in Australia. The Ophthalmic Research Institute of Australia is a company limited by guarantee.

(A) BASIS OF PREPARATIONThe financial statements are general purpose financial statements that have been prepared in accordance with Australian Accounting Standards (including Australian Accounting Interpretations) and the Australian Charities and Not-for-profits Commission Act 2012.

The accounting policies set out below have been consistently applied to all years presented, unless otherwise stated. The financial report has been prepared on an accruals basis and is based on historical costs and does not take into account changing money values or, except where stated, current valuations of non current assets. Cost is based on the fair values of the consideration given in exchange for assets.

The following is a summary of the significant accounting policies adopted by the company in the preparation of the financial report.

(B) INCOME TAX

The company is an approved research institute and is exempt from income tax.

(C) TRANSFERS TO CAPITAL FUNDS

(i) Capital profits and losses on disposal of investments & capital distributions.

Realised capital profits and losses on disposal of investments are brought to account in the trust fund as profit/(loss) on rearrangement of investments, however, these amounts are transferred to capital funds and do not form part of retained income available for grants.

Capital Distributions and special dividends together with associated imputation credits recognised in the statement of comprehensive income are also transferred to the capital fund and do not form part of retained income available for grants.

(ii) General Research Capital FundTen percent of the net surplus of the General Fund including imputation credits are transferred to the General Research Capital Fund this financial year.

(iii) Allocation of Income to Each Fund

During the year ended 30 June 1993, the investments of the company were separated into the D.W. Research Fund and the General Fund in the ratio of 72% and 28% respectively. As the flow of investment and donation income to and from the two funds does not occur in the same proportion, the ratio of the D.W. Research Fund and the General Fund is no longer at 72% and 28%.

Income from the General Fund which comprises of all funds except the D.W. Research Fund, is allocated as follows:

Research Fund 10.0%

Esme Anderson 51.4%

G.J.Williams 8.9%

B. Mitchell 8.9%

Dame Ida Mann 12.5%

R. & L. Lowe Research 8.3%



If and when further donations are received by specific fund(s) the allocation of future income will be distributed to each fund in accordance with its revised proportion to the General Fund.

Fifty percent of the income derived from the D.W. Research Fund and its investments is allocated to the Director of Research Victoria.

(D) CASH AND CASH EQUIVALENTS

For the purpose of the statement of cash flows, cash and cash equivalents include cash on hand and at call deposits with banks.

(E) INVESTMENTS

Investments are carried at fair value. Changes in fair value will be held in an equity reserve until the asset is disposed, at which time the changes in fair value will be brought to account through the statement of comprehensive income.

(F) REVENUE

Interest and dividends are recognised when received.Grants, donations and distributions income are recognised when received.

(G) GOODS AND SERVICES TAX (GST)

All revenue, expenses and assets are recognised net of the amount of goods and services tax (GST), except where the amount of GST incurred is not recoverable from the Australian Tax Office. In these circumstances the GST is recognised as part of the cost of acquisition of the asset or as part of an item of the expense. Receivables and payables in the statement of financial position are shown inclusive of GST.

(H) FINANCIAL INSTRUMENTS

Recognition and Initial Measurement

Financial instruments, incorporating financial assets and financial liabilities, are recognised when the entity becomes a party to the contractual provisions of the instrument.

Financial instruments are initially measured at fair value plus transactions costs where the instrument is not classified as at fair value through profit or loss. Financial instruments are classified and measured as set out below.

Classification and Subsequent Measurement

(i) Loans and receivablesLoans and receivables are non-derivative financial assets with fixed or determinable payments that are not quoted in an active market and are subsequently measured at amortised cost using the effective interest rate method.

(ii) Held-to-maturity investmentsHeld-to-maturity investments are non-derivative financial assets that have fixed maturities and fixed or determinable payments, and it is the entity’s intention to hold these investments to maturity. They are subsequently measured at amortised cost using the effective interest rate method.

(iii) Available-for-sale financial assetsAvailable-for-sale financial assets are non-derivative financial assets that are either designated as such or that are not classified in any of the other categories. They comprise investments in the equity of other entities where there is neither a fixed maturity nor fixed or determinable payments.

