Biomarkers of canine glaucoma Kathleen L. Graham A thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy Clinical Ophthalmology and Eye Health Sydney Medical School The University of Sydney 2019 P a g e | 2 DECLARATION This thesis is submitted to the University of Sydney in fulfilment of the requirement for the degree of Doctor of Philosophy. This is to certify that the content of this thesis is my own work. This thesis has not previously been submitted for any degree or other purposes. I certify that the intellectual content of this thesis is the product of my own work and that all the assistance received in preparing this thesis and sources has been acknowledged. Kathleen Graham 27/09/2019 P a g e | 3 DISCLOSURE AND AUTHOR CONTRIBUTIONS The following chapters have been re-formatted from manuscripts published or under review in peer-reviewed journals. For each of the manuscripts I was responsible for study design; obtaining and complying with animal ethics approval and requirements; recruitment; conducting clinical examinations and imaging; collating and interpreting data; statistical analysis; manuscript composition and submission. Co-authors Associate Professor Andrew White, Dr Christina McCowan, Dr Eve Diefenbach, Dr Pip Johnson, Professor Jacky Reid, Professor Mitchell Lawlor, Erica Barry, Dr Cameron Whittaker, Dr Evelyn Hall, Dr Kelly Caruso, Sarah-Elizabeth Byosiere, Lynna Feng, Dr Matthew Saunders, Professor Pauleen Bennett, Usha Pattamatta, Marisa Perez Orrico, Domenic Soligo, Dr Mark Billson, Dr David Donaldson, Dr Charles Caraguel, and Professor Frank Billson assisted with conduct of the studies, analyses and revision of the manuscripts. The manuscripts produced for this thesis are as follows: CHAPTER 1 Graham KL, McCowan C, White A. Genetic and biochemical biomarkers in primary canine glaucoma. Veterinary Pathology 2016; 54 (2): 194-203 doi: 10.1177/030098581666661. CHAPTER 2 Graham KL, Reid J, Whittaker CJG, Hall EJS, Caruso K, McCowan CI, White A. Development of a vision impairment score for the assessment of functional vision in dogs: initial evidence of validity, reliability and responsiveness. Veterinary Ophthalmology. 30th January 2019, DOI: 10.1111/vop.12656. CHAPTER 3 Graham KL, Byosiere S-E, Feng LC, Sanders M, Bennett PC, Caruso K, McCowan CI, White A. A forced-choice preferential looking task for the assessment of vision in dogs: a pilot study. Journal of Small Animal Practice 2019; 60(6): 340-347 doi: 10.1111/jsap.12965 . CHAPTER 4 Graham KL, McCowan CI, White AJR. A modified protocol for assessment of the pupillary light reflex in dogs predisposed to glaucoma. Journal of Small Animal Practice. Under review CHAPTER 5 Graham KL, Whittaker CJG, Caruso K, McCowan CI, White A. Anterior segment ultrasound biomicroscopy of the unaffected eye in dogs with primary angle closure glaucoma. Veterinary Ophthalmology. Under review CHAPTER 6 Graham KL, McCowan CI, Caruso K, Billson FM, Whittaker CJG, White A. Optical coherence tomography of the retina, nerve fiber layer and optic nerve head in dogs with glaucoma. Veterinary Ophthalmology. 16th June 2019 DOI: 10.1111/vop.12694. CHAPTER 7 Graham KL, Johnson PJ, Barry E, Perez Orrico M, Soligo D, Lawlor M, White AJR. Diffusion tensor imaging of the visual pathway in dogs with primary angle closure glaucoma. Veterinary Ophthalmology. Under review P a g e | 4 CHAPTER 8 Graham KL, Diefenbach E, McCowan CI, White AJR. Shotgun proteomic analysis of the precorneal tear film in dogs with primary glaucoma. Veterinary Ophthalmology. Under review CHAPTER 9 Graham KL, Donaldson D, Billson FA, Billson FM. Use of a 350mm2 Baerveldt glaucoma drainage device to maintain vision and control intraocular pressure in dogs with glaucoma: a retrospective study (2013-2016). Veterinary Ophthalmology. 2017; 20(5): 427-434. DOI: 10.1111/vop.12443. CHAPTER 10 Graham KL, Hall EJS, Caraguel C, White A, Billson FA, Billson FM. Comparison of diode laser trans-scleral cyclophotocoagulation versus implantation of a 350mm2 Baerveldt glaucoma drainage device for the treatment of glaucoma in dogs (a retrospective study: 2010-2016). Veterinary Ophthalmology. 