2025 Annual Meeting and Alumni Reunion Program

2025 Annual Meeting and Alumni Reunion

June 12-13, 2025 I The Starr Center, Boston, MA

MEETING CO-CHAIRS

Joan W. Miller, MD

David Glendenning Cogan Professor of Ophthalmology and Chair, Department of Ophthalmology, Harvard Medical School; Chair of Ophthalmology, Mass Eye and Ear and Mass General Hospital; Ophthalmologist-in-Chief, Brigham and Women’s Hospital

Joseph B. Ciolino, MD

Associate Professor of Ophthalmology, Harvard Medical School; Associate Director of Harvard Ophthalmology Alumni; Henry Freeman Allen Cornea Scholar, Mass Eye and Ear

Eric Gaier, MD, PhD

Assistant Professor of Ophthalmology, Harvard Medical School; Site Director of the Harvard Ophthalmology Residency Program, Boston Children’s Hospital

Rachel Huckfeldt, MD, PhD

Assistant Professor of Ophthalmology, Harvard Medical School; Director, Inherited Retinal Degenerations Fellowship, Mass Eye and Ear

Joseph F. Rizzo III, MD

Simmons Lessell Professor of Ophthalmology, Harvard Medical School; Director of Harvard Ophthalmology Alumni, Co-Director, Harvard Ophthalmology Mobility Enhancement and Vision Rehabilitation Center of Excellence; Director, Neuro-Ophthalmology Service, Mass Eye and Ear

Annual Meeting and Alumni Reunion

ANNUAL MEETING

Thursday, June 12, 2025 - The Starr Center - Second Floor, 185 Cambridge Street, Boston, MA

12:00-12:25 pm

12:25–12:30 pm

12:30–12:35 pm

12:35–1:05 pm

Registration & Lunch

Welcome & Annual Meeting Overview

Rachel Huckfeldt, MD, PhD (2013, 2016) and Eric D. Gaier, MD, PhD (2017, 2019) - Meeting Co-Chairs

Introduction of the 2025 Alumni Grand Rounds Speaker

Eric D. Gaier, MD, PhD (2017, 2019) - Assistant Professor of Ophthalmology, Harvard Medical School; Site Director of the Harvard Ophthalmology Residency Program, Boston Children’s Hospital

1:05–1:17 pm

1:17–1:29 pm

1:29–1:41 pm

1:41–1:53 pm

1:53–2:05 pm

2:05–2:15 pm

2:15–2:20 pm

2025 Alumni Grand Rounds Lecture

Innovations in Retina Surgery Visualization and Imaging

Katherine E. Talcott, MD (2015) - Associate Professor of Ophthalmology, Lerner College of Medicine; Associate Residency Program Director and Retinal Surgeon, Cole Eye Institute, Cleveland Clinic

Surgical Advances in Pediatric Ptosis Surgery: The Frontalis Flap

Edith Reshef, MD (2020, 2022) - Instructor in Ophthalmology, Harvard Medical School and Ophthalmologist, Boston Children's Hospital

Grieving an Ease of Living: Psychosocial Perspectives on the Impact of Vision Loss

Ethan G. Lester, PhD - Assistant Professor of Psychology and Assistant Professor of Ophthalmology, Harvard Medical School and Clinical Psychologist, Massachusetts General Hospital

Antibiotic-Resistant Infection Can Complicate Even the Finest Surgeries – New Advances

Michael S. Gilmore, PhD - Sir William Osler Professor of Ophthalmology, Harvard Medical School; Director, Infectious Harvard Ophthalmology Disease Institute; and Chief Scientific Officer, Mass Eye and Ear

Using Genetics in the Glaucoma Clinic

Janey L. Wiggs, MD, PhD (1990, 1991, 1992) - Paul Austin Chandler Professor of Ophthalmology, Harvard Medical School; Co-Director, Harvard Ophthalmology Glaucoma Center of Excellence; Associate Director, Howe Laboratory, and Associate Chief, Ophthalmology Clinical Research, Mass Eye and Ear

Impact of Concussion on Visual Function

Aparna Raghuram, OD, PhD - Assistant Professor of Ophthalmology, Harvard Medical School; Optometrist, Boston Children's Hospital

Harvard Ophthalmology Research Scholars Program and Introduction of the Class of 2025

Silas L. Wang, MD - Instructor in Ophthalmology, Harvard Medical School; Co-Director, Harvard Ophthalmology Research Scholars Program; and Physician and Surgeon, Mass Eye and Ear

The Harvard Ophthalmology Research Scholar Experience

Cherrell Price, MS (2021) - Medical School Candidate, Morehouse School of Medicine

Break 2:20-3:00 pm

3:00–3:12 pm

3:12–3:24 pm

Modern Approach for Diagnosis and Treatment of Retinoblastoma

Efren Gonzalez, MD (2014) - Assistant Professor of Ophthalmology, Harvard Medical School; Director, Ocular Oncology Center and Pediatric Vitreoretinal Surgeon, Boston Children's Hospital

Emerging Therapies for Inherited Retinal Disorders: How Do We Measure Success?

Rachel Huckfeldt, MD, PhD (2013, 2016) - Assistant Professor of Ophthalmology, Harvard Medical School; Director, Inherited Retinal Degenerations Fellowship, Mass Eye and Ear

3:24–3:36 pm

3:36–3:48 pm

3:48–4:00 pm

4:00–4:05 pm

4:05–4:15 pm

The Mass Eye and Ear Data and Imaging Repository and its Applications

Tobias Elze, PhD (2013) - Associate Professor of Ophthalmology, Harvard Medical School; Associate Scientist, Schepens Eye Research Institute of Mass Eye and Ear

What's New in Dry Eye?