(iv) Financial liabilitiesNon-derivative financial liabilities (excluding financial guarantees) are subsequently measured at amortised cost using the effective interest rate method.

53Financials

Fair value

Fair value is determined based on current bid prices for all quoted investments. Valuation techniques are applied to determine the fair value for all unlisted securities, including recent arm’s length transactions, reference to similar instruments and option pricing models.

Impairment

At each reporting date, the entity assesses whether there is objective evidence that a financial instrument has been impaired. In the case of available-for-sale financial instruments, a prolonged decline in the value of the instrument is considered to determine whether an impairment has arisen. Impairment losses are recognised in the statement of comprehensive income.

(I) IMPAIRMENT OF ASSETS

At each reporting date, the entity reviews the carrying values of its assets to determine whether there is any indication that those assets have been impaired. If such an indication exists, the recoverable amount of the asset, being the higher of the asset’s fair value less costs to sell and value in use, is compared to the asset’s carrying value. Any excess of the asset’s carrying value over its recoverable amount is expensed to the statement of comprehensive income.

Where it is not possible to estimate the recoverable amount of an individual asset, the entity estimates the recoverable amount of the cash-generating unit to which the asset belongs.

2 MEMBERS’ GUARANTEEIf the company is wound up the Memorandum of Association states that each member is required to contribute a maximum of $ 2.00 each towards meeting any outstanding obligations of the company.

CASH AND CASH EQUIVALENTS

2016 $

2015 $

General Account 2,046,907 985,929

Donations Account - 67,920

D.W. Research Fund Account 507,146 269,730

2,554,053 1,323,579

4 RECEIVABLES

Sundry Debtors 180,522 143,451

180,522 143,451

5 INVESTMENTS

Shares in Listed Corporations & Other Securities 9,486,908 8,850,342

Total Available-for-sale Financial Assets 9,486,908 8,850,342

Total Investments 9,486,908 8,850,342



6 PLANT AND EQUIPMENT

Office Equipment - at cost 13,151 13,151

Less: Accumulated Depreciation (9,431) (6,434)

3,720 6,717

Reconciliation

Reconciliation of the carrying amount of plant and equipment at the beginning and end of the current and previous financial year:

Carrying amount at beginning of year 6,717 2,274

Additions - 6,399

Disposal of Equipment - (14)

Less: Depreciation expense (2,997) (1,942)

Carrying amount at end of year 3,720 6,717

7 PAYABLES

Creditors and Accruals 18,033 22,123

Grants Payable 269,604 321,751

Director of Research - Victoria (refer note 15) 203,000 157,000

490,637 500,874

8 PROVISIONS

Employee Benefits 23,310 27,999

9 RESEARCH CAPITAL FUND

General

Balance 1 July 2016 914,702 939,933

Allocation to Capital:

- 10% Surplus & Imputation Credits & Other Legacies 42,507 24,769

- Capitalised Bequests 1,159,619 -

Transfer from Capital:

-Amount transferred to Income - (50,000)

Balance 30 June 2017 2,116,828 914,702

Anselmi Estate

Balance 1 July 2016 290,979 290,979

Allocation during year - -

Transfer during year - -

Balance 30 June 2017 290,979 290,979

2017 $

2016 $

55Financials

Ivy May Stephenson Estate

Balance 1 July 2016 30,376 30,376

Allocation during the year - -

Transfer during year - -

Balance 30 June 2017 30,376 30,376

Total 2,438,183 1,236,057

10 SETTLED FUNDS

D.W. Research Funds 200,000 200,000

Esme Anderson 124,326 124,326

G.J. Williams 25,500 25,500

B. Mitchell 26,023 26,023

Dame Ida Mann (Est. 31/03/84) 56,707 56,707

Ronald & Lois Lowe 40,000 40,000

472,556 472,556

11 FINANCIAL ASSETS RESERVE

Balance 1 July 2016 912,535 1,109,933

Revaluation increment/(decrement) 667,845 (197,398)

Balance 30 June 2017 1,580,380 912,535

Financial assets reserve records unrealised gains on revaluation of financial assets to fair value.