2018; 21(5): 487-497 DOI: 10.1111/vop.12536. FORMAL PRESENTATIONS Diffusion tensor imaging of the visual pathway in dogs with angle closure glaucoma. American College of Veterinary Radiology annual conference; Baltimore Maryland, USA. November 2019 Posterior segment optical coherence tomography in dogs predisposed to primary glaucoma. American College of Veterinary Ophthalmology annual conference; Minneapolis Minnesota, USA. September 2018 Biomarkers in canine glaucoma – structure and function. Australian and New Zealand College of Veterinary Scientists Annual Conference; Ophthalmology chapter; Gold Coast Queensland. July 2018 Optics of vision. Royal Veterinary College seminar; Royal Veterinary College, University of London; Hertfordshire, United Kingdom. October 2017 Vision assessment in dogs. UNSW School of Psychology Vision Research presentation; University of New South Wales, Randwick NSW. May 2017 Biomarkers of canine glaucoma. Save Sight Institute Annual Research Symposium; Sydney Eye Hospital Sydney NSW. January 2017 P a g e | 5 DECLARATION BY SUPERVISOR This is to certify that all co-authors have provided their consent for inclusion of the manuscripts presented in this thesis. The co-authors accept the Kathleen’s contribution to each manuscript and the description of the co-authors’ contribution. Clinical Associate Professor Andrew White 30/09/2019 P a g e | 6 ACKNOWLEDGEMENTS There are many people who have been instrumental in the development and conduct of this project. Most importantly, I want to acknowledge the dogs that were included and their owners who saw the value of participating in this clinical research. To my supervisor, Clin. A/Professor Andrew White, I am grateful for your voice of reason, practical approach to clinical research, and your eternal optimism. To my auxiliary supervisor Dr Christina McCowan, I am grateful for your common sense, grammatical corrections, and for your interest in the details and all the possibilities. To all the veterinarians and nurses who allowed me to spend time in their hospitals, meet their clients, and examine their patients, especially Dr Anne Fawcett, Dr Belinda Parsons and Dr Kelly Caruso. I very much appreciate being allowed into your hospitals to work with your clients and your patients. And to those who collaborated on the studies and made them a reality, I remain grateful. Especially Dr Philippa Johnson from Cornell University College of Veterinary Medicine for sharing her talents and kindness; Dr Eve Diefenbach from the Westmead Institute for Medical Research proteomics department for her patience, time, knowledge, skill and for always seeing the funny side; and Professor Jacky Reid for her time and expertise. P a g e | 7 TABLE OF CONTENTS DECLARATION ................................................................................................................. 2 DISCLOSURE AND AUTHOR CONTRIBUTIONS .................................................................... 3 FORMAL PRESENTATIONS ................................................................................................ 4 DECLARATION BY SUPERVISOR ........................................................................................ 5 ACKNOWLEDGEMENTS .................................................................................................... 6 TABLE OF CONTENTS ....................................................................................................... 7 ABSTRACT ..................................................................................................................... 13 LIST OF FIGURES ............................................................................................................ 15 LIST OF TABLES .............................................................................................................. 23 LIST OF ABBREVIATIONS ................................................................................................ 26 CHAPTER ONE AIM OF THE THESIS AND LITERATURE REVIEW Literature review ............................................................................................................ 32 What is glaucoma? ...................................................................................................... 32 Classification ............................................................................................................... 33 Aetiology ..................................................................................................................... 33 Iridocorneal angle morphology ................................................................................... 38 Stage of disease ........................................................................................................... 39 Aqueous humour dynamics ........................................................................................ 39 Genetic and biochemical biomarkers in primary canine glaucoma ............................... 43 Abstract ....................................................................................................................... 43 Introduction ................................................................................................................ 43 Genetic markers .......................................................................................................... 45 Biochemical markers