Jennifer Luiz Lindsey, MD, MBA - Member of the Faculty of Ophthalmology, Harvard Medical School; Director, Harvard Ophthalmology Residency Training Program, Harvard Ophthalmology Vice Chair of Education, and Associate Chief for Ophthalmology Education and Diane Kaneb Chair in Ophthalmology, Mass Eye and Ear

Gene Therapy to Rescue Optic Neuropathy in Familial Dysautonomia

Anil K. Chekuri, PhD (2021) - Instructor in Ophthalmology, Harvard Medical School; Investigator, Mass Eye and Ear

Harvard Retinal Imaging Laboratory Undergraduate Mentoring Program Update

Edward S. Lu, MD (2026) - Ophthalmology Resident (PGY-3), Harvard Medical School; Co-founder, Harvard Retinal Imaging Laboratory Undergraduate Mentoring Program, Mass Eye and Ear

Perspective on Mariana Dieste Mead & Introduction of the 2025 Mariana Dieste Mead Lecturer

Joseph F. Rizzo III, MD (1986) - Simmons Lessell Professor of Ophthalmology, Harvard Medical School; Co-Director, Harvard Ophthalmology Mobility Enhancement and Vision Rehabilitation Center of Excellence; Director, Neuro-Ophthalmology Service, Mass Eye and Ear

4:15–4:50 pm

4:50–4:55 pm

2025 Mariana Dieste Mead Lecture

My Career in Ophthalmology, Adapting to Change

Jan A. Kylstra, MD (1987, 1989) - Former Lecturer on Ophthalmology, Harvard Medical School

Annual Meeting Closing Remarks

Rachel Huckfeldt, MD, PhD (2013, 2016) and Eric D. Gaier, MD, PhD (2017, 2019) - Meeting Co-Chairs

5:30-8:00 pm

Joan W. Miller, MD Celebration Reception

ACCREDITATION STATEMENT

In support of improving patient care, Boston Children’s Hospital is jointly accredited by the Accreditation Council for Continuing Medical Education (ACCME), the Accreditation Council for Pharmacy Education (ACPE), and the American Nurses Credentialing Center (ANCC), to provide continuing education for the healthcare team. Fees associated with AMA PRA Category 1 Credits™ have been provided by the HMS Department of Ophthalmology. Fees associated with COPE credit hours have been provided by the HMS Department of Ophthalmology. For complete CME information, visit: www.eventleaf.com/e/AMAR2025

OPHTHALMOLOGISTS: Physician Boston Children’s Hospital designates this live activity for a maximum of 12.0 AMA PRA Category 1 Credits™. Physicians should claim only credit commensurate with the extent of their participation in this activity.

OPTOMETRISTS: Optometric CE Boston Children’s Hospital designates this live activity for a maximum of 12.0 COPE Credit Hours. Optometrists should claim only the credit commensurate with the extent of their participation in the activity. The course number is 250612-JA

TEXT-IN-ATTENDANCE: Send PO76T to 844-998-2674 during the session to record your attendance.

The Langham Hotel - 250 Franklin Street, Boston, MA

2025 Annual Meeting and Alumni Reunion

ALUMNI REUNION

Friday, June 13, 2025 - The Starr Center - Second Floor, 185 Cambridge Street, Boston, MA

8:15-8:45 am

8:45–8:50 am

8:50 am

8:51–9:06 am

9:06

9:07–9:22 am

Registration & Breakfast

Welcome & Alumni Reunion Overview

Joseph F. Rizzo III, MD (1986) and Joseph B. Ciolino, MD (2009) - Meeting Co-Chairs

9:23–9:38 am

Ocular Surface Stem Cell Transplantation at Mass Eye and Ear

Thomas Dohlman, MD (2020) - Assistant Professor of Ophthalmology, Harvard Medical School; Medical Director, Boston Keratoprosthesis Program, Mass Eye and Ear

9:39–

Functional Imaging of the Extraocular Muscles

Nailyn Rasool, MD (2015) - Associate Professor of Ophthalmology, Harvey A. Birsner Endowed Chair in NeuroOphthalmology, and Director, Neuro-ophthalmology Fellowship, University of California, San Francisco

Education: An Investment with Infinite Return

Ankoor S. Shah, MD, PhD (2009, 2010, 2011) - Assistant Professor of Ophthalmology and Director of Ophthalmic Medical Student Education, Harvard Medical School; Director, Pediatric Ocular Trauma, Department of Ophthalmology, Boston Children's Hospital

Be the Solution in Healthcare: How You Should be Involved with Healthcare Leadership

Rishi P. Singh, MD (2005) - Professor of Ophthalmology, Lerner College of Medicine; Staff Surgeon, Cole Eye Institute; Vice President and Chief Medical Officer, Martin North and South Hospitals, Cleveland Clinic

10:41–11:01

11:02–11:22 am

11:22–11:27 am

25th Year: Life Post Mothership

Stella K. Kim, MD (2000) - Professor and Joe M. Green Jr. Chair in Ophthalmology, and Director, Cornea Department, Cizik Eye Clinic, University of Texas Health Science Center at Houston

11:27–11:59 am

How I Found My Professional Waze

Roberto Pineda II, MD (1994, 1995, 1996) - Associate Professor of Ophthalmology, Harvard Medical School; Thomas Y. and Clara W. Butler Chair in Ophthalmology, Mass Eye and Ear

Introduction of Distinguished Research Achievement Awardee

David M. Wu, MD, PhD - Assistant Professor of Ophthalmology, Harvard Medical School; Physician, Surgeon, and Joan Whitten Miller Scholar, Mass Eye and Ear

2025 Distinguished Research Achievement Award Lecture

Gene-agnostic Gene Therapy to Prolong Vision

Constance Cepko, PhD - Bullard Professor of Genetics and Neuroscience, Professor of Ophthalmology, and Principal Investigator of the Cepko Lab, Department of Genetics, and Co-Director of the Leder Human Biology and Translational Medicine Program; Harvard Medical School

1:05–1:25 pm

Department Update

Joan W. Miller, MD (1989, 1991) - David Glendenning Cogan Professor of Ophthalmology and Chair, Department of Ophthalmology, Harvard Medical School; Chair of Ophthalmology, Mass Eye and Ear and Mass General Hospital; Ophthalmologist-in-Chief, Brigham and Women’s Hospital

1:25 pm 1990 Class Speaker Introduction - James Harris, MD, PhD - PGY4 Resident, Harvard Medical School

1:26–1:46 pm

The Development of Botulin Toxins in Medicine: A Thousand Years of Scientific Discovery

Scott M. Whitcup, MD (1990) - Founder and CEO, Akrivista and Whitecap Biosciences

1:46 pm 1985 Class Speaker Introduction - Amee Azad, MD - PGY4 Resident, Harvard Medical School

1:47–2:07 pm

2:07–2:12 pm

2:12–2:45 pm

Lessons Learned in Medicine, Ophthalmology and Life: 40 Years Later

Daniel J. Townsend, MD (1984, 1985) - Instructor in Ophthalmology (Part-time), Harvard Medical School; Former President of the Medical Staff, Mass Eye and Ear

Introduction of Distinguished Clinical Achievement Awardee

Lotfi Merabet, OD, PhD, MPH - Associate Professor of Ophthalmology, Harvard Medical School; Co-director, Mobility Enhancement and Vision Rehabilitation Center of Excellence, Harvard Ophthalmology; Frederick and Thaddeus Jakobiec Chair in Ophthalmology and Harvard Ophthalmology Vice Chair for Basic and Translational Research, Mass Eye and Ear