2017 $

2016 $



12 CAPITALISED PROFIT ON RE-ARRANGEMENT OF INVESTMENTS, CAPITAL DISTRIBUTIONS & TRANSFERS

Balance 30/06/16

$

Allocation of Realised Profit/(Loss) on

Rearrangement of Investments &

Capital Distributions & Transfers

$

Balance 30/06/17

$

Research Fund

General 129,761 (2,975) 126,786

Anselmi Estate 46,478 (1,065) 45,413

Ivy May Stephenson 120 (3) 117

D.W. Research Funds 5,034,238 (58,653) 4,975,585

Esme Anderson 914,479 (20,780) 893,699

G.J. Williams 157,054 (3,598) 153,456

B. Mitchell 155,118 (3,598) 151,520

Dame Ida Mann 219,106 (5,053) 214,053

Ronald & Lois Lowe 137,812 (3,356) 134,456

6,794,166 (99,081) 6,695,085

13 ACCUMULATED FUNDS

(a) Administration

Accumluated Deficits - 1 July 2016 - -

Total Comprehensive Income (21,795) (10,978)

Total available for appropriation (21,795) (10,978)

Aggregate of amounts transferred from Administration 13 (b) 21,795 10,978

Accumulated Deficits - 30 June 2017 - -

(b) Trust Fund

Retained income - 1 July 2016 379,902 416,229

Total Comprehensive Income 1,369,072 (580)

Total available for appropriation 1,748,974 415,649

Aggregate of amounts transferred to General/Capital Funds

Administration 13 (a) (21,795) (10,978)

Research Trust (1,202,126) (24,769)

Retained income - 30 June 2017 525,053 379,902

2017 $

2016 $

57Financials

14 GRANTS ALLOCATED / MADE DURING THE YEAR

Dr S McLenachan & *Dr F Chen - 50,000

Dr N Van Bergen, A/Prof I Trounce & Prof F Grus - 49,000

Prof J Smith - 49,000

Dr R Natoli, A/Prof R Essex & Prof J Provis - 48,000

Dr D M Bukowska & Dr E Wong - 50,000

Dr I L Sanchez - 45,000

Dr Jennifer Fan Gaskin - 50,000

Dr R Wong, Mr D Crombie, A/Prof A Pebay - 50,000

Dr A Cook & A/Prof Alex Hewitt - 50,000

Dr Z Wu & *Prof J Crowston - 49,000

Dr J Wood - 50,000

Dr S H Chung & *Prof M Gillies - 49,500

Dr Guei-Sheung Liu, A/Prof Alex Hewitt, A/Prof Bang Bui, Dr Anna King & Dr Amy Fan Li

50,000.00 -

*Dr Fred Chen 50,000.00 -

Dr Sandy Hung & Mrs Sandra Staffieri 50,000.00 -

Dr Vivik Gupta & *Prof Stuart Graham 49,700.00 -

Prof Justine Smith 49,850.00 -

A/Prof R Max Conway, Dr Svetlana Cherepanoff, Dr Michael Giblin, Prof Richard Epstein, A/Prof Anthony Joshua, Dr Wenchan Wong & Ms Amparo Herrera-Bond

50,000.00 -

Dr Michele C Madigan & *Prof Peter J McCluskey 46,600.00 -

Dr Srujana Sahebjada 49,105.00 -

Dr Jia Jia Lek 47,650.00 -

Dr Yuyi You & A/Prof Alexander Klistorner 48,450.00 -

Dr Michelle Sun, A/Prof Andrea O’Connor & Dr John Wood 47,850.00 -

539,205 589,500

Deduct Contribution from:

The Ophthalmic Research Institute of Australia - 50,000

Eye Surgeons Foundation (Formerly RANZCO Eye Foundation) 100,000 100,000

RANZCO NSW Branch - 24,750

RANZCO - -

100,000 174,750

439,205 414,750

* Grant received by director

2017 $

2016 $



15 FUNDS ALLOCATED TO DIRECTOR OF OPHTHALMIC RESEARCH - VICTORIA

Balance as at 1 July 2016 157,000 210,000

Interest for the year 387 1,105

Allocation for year 203,000 157,000

360,387 368,105

Payment made to Director of Research 157,387 211,105

Balance as at 30 June 2017 203,000 157,000

16 AUDITORS REMUNERATION

Financial Statements - Audit Service 5,750 4,950

Other services - -

5,750 4,950

17 RECONCILIATION OF NET CASH PROVIDED BY OPERATING ACTIVITIES TO RESULTS FOR YEAR

Net Surplus/(Deficit)

- Trust Fund 1,937,836 (555,976)

- Administration (21,795) (10,978)

1,916,041 (566,954)

Depreciation 2,997 1,942

Disposal of Equipment - 14

Provision for Employee Benefits (4,689) 6,434

Transfer from Capital to Contribute Towards Grants - (50,000)

(Increase)/Decrease in Receivables (37,072) 46,219

Increase/(Decrease) in Creditors and Accrued Expenses (4,091) (4,260)

Increase/(Decrease) in Grants Payable (52,147) (104,749)

Increase/(Decrease) in allocation to Director of Research - Victoria 46,000 (53,000)

Valuation (Gains)/Losses on available-for-sale financial assets (667,845) 197,398

(Profit)/Loss on Rearrangement of Investments 99,081 357,998

Net Cash Provided by /(used in) Operating Activities 1,298,275 (168,958)

2017 $

2016 $

59Financials

18 DISCLOSURES ON DIRECTORS AND OTHER KEY MANAGEMENT PERSONNELDIRECTORSThe following directors received grants during the year. These are detailed at note 14.

Professor Stuart Graham

Clinical Professor Stephanie Watson

Professor Peter McCluskey

Professor Jonathan Crowston

Dr Fred Chen

Professor Mark Gillies

The names of the directors who have held office during the financial year are:Clinical Professor Stephanie Watson, Sydney (Chairman)

Professor Mark Gillies, Sydney (Vice Chairman)

Professor Richard Mills, Adelaide (Honorary Secretary)

Associate Professor Paul Healey, Sydney (Honorary Treasurer)

Dr Wislon Heriot, Melbourne

Dr Fred Chen, Perth

Professor Jonathon Crowston, Melbourne



Professor Stuart Graham, Sydney


Professor David Mackey, Perth

Professor Paul Mitchell, Sydney

Professor Peter J McCluskey, Sydney


Associate Professor Andrea Vincent, New Zealand

KEY MANAGEMENT PERSONNELOther Key Management Personnel include Executive Officer, Anne Dunn Snape.

Key management personnel are those persons having authority and responsibility for planning, directing and controlling the activities of the entity, directly or indirectly, including any director (whether executive or otherwise) of that entity. Control is the power to govern the financial and operating policies of an entity so as to obtain benefits from its activities.

KEY MANAGEMENT PERSONNEL COMPENSATIONKey Management Personnel has been taken to comprise the directors and one member of the executive management responsible for the day to day financial and operational management of the entity.

2017 $

2016 $

(a) Short-term employee benefits 77,242 75,208

(b) Post-employment benefits 9,269 9,025

(c) Other long-term benefits - -

(d) Termination benefits - -

(e) Share-based payment - -

86,511 84,233

2017 $

2016 $



19 FINANCIAL INSTRUMENTS

(A) FINANCIAL RISK MANAGEMENT POLICIESThe entity’s financial instruments consist mainly of deposits with banks, local money market instruments, short-term investments, accounts receivable and payable.

The entity does not have any derivative instruments at 30 June 2017.

(i) Treasury Risk Management An investment committee consisting of Board members of the entity meet on a regular basis to analyse

financial risk exposure and to evaluate treasury management strategies in the context of the most recent economic conditions and forecasts.

The committee’s overall risk management strategy seeks to assist the entity in meeting its financial targets, whilst minimising potential adverse effects on financial performance.

Risk management policies are approved and reviewed by the Board on a regular basis. These include credit risk policies and future cash flow requirements.

(ii) Financial Exposures and Management Risk The main risks the entity is exposed to through its financial instruments are interest rate risk, liquidity risk and

credit risk.

Interest rate risk Interest rate risk is managed with a mixture of fixed and floating rates on investments.

Foreign currency risk The entity is not exposed to fluctuations in foreign currencies.

Liquidity risk The entity manages liquidity risk by monitoring forecast cash flows.

Credit risk The maximum exposure to credit risk, excluding the value of any collateral or other security, at balance date to recognised financial assets, is the carrying amount, net of any provisions for impairment of those assets, as disclosed in the statement of financial position and notes to the financial statements.