2025 Distinguished Clinical Achievement Award Lecture

Therapeutic Neuro-Ophthalmology: From Oxymoron to Reality

Joseph F. Rizzo III, MD (1986) - Simmons Lessell Professor of Ophthalmology, Harvard Medical School; Director of Harvard Ophthalmology Alumni, Co-Director, Harvard Ophthalmology Mobility Enhancement and Vision Rehabilitation Center of Excellence; Director, Neuro-Ophthalmology Service, Mass Eye and Ear

3:30 pm 1980 Class Speaker Introduction - Ayush Parikh, MD - PGY2 Resident, Harvard Medical School

3:31–3:51 pm

3:51–4:16 pm

4:16–4:21 pm

4:21–4:55 pm

4:55–5:00 pm

The Newbury Hotel, Second Floor, 1 Newbury Street, Boston, MA Break 2:45-3:30 pm

Lifelong Learning

Dorothy Horns, MD (1980, 1981) - Adjunct Associate Professor, Department of Ophthalmology and Visual Neurosciences, University of Minnesota Medical School

The Robin Cook Backstory: Behind the Creation of the Medical Thriller Genre

Robin B. Cook, MD (1975) - Ophthalmologist & Bestselling Author

Introduction of the Distinguished Innovator Awardee

Joan W. Miller, MD (1989, 1991) - David Glendenning Cogan Professor of Ophthalmology and Chair, Department of Ophthalmology, Harvard Medical School; Chair of Ophthalmology, Mass Eye and Ear and Mass General Hospital; Ophthalmologist-inChief, Brigham and Women’s Hospital

2025 Distinguished Innovator Award Lecture

Crossing Barriers with a Vision on Genetic Medicine

Luk H. Vandenberghe, PhD - Associate Professor of Ophthalmology (Part-time), Harvard Medical School; Associate Director, Harvard Ophthalmology Ocular Genomics Institute, Grousbeck Family Chair in Gene Therapy, and Director, Grousbeck Gene Therapy Center, Mass Eye and Ear

Closing Remarks & 2026 Save the Date

Joseph F. Rizzo III, MD (1986) and Joseph B. Ciolino, MD (2004, 2009) - Meeting Co-Chairs

6:00-7:30 pm

Trainee Poster Contest & Reception

TRAINEE POSTER CONTEST -

Candidate: Darin Chhing

Poster #: 1

Analysis of Rho-Kinase Inhibitors on In Vitro Patient-Derived Model of Proliferative Vitreoretinopathy

Darin Chhing, Michael O’Hare

Purpose: The purpose of this study was to evaluate the effects of a novel Rho-Kinase inhibitor on cell proliferation and cytotoxicity in a cell line derived from patients with proliferative vitreoretinopathy (C-PVR).

Methods: C-PVR cells were treated with three Rho-Kinase inhibitors, Netarsudil, Fasudil, and Drug X (a novel Rho-Kinase inhibitor), and analyzed using the CyQuant cell proliferation assay and CytoTox 96 assay at 24, 48, and 72-hour time points.

Results: Netarsudil demonstrated significant reduction in cell proliferation between 2.5-5 uM at 48 and 72-hour time points, and reduced percent cytotoxicity below 2.5 uM. Fasudil did not feature significant reduction in cell proliferation above 1uM. Drug X exhibited significant reduction in cell proliferation between 2.5-10 uM at 48 and 72-hour time points, and reduced percent cytotoxicity below 10 uM across 72 hours of treatment.

Conclusions: A new Rho-Kinase inhibitor demonstrates effective reduction in cell proliferation with low cytotoxicity across a wider drug concentration and over longer duration in C-PVR cells compared to Netarsudil and Fasudil.

Candidate: Jie Liu

Poster #: 2

Epigenetic Adaptation Drives Monocyte Differentiation Into Microglialike Cells Upon Engraftment Into the Retina

Jie Liu, Fengyang Lei, Bin Yan, Tian Cao, Jyoti Sharma, Victor Correa, Lara Roach, Savvas Nicolaou, Kristen Pitts, James Chodosh, Daniel E. Maidana, Demetrios Vavvas, Milica A. Margeta, Raul Mostoslavsky, Eleftherios I. Paschalis

Purpose: The identification of specific markers for microglia has been a long-standing challenge. Recently, markers such as P2ry12, TMEM119, and Fcrls have been proposed as microglia-specific and widely used to explore microglial functions within various central nervous system (CNS) contexts. The specificity of these markers was based on the assumption that circulating monocytes retain their distinct signatures even after infiltrating the CNS. However, recent findings reveal that infiltrating monocytes can adopt microglia-like characteristics while maintaining a pro-inflammatory profile upon permanent engraftment in the retina. The current study is to investigate if infiltrating monocytes will acquire these proposed microglia markers in the retina after injury.

Methods: In this study, we utilize bone marrow chimeras, single-cell RNA sequencing, ATAC-seq, flow cytometry, and immunohistochemistry.

Results: This study demonstrateed that engrafted monocytes acquire expression of established microglia markers P2ry12, TMEM119, Fcrls and the pan-myeloid marker Iba1, which has been commonly mischaracterized as microglia-specific. These changes are accompanied by alterations in chromatin accessibility and shifts in chromatin binding motifs that are indicative of microglial identity. Moreover, we show that engrafted monocytes dynamically regulate the expression of CX3CR1, CCR2, Ly6C, and transcription factors PU.1, CTCF, RUNX, AP-1, CEBP, and IRF2, all of which are crucial for shaping microglial identity.

Conclusions: This study is the first to illustrate that engrafted monocytes in the retina undergo both epigenetic and transcriptional changes, enabling them to express microglia-like signatures. These findings highlight the need for future research to account for these changes when assessing the roles of monocytes and microglia in CNS pathology.

Candidate: Seyed MohamadMehdi Moshtaghion

Poster #: 3

A Small Molecule Therapy for Retinitis Pigmentosa: Addressing Autophagy and Ciliary Defects

Seyed MohamadMehdi Moshtaghion, Estefanía Caballano-Infantes, Laurie Clauzon, Álvaro Plaza Reyes, Risako Ido, Asahi Matsuoka, Andrea Garrido Gomes, Marta Torres Valcárcel, Berta De La Cerda Haynes, Juan C. Morales, Francisco Díaz-Corrales

Purpose: Retinitis pigmentosa (RP) is a genetically diverse retinal disease, with EYS mutations being a major cause of autosomal recessive RP (arRP). While EYS is traditionally linked to photoreceptor degeneration, its role in retinal pigment epithelial (RPE) cells remains unclear. Our group found that EYS mutations disrupt RPE function in patient derived iPSCs, suggesting a broader role for EYS beyond photoreceptors. This study investigates EYS localization in ARPE-19 cells, explores its impact on autophagy and ciliary dynamics in mutant RPE, and evaluates the therapeutic potential of Piceid Octanoate (PIC-OCT), a polyphenol derivative that enhances SIRT1 activity, a regulator of autophagy and ciliary function.

Methods: Subcellular localization of EYS in ARPE-19 cells was determined using immunofluorescence with antibodies against EYS and markers for organelles, cytoskeletal components, and intracellular vesicles. Patient-derived iPSCs with EYS mutation were differentiated into RPE cells to analyze autophagy (LC3-II/I conversion, p62 accumulation) and primary cilia morphology (immunofluorescence imaging). PIC-OCT was tested at concentrations of 100, 250, and 500 nM over 30 days on 50-day differentiated RPE cells to evaluate its effects on autophagy and ciliary dynamics. Statistical analyses compared treated and untreated groups.

Results: In ARPE-19 cells, EYS formed filamentous and vesicular structures near the nucleus. The filamentous structures colocalized with vimentin but not microtubules, actin filaments, or centrosomes. Vesicular structures partially colocalized with PROM1 and LAMP2, suggesting a role in vesicular trafficking. In EYS-mutant RPE cells, autophagy was impaired, evidenced by increased p62 levels and reduced LC3-II/I conversion after bafilomycin induction. Primary cilia were significantly shorter, with a 13.1% reduction in length (p = 0.001). Treatment with 250 nM PIC-OCT restored autophagy by reducing p62 levels, enhancing LC3-II/I conversion, and increased ciliary length by 5.2% (p = 0.03) compared to untreated cells.

Conclusions: EYS forms filamentous and vesicular structures associated with vimentin, endosomes, and lysosomes in ARPE-19 cells suggesting roles in intracellular organization and trafficking. EYS mutations impair autophagy and disrupt ciliary dynamics in iPSC-derived RPE cells, underlying RP pathogenesis. PIC-OCT effectively mitigates these deficits, demonstrating potential as a therapeutic approach for addressing autophagy and ciliary dysfunction in EYS-associated RP.

Candidate: Hannah Rana

Poster #: 4

Retinal Encoding Using a Difference of Gaussians Filter: Advancing Spiking Neural Networks for

Inspiring Retinal Prosthetics for Human Vision Restoration

Hannah Rana, Mohammad Eslami, Michael Morley, Nazlee Zebardast, Mengyu Wang, Tobias Elze

Purpose: This study advances the simulation of biologically plausible retinal encoding using a difference of Gaussians (DoG) filter that mimics the center-surround receptive fields of human retinal ganglion cells (RGCs), using the NE15-MNIST approach as an inspiring basis. This framework aims to advance spiking neural networks (SNNs) modeling, with direct applicability to retinal prostheses

Methods: A DoG filter was designed to simulate ON-center and OFF-surround responses, modeling human RGCs. The filter parameters included 2σ center and 5σ surround standard deviations, which were subtracted to create the DoG filter, and matrix dimensions of 11x11 pixels. A synthetic dataset simulating MNIST digits and the NE15-MNIST dataset’s Poisson-encoded subset were used as inputs to the encoding framework. The encoded spike trains were mapped to a simulated electrode grid to represent prosthetic stimulation patterns. The outputs were assessed for sparsity (percentage of inactive pixels; electrodes), and spike count, which are key metrics for evaluating encoding efficiency and biological fidelity in neuromorphic systems. The visualization of encoded spike trains highlighted spatial and temporal patterns, which are crucial for understanding information processing in retinal circuitry.

Results: The DoG filter successfully generated biologically realistic spike patterns, encoding synthetic and NE15-MNIST images. The sparsity averaged 78.5% (±3.2), indicating energy-efficient encoding, and the spike counts ranged between 1300-2100 spikes per image, capturing the essential features of the input stimuli. The visualization demonstrated clear ON-OFF response patterns that mimic retinal ganglion cell activity.

Conclusions: This study demonstrates the feasibility of using a biologically inspired encoding framework to simulate human retinal activity. The DoG filter effectively translates visual stimuli into spike-based representations that preserve spatial and temporal features. These results highlight the applicability of the approach to retinal prostheses, where mimicking natural ganglion cell activity is critical. Future work will focus on validating this model using live retinal spiking data, extending the model to process dynamic inputs of moving stimuli, and ultimately, developing deep learning models to generate predictive outputs to visual stimuli.

Candidate: Gustavo Sakuno

Poster #: 5

Targeting TMED9 to Attenuate Retinal Degeneration in a Mouse Model of Retinitis Pigmentosa

Gustavo Sakuno, Edward Ryan Collantes, Dimitrios Ntentakis, Toshio Narimatsu, Tian Cao, Juan Lorenzo B. Pablo, Anna Greka, Demetrios G. Vavvas

Purpose: Proteinopathies causing endoplasmic reticulum (ER) stress drive degeneration in many inherited diseases, including Retinitis Pigmentosa (RP), which lacks effective treatments. Transmembrane Emp24 domain-containing protein 9 (TMED9) is a key ER-Golgi cargo receptor implicated in protein trafficking. TMED9 knockout promotes lysosomal degradation of misfolded proteins, thereby alleviating ER stress a mechanism shown to be effective in other proteinopathies [e.g., mucin kidney disease (MUC-1), uromodulin kidney disease (UMOD)]. We hypothesized that conditional TMED9 knockout would mitigate photoreceptor degeneration caused by proteotoxicity in the P23H rhodopsin mouse model of RP.

Methods: To assess TMED9 knockout effects on retinal structure and function, C57BL/6J mice were used. These mice were heterozygous for the P23H rhodopsin mutation, homozygous for a TMED9 floxed allele, and either possessed (Cre+) or lacked (Cre–) a tamoxifen-inducible CAG-Cre-ESR transgene. A total of 25 mice (50 eyes) were analyzed. The knockout group comprised 14 Cre+ mice (10 male, 4 female), and controls were 11 Cre– littermates (5 male, 6 female). To induce recombination selectively in Cre+ mice, tamoxifen was administered intraperitoneally on postnatal days 21, 23, and 25; Cre– mice received tamoxifen to ensure consistent handling. Photoreceptors were evaluated at postnatal days 28, 35, and 42 (P28, P35, and P42) structurally by outer nuclear layer (ONL) thickness via optical coherence tomography (OCT), and functionally by a-wave scotopic electroretinography (ERG). Linear mixed-effects models (LMMs) were used for each outcome variable ONL thickness (µm), ERG scotopic a-wave amplitude (µV), and their fold-change from P28 baseline values (ratio) to control for inter-animal baseline variation with genotype, timepoint, and sex as fixed effects. The models included a random intercept for each mouse to account for the non-independence of measurements taken from both eyes of the same animal. Statistical models included sex as a covariate to adjust for group imbalance, and Bonferroni-corrected pairwise comparisons were performed at P35 and P42.