The entity does not have any material credit risk exposure to any single receivable or group of receivables under financial instruments entered into by the entity.

Price risk The group is not exposed to any material commodity price risk.

61Financials

(B) F

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2016

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2017

%

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List

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Bills

N/A

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--

--

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180,

522

143,

451

180,

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554,

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9,66

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3,79

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2,55

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31,

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--

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9,17

9,23

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8,49

2,91

9

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,283

9,

816,

498



(C) NET FAIR VALUESThe net fair values of listed investments have been valued at the quoted market bid price at balance date. For other assets and other liabilities the net fair value approximates their carrying value. No financial assets and financial liabilities are readily traded on organised markets in standardised form other than listed investments.

The aggregate net fair values and carrying amounts of financial assets and financial liabilities are disclosed in the statement of financial position and in the notes to and forming part of the financial statements.

(D) SENSITIVITY ANALYSIS

Interest Rate RiskThe entity has performed a sensitivity analysis relating to its exposure to interest rate risk at balance date. This sensitivity analysis demonstrates the effect on the current year results and equity which could result from a change in this risk.

Interest Rate Sensitivity Analysis:At 30 June 2017, the effect on profit and equity as a result of changes in the interest rate, with all other variables remaining constant, would be as follows:

2017Carrying Amount

Interest Rate Risk

$ -1% Profit +1% Profit -1% Equity +1% Equity

Financial Assets

Cash and Cash Equivalents 2,554,053 (25,541) 25,541 (25,541) 25,541

2016Carrying Amount

Interest Rate Risk

$ -1% Profit +1% Profit -1% Equity +1% Equity

Financial Assets

Cash and Cash Equivalents 1,323,579 (13,236) 13,236 (13,236) 13,236

63Financials

20 FAIR VALUE MEASUREMENTS

Financial assets and financial liabilities measured at fair value in the statement of financial position are grouped into three Levels of a fair value hierarchy. The three Levels are defined based on the observability of significant inputs to the measurement, as follows:

Level 1: quoted prices (unadjusted) in active markets for identical assets or liabilities;

Level 2: inputs other than quoted prices included within Level 1 that are observable for the asset or liability, either directly or indirectly;

Level 3: unobservable inputs for the asset or liability.

The following table shows the Levels within the hierarchy of financial assets and liabilities measured at fair value on a recurring basis at 30 June 2017 and 30 June 2016:

NoteLevel 1

$Level 2

$Level 3

$Total

$

30 June 2017

Assets

Listed securities 5 9,486,908 - - 9,486,908

Net fair value 9,486,908 - - 9,486,908

30 June 2016

Assets

Listed securities 5 8,850,342 - - 8,850,342

Net fair value 8,850,342 - - 8,850,342

There were no transfers between Level 1 and Level 2 for assets measured at fair value during 2017 or 2016.

Listed SecuritiesFair values have been determined by reference to their quoted bid prices at the reporting date.

21 BENEFICIARY ENTITLEMENT

The company is a beneficiary of The Richard & Ina Humbley Foundation and has an entitlement to income from the foundation to be used for grants in support of research conducted into macular degeneration.

The company accounts for this income on a cash basis.

The income received for the year ended 30 June 2017 from the foundation was $65,502.



DIRECTORS’ DECLARATION

The directors of the company declare that:

1. the financial statements and notes as set out on pages 5 - 23:

(a) comply with Accounting Standards and Australian Charities and Not-for-profits Commission Act 2012; and

(b) give a true and fair view of the financial position as at 30 June 2015 and performance for the year ended on that date of the company.

2. In the directors’ opinion there are reasonable grounds to believe that the company will be able to pay its debts as and when they become due and payable.

The declaration is made in accordance with a resolution of the board of Directors.

On behalf of the Board.

Clinical Professor Stephanie Watson Associate Professor Paul Healey Director Director

Sydney, this 9th day of September 2017

65Financials

AUDITOR’S REPORT


AUDITOR’S REPORT

Editor: Anne Dunn SnapeDesign & Print Management: The Burrow Group

THE OPHTHALMICRESEARCH REPORTINSTITUTE OF AUSTRALIA


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