Results: At P42, Cre+ mice had greater ONL thickness (32.71 vs 30.14 µm; mean difference +2.58, 95% CI 0.35–4.81; Bonferroni-adj. p=0.047) and higher ERG a-wave ratio (2.07 vs 0.83; mean difference 1.25, 95% CI 0.44–2.05; Bonferroni-adj. p<0.01) than controls. No significant difference was found for ERG amplitude or ONL ratio at P42. However, LMMs assessing changes over time revealed significant genotype-by-timepoint interactions between P28 and P42 for ONL ratio, ERG amplitude and ERG ratio (all p < 0.01), with better outcomes for Cre+ groups over time. No sex differences were detected. These findings indicate superior preservation of photoreceptors in Cre+ mice at later stages.

Conclusions: Conditional TMED9 knockout significantly preserves retinal structure and function in the P23H model of RP. Our findings validate TMED9 as a promising therapeutic target for mitigating retinal degeneration caused by misfolded rhodopsin. Considering its protein trafficking role, TMED9 modulation (e.g., via gene editing or pharmacologically) warrants investigation as a potential treatment for other ER stress-driven diseases.

Candidate: Maryam Shayan

Poster #: 6

Effects of Vitamin D Supplementation in an Experimental Chronic Autoimmune Uveitis Model

Maryam Shayan, Katayoon Forouzanfar, John H. Kempen, Lucia Sobrin, Yihe Chen

Purpose: The incidence and severity of uveitis have been associated with low blood Vitamin D levels. The present study aims to investigate the impact of Vitamin D supplementation on the progression and immunopathology in our established mouse model of chronic autoimmune uveitis (CAU).

Methods: Two groups of C57BL/6J mice were studied: one group received a standard chew diet with regular vitamin D (2.3 IU/g, LabDiet® 5053) serving as the control group, while the other received a high-vitamin D diet (7.5 IU/g, LabDiet® modified 5053) for three weeks prior to CAU induction. CAU was induced via immunization with interphotoreceptor retinoid-binding protein (IRBP) peptides emulsified in complete Freund's adjuvant, followed by Bordetella pertussis toxin injection. Disease progression was monitored biweekly for 12 weeks using fundoscopy, optical coherence tomography (OCT), and electroretinography (ERG). The two groups of mice continued receiving standard or high-vitamin D diet respectively during the 12 weeks of follow up. At week 12, mice were euthanized and tissues including the retina, cervical lymph nodes, inguinal lymph nodes, and spleen, were collected for flow cytometric analysis of the pathogenic memory Th17 cells.

Results: Preventive vitamin D supplementation resulted in reduced disease severity as evidenced by significantly lower fundoscopic disease scores than the control group at weeks 2 (0.1±0.1 vs 0.8±0.2), 4 (0.4±0.2 vs 1±0), and 6 (0.6±0.2 vs 1±0). This effect persisted with gradually reduced differences until week 12 (0.7±0.2 vs 1±0, p=0.08). OCT analysis at week 12 demonstrated reduced retinal thickness in the control group, consistent with retinal atrophy observed in chronic uveitis, while the vitamin D-supplemented group showed well-preserved retinal thickness (change of retinal thickness from baseline: 0.7±0.9µm vs -3.9±1.1µm). ERG recordings demonstrated similarly well-preserved amplitudes of both dark-adapted and light-adapted aand b-waves in the vitamin D-supplemented group in contrast to the reduced responses in the control group throughout the 12 weeks of observation period, although the differences between the groups were not statistically significant. Flow cytometry analysis revealed reduced frequencies of memory Th17 cells in the retina, cervical lymph nodes and spleen in the vitamin D-supplemented group compared to the control group.

Conclusions: Vitamin D supplementation has the potential to reduce the severity and progression of CAU by modulating the underlying immune response, highlighting the promise of oral vitamin D as an effective therapeutic strategy for chronic uveitis.

Candidate: Anil Upreti

Poster #: 7

Transcriptomic Landscape of Topoisomerase I Inhibition in Retinal Fibrosis

Anil Upreti, Jeysson Sanchez-Suarez, Karim W. Barake, Rose Lin, Elizabeth E. Rossin, Michael O’Hare, Leo A. Kim

Purpose: Proliferative vitreoretinopathy (PVR) is a serious complication of retinal detachment and ocular trauma, characterized by the formation of contractile membranes within the vitreous cavity and on the retinal surface. Currently, there are no FDA-approved pharmacologic treatments for PVR, with surgery being the only available option. This study investigates the potential of topotecan (TOP), a topoisomerase I inhibitor, as a therapeutic agent using an in vitro model of PVR.

Methods: Single-cell RNA sequencing (scRNA-seq) analysis of patient-derived PVR membranes revealed elevated expression of TOP1 in PVR tissues. Functional validation was performed using a primary cell culture model derived from PVR patients (C-PVR cells). These cells were treated with increasing concentrations of topotecan (TOP; 2.5–160 nM), and cell proliferation and cytotoxicity were assessed using CyQuant and LDH assays, respectively. To investigate pathway-level changes, bulk RNA sequencing (RNA-seq) was conducted on C-PVR cells cultured for 48 hours under four conditions: vehicle control, TOP (100 nM), TGFβ2 (5 ng/mL), and a combination of TOP and TGFβ2.

Results: TOP treatment significantly inhibited C-PVR cell proliferation in a dose-dependent manner, with effects observed at concentrations as low as 10 nM (p < 0.05). Importantly, this antiproliferative effect occurred without inducing cell death, as LDH levels remained undetectable up to 72 hours post-treatment. Bulk RNAseq analysis identified significantly differentially expressed genes (log2FC > 1.5, padj < 0.05) across pairwise comparisons. Hierarchical clustering of DEGs revealed treatment-specific transcriptional profiles. Notably, topotecan-treated cells showed marked enrichment of the p53 signaling pathway, along with pathways related to apoptotic signaling, cell cycle regulation, VEGF signaling, and extracellular matrix remodeling.

Conclusions: These findings support the potential of topotecan as a promising therapeutic candidate for modulating cellular proliferation and fibrotic signaling in PVR. Together, our data highlight the therapeutic relevance of targeting topoisomerase I and provide mechanistic insight into transcriptomic alterations in response to topotecan treatment in PVR.

Candidate: Krupa Sourirajan

Poster #: 8

Metabolomic Profiling of Normal Human Retinal and Choroid Tissue

Krupa Sourirajan, Ines Lains, Kevin Mendez, Haemin Kang, Roshni Bhat, Augustine Bannerman, David M. Wu, Ivana K. Kim, John B. Miller, Demetrios G. Vavvas, Jessica Lasky-Su, Joan W. Miller, Deeba Husain

Purpose: Metabolomics, the study of metabolites, has been successfully applied to the study of many retinal diseases. Prior work has mostly been conducted using plasma and serum samples. To our knowledge, no prior characterizations of metabolomic profiles of human retinal tissue have been reported. This is crucial to understand how biofluid changes relate to in situ findings. This study aimed to characterize the normal metabolomic profile of donor human retinal tissue samples.

Methods: Prospectively designed cross-sectional study including human donor eyes (n=14) above the age of 50 without any retinal disease that were enucleated within six hours post-mortem. After the anterior segment and corneal button were removed, vitreous was collected and the remaining eye cup was transferred to a sterile saline petri dish. A 6 mm macular punch, followed by a periphery punch were then obtained. The remaining retina was separated. For these samples (macula, periphery and remaining retina), the neurosensory retina was then separated from retinal pigment epithelium (RPE)/choroid. Color fundus photographs were taken both prior to and following the removal of the neural retina. These images were reviewed to ensure the absence of retinal diseases. Serum samples were collected for the same donors. All samples were snap frozen and stored at -80C degrees in sterile vials and were shipped frozen. Non-targeted mass spectrometry analysis was conducted by Metabolon Inc.

Results: A total of 659 metabolites were identified in the retina/choroid tissue samples. Most of them were lipids (53%) and amino acids (21%). The remaining metabolites belonged to nucleotide (7%), carbohydrate (6%), cofactors and vitamins (6%), xenobiotics (3%), peptide (2%), and energy (2%) pathways. The same type of tissue (neurosensory vs RPE/choroid) had a similar metabolomic profile irrespective of location (macula vs periphery). When looking at neurosensory retina vs RPE, the most unique metabolites were 8 dinucleotides to RPE, and 5 lyphospholipids for neurosensory retina. There were 513 metabolites (46.5%) overlapping between the retina/choroid tissue and serum.

Conclusions: To our knowledge, this study presents the first assessment of metabolomic profiles of human retina/choroid tissue. Most of the identified circulating metabolites were also seen in the retina/choroid tissue, thus reaffirming their potential use as biomarkers for retinal disease. Understanding the relationship between circulatory and eye-specific metabolites is crucial for enabling future translational medicine research.

Candidate: Said Arevalo Alquichire

Poster #: 9

Targeting RUNX1 Reverses Vascular Pathology in Model of NOTCH3 Deficiency

Said Arevalo-Alquichire, Rafael Posada-Duque, Michael O’Hare, Juliana Gonzalez-Perez, Andres Muriel, Timothy Vanderleest, William P. Miller, Leo A. Kim, Joseph F. Arboleda-Velasquez

Purpose: NOTCH3 is a relevant pathway in cell development and differentiation of vessels. NOTH3 defiency have been implicated in conditions like diabetic retinopathy where the loss of pericytes is a hallmark. Moreover, retinal vascular abnormalities, such as cotton wool spots, nerve fiber loss, elevated outer diameters, and reduced vascular density, have been identified in patients carrying NOTCH3 mutations. In this work, we identified and validate novel markers, via single-cell RNA sequencing, that mediate the loss of pericytes and vascular pathology in retinal diseases associated with a breakdown in the NOTCH signaling.

Methods: We examined the retinal vascular pathology of mouse models carrying NOTCH3 mutations, and we explored the preclinical efficacy of targeting new biomarkers identified from the analysis single RNA sequencing of Fibrovascular membranes derived from PDR and models of carrying NOTCH3 mutations . Animals carrying a NOTCH3 mutation (C445R) that previously described loss of pericytes were injected with a small molecule inhibitor and the retinas were assessed via fundus fluorscence angiography and retina flat mount staining.

Results: We identified that the transcription factor RUNX1 was upregulated in human tissue and models of NOTCH3 deficiency. Subsequently, we used mouse models for NOTCH3 mutations to assess the efficacy of RUNX1 inhibition as a therapeutic approach for treating vascular pathology. We administered a small molecule inhibitor of RUNX1 called Ro24-7429 via intraperitoneal injection for one month and assessed the retinal vasculature using fundus fluorescein angiography and immunofluorescence. We observed that mice carrying the C455R mutation developed vascular retinal pathology at the age of six months , and the pathology progressed with time. Moreover, we found that the inhibition of RUNX1 curbed pathology development, induced recovery of the tone of the vessels, and increased the levels of ɑSMA in the retinal vessels.

Conclusions: We conclude that targeting the transcription factor RUNX1 could be a novel therapeutic approach for treating vascular pathology associated with NOTCH3 deficiency.

Candidate: Roshni Bhat

Poster #: 10

Retinal Tissue Metabolomics: An Analysis of AMD Metabolomic Profiles

Roshni Bhat, Ines Lains, Kevin Mendez, Haemin Kang, Krupa Sourirajan, Augustine Bannerman, David M. Wu, Ivana K. Kim, John B. Miller, Demetrios G. Vavvas, Jessica Lasky-Su, Joan W. Miller, Deeba Husain

Purpose: Metabolomics, the study of small molecules (<1 kD) in our body fluids and tissues, provides insight into the pathophysiology of multifactorial diseases, like age-related macular degeneration (AMD). Prior work has focused on plasma and urine metabolomic associations with AMD. However, metabolomics of retinal samples remains largely unexplored, and it remains unclear how biofluid metabolomic profiles relate to in situ changes in this disease. This work aimed to compare human retinal tissue metabolomic profiles of donor patients with AMD vs controls.

Methods: Cross-sectional study including 38 post-mortem (<6hrs) human donor eyes of subjects over age 50 (25 AMD, 13 control). After the anterior segment and corneal button were removed, vitreous was collected. The remaining eye cup was transferred to a sterile saline petri dish and color fundus photographs were taken, which were then reviewed to assess for the presence of AMD. Two 6 mm macular punches were obtained; from the macula and from the periphery and the remaining retina processed. For each sample (macula, periphery and remaining retina), the neurosensory retina was separated from retinal pigment epithelium (RPE)/choroid. All samples were snap frozen and stored at -80C degrees in sterile vials and kept frozen. Nontargeted mass spectrometry analysis was conducted by Metabolon Inc. Multivariate logistic regression models, accounting for age and gender, were used for analysis with statistical significance determined by p < 0.05.

Results: In macula neurosensory retinal tissue, 45/559 metabolites were found to have a negative association with AMD. Most of them were lipids, (n=35, 77.7%), including n= 7 lysophospholipids, n=11 phosphatidylcholines, n=3 phosphatidylethanolamines and n=3 phosphatidylinositol. Additionally, there were 6 metabolites significantly associated with presence of AMD in macular RPE/choroid tissue, again most (n=4, 66.67%) from lipid super-pathways.

Conclusions: This is the first study investigating metabolomic profiles of human retinal tissue in AMD. Our findings are in agreement with previous results from our group using plasma samples, demonstrating differences in metabolomic profiles of AMD patients vs controls in phosphatidylcholines and phosphatidylethanolamines among other lipids. Taken together, these findings validate that metabolomics can lead to better understanding of the pathogenesis of this disease.

Candidate: Meenakshi Maurya

Poster #: 11

Genetic Deletion of REV-ERBα Regulates Retinal Mitochondrial Metabolism

and Age-related Synaptic Remodeling in Mice

Meenakshi Maurya, Chi-Hsiu Liu, Kiran Bora, Neetu Kushwah, Madeline C. Pavlovich, Zhongjie Fu, James D. Akula, Jing Chen

Purpose: Synaptic remodeling in the outer retina occurs in normal aging and is exacerbated in age-related macular degeneration (AMD), associated with compromised neuronal function. The age-related mitochondrial decline is a key feature in many metabolic and neurological diseases in both humans and mice and may underlie retinal synaptic alterations during aging. REV-ERBα, a redox-sensitive nuclear receptor, regulates circadian rhythm, lipid and glucose metabolism, and mitochondrial biogenesis. The present study explored the role of REV-ERBα in the maintenance of retinal mitochondrial function and synaptic integrity in the outer retina during aging using systemic (Rev-erbα-/-) and rod-specific (Rho-icre:Rev-erbαfl/fl) knockout mice.

Methods: Rev-erbα-/- and Rho-icre:Rev-erbαfl/fl mice with their respective age-matched controls (WT or flox) were generated. Genetic deletion of Rev-erbα in knockout retinas was confirmed using IHC, RT-qPCR, and immunoblotting. Ectopic photoreceptor synapses were visualized and quantified in aged Rev-erbα-/- and Rhoicre:Rev-erbαfl/fl retinas with age-matched controls. Visual function was assessed using scotopic ERG. Retinal mitochondrial activity was measured with a Seahorse metabolic analyzer. Potential target metabolic genes of Rev-erbα were assessed using qPCR and immunoblotting. In 661w photoreceptor cells, mitochondrial network dynamics were evaluated using MitoTracker in REV-ERBα knockdown ShRev-erbα treatment vs. ShControl.

Results: REV-ERBα was localized in most retinal neurons including rods and bipolar cells with IHC. REVERBα deletion results in acceleration in ectopic photoreceptor synapses with aberrant rod bipolar cell dendritic outgrowth into the outer nuclear layer. In ~1-year-old mouse retinas, both the number and the distance of photoreceptor synapse retraction were increased in both Rev-erbα-/- and Rho-icre:Rev-erbαfl/fl retinas compared to age-matched controls (n=8 images, 3 mice/ group, p<0.001), that were accompanied by a substantial increase in the number and length of ectopic rod-bipolar dendrite sprouts (p<0.001). Rev-erbα-/shows significant impairment of visual function in ~1-year-old mice, with reduction in the amplitudes of photoreceptor-specific a-wave (p=0.005) and rod-bipolar specific b-wave (p=0.007) in Rev-erbα-/- vs WT controls. Rho-icre:Rev-erbαfl/fl mice also had compromised visual function with a decrease in amplitudes of a wave (p=0.03) and b-wave (p=0.009) relative to flox controls (n=6 mice/group). Rev-erbα-/- retinas demonstrated a substantial decrease of mitochondrial activity measured by oxygen consumption rate using a Seahorse analyzer. REV-ERBα deficiency led to a significant decrease of AMPK regulator LKB1 in both mRNA and protein levels in both Rev-erbα-/- and Rho-icre:Rev-erbαfl/fl retinas relative to their controls.

Upregulation in phospho-DRP1(s616) and mitochondrial fission factor, a receptor for fission protein DRP1 confirms that REV-ERBα systemic deletion accelerates mitochondrial fission and disintegration. In 661W cells, the ShRev-erbα treated cells show significantly decreased mitochondrial density and network footprint compared to ShControl.

Conclusions: Our findings suggest that both Rev-erbα-/- and Rho-icre:Rev-erbαfl/fl accelerates age-related photoreceptor synaptic remodeling, potentially through regulation of mitochondrial metabolism via the LKB1AMPK axis, indicating its photoreceptor-intrinsic role in mediating the effects of aging on mitochondrial integrity.

Candidate: Aruvi Vijikumar

Poster #: 12

Multiomics

Analysis of Endothelial-Mesenchymal Transition via the TNFα-RUNX1 Pathway

Aruvi Vijikumar, William P. Miller, Anil Upreti, Yvonne Adu-Rutledge, Said Arevalo-Alquichire, Audrey L. Gunawan , Elizabeth J. Rossin, Michael O’Hare, Leo A. Kim

Purpose: Continuous administration of anti-VEGF therapy has been shown to induce fibrosis in Proliferative diabetic retinopathy (PDR), leading to the contraction and progression of fibrovascular membranes (FVM). Ocular fibrosis is a complex biological process that is a leading cause of visual impairment and blindness worldwide. This underscores the unmet clinical need to develop effective anti-fibrotic therapies that can be administered in conjunction with or following anti-VEGF agents, especially in individuals at high risk of scarring, to preserve visual function. Our previous studies have identified the Runt-related transcription factor 1 as a critical regulator of retinal fibrosis, with TNFα inducing RUNX1-mediated endothelial-to-mesenchymal transition. The aim of this study is to further elucidate the role of RUNX1 as a potential therapeutic target for mitigating ocular fibrosis.

Methods: Human retinal endothelial cells (HMRECs) treated with TNFα (5ng/ml) for 72H were used to identifiy the molecular mechanisms of TNFα-induced RUNX1 regulating the progression of Endo-MT by performing a high-throughput single-cell multiome analysis, integrating ATAC-seq and RNA-seq data. Effects of RUNX1 inhibition was assessed in Kimba (TrVEGF029) mouse model of genetically induced VEGF. Animals were evaluated by OCT, and fluorescein angiography.

Results: An integrative analysis of transcriptomics and chromatin accessibility data revealed that TNFα treatment in HMRECs induced a robust mesenchymal transition and fibrosis signature. A dynamic shift in the chromatin landscape was observed in ECs following TNFα stimulation, suggesting a potential role for RUNX1 in regulating the pro-fibrotic transition. Furthermore, treatment with RUNX1 inhibitor significantly reduced retinal neovascularization and improved retinal pathologies relative to untreated controls in kimba model.

Conclusions: Our results indicate that RUNX1 is a major driver of the profibrotic phenotype in retinal endothelial cells. These findings suggest that targeting RUNX1 may represent an important therapeutic approach to curb the proliferation of fibrotic plaques and preserving visual function.

Candidate: Shuai Wang

Poster #: 13

Nrf2 Gene Therapy Preserves Retinal and Choroidal Vasculature in an Oxidative

Stress-Induced Model of AMD

Shuai Wang, Sophia Rong Zhao, Apolonia Gardner, Adam Daniels, Constance Cepko

Purpose: Oxidative stress is a key contributor to dry age-related macular degeneration (AMD). While Nrf2 gene therapy has shown neuroprotective effects on the RPE and photoreceptors, its role in preserving retinal and choroidal vasculature is unclear. This study investigates whether AAV8-BEST1-driven Nrf2 overexpression can prevent retinal vascular leakage and choroidal atrophy in an oxidative stress-induced AMD mouse model.

Methods: AAV8-BEST1-Nrf2 was delivered via subretinal injection into P0 neonatal or adult C57BL/6J mice. Sodium iodate (NaIO₃, 75mg/kg, IP) was administered either 6–8 weeks post-pup injection or 1 week postadult injection to induce oxidative stress. Vascular leakage was assessed by fluorescein angiography (FA) at 1 day, 1 week, and 4 weeks post-NaIO₃. Retinal and choroidal vascular structures were analyzed by isolectin B4 staining on retinal flatmounts and podocalyxin staining on RPE/choroid flatmounts. Additionally, RPE/choroid explants from 8-week-old mice that received P0 Nrf2 injections were cultured for 24 hours, and the supernatant was collected. This conditioned medium was then injected subretinally into adult mice, which were subsequently challenged with NaIO₃ after 2 days. FA was performed at 1 day, 1 week, and 4 weeks postNaIO₃. Eyes were harvested at 4 weeks for vascular structure evaluation.

Results: AAV8-BEST1-Nrf2 significantly reduced retinal vascular leakage and preserved choroidal vasculature in both neonatal and adult-injected groups after S.I. Structural integrity of the deep retinal and choroidal vasculature was retained at 4 weeks post-oxidative stress. Furthermore, conditioned media from Nrf2-treated RPE/choroid explants conferred similar vascular protection in recipient mice, reducing leakage and maintaining vessel architecture, indicating a secreted factor-mediated effect.

Conclusions: Nrf2 gene therapy not only protects the RPE but also preserves the integrity of retinal and choroidal vasculature under oxidative stress. Subretinal delivery of AAV8-BEST1-Nrf2 prevented vascular leakage and structural degeneration in a NaIO₃-induced AMD model. The efficacy of the supernatant suggests that Nrf2 may promote the release of soluble protective factors. These findings support a novel therapeutic strategy for dry AMD targeting both neural and vascular components of the disease.

Candidate: Helia Ashourizadeh

Poster #: 14

Impact of Cataract Surgery on the Risk of Conversion From Dry to Neovascular Age-related

Macular Degeneration in the IRIS® Registry

Helia Ashourizadeh, Joshua B. Gilbert, Nicole Grinspan, William C. Kearney, Connor Ross, Tobias Elze, Joan W. Miller, Alice C. Lorch, Grayson W. Armstrong, IRIS® Registry Analytic Center Consortium

Purpose: To evaluate conversion rate from dry age-related macular degeneration (AMD) to neovascular AMD (nvAMD) following cataract surgery and identify associated risk factors.

Methods: A 7-year retrospective time-to-event study examined dry AMD patients who underwent cataract surgery (treatment) in the IRIS® Registry (Intelligent REsearch in Sight). We conducted a within-person (treated vs. untreated fellow eye) and a full-sample analysis using Kaplan-Meier survival and proportional hazards models.

Results: Full sample included 241,732 eyes (mean age 76 ± 7 years, 61% female). At 7 years, conversionfree survival was 68% in operated eyes versus 32% in unoperated eyes (N=25,111 eyes). In within-person analysis, operated eyes had significantly lower conversion risk (HR=0.49, P<0.001), controlling for pre-op dry AMD stage, visual acuity, and demographics. In full sample analysis, worse baseline vision, older age, female sex, smoking, advanced AMD and White race (P<0.001) increased the risk.

Conclusions: Cataract surgery was associated with a lower conversion rate to nvAMD. Advanced dry AMD, older age, worse baseline vision, female sex, White race and smoking increased conversion risk.

Candidate: Xinyi Ding

Poster #: 15

Expanded Field OCT Angiography Biomarkers for Predicting Clinically Significant Outcomes in Non-Proliferative Diabetic Retinopathy

Xinyi Ding, Francesco Romano, Itika Garg, Jenny Gan, Filippos Vingopoulos, Demetrios G. Vavvas, Deeba Husain, Nimesh A. Patel, Leo A. Kim, John B. Miller

Purpose: To evaluate the utility of extended field swept-source Optical Coherence Tomography Angiography (SS-OCTA) imaging biomarkers in predicting the occurrence of clinically significant outcomes in eyes with NonProliferative Diabetic Retinopathy (NPDR).

Methods: Single-center clinical study. Eighty-eight eyes with NPDR from 57 participants (median age: 64.0 years; mean duration of diabetes: 15.8 years) with at least 2 consecutive SS-OCTA scans over a follow-up period of at least 6 months were included. The presence of intraretinal microvascular abnormalities (IRMAs) at baseline and the stability of IRMAs during follow-up period on 12 ×12-mm angiograms were evaluated. Baseline nonperfusion ischemia index (ISI) and other SS-OCTA metrics were calculated on FIJI and ARI Network. Significant clinical outcomes were defined as occurrence of one or more of the following events at the last available clinical visit:1. significant DR progression (2-step DR progression or progression to proliferative DR (PDR)); 2) development of new center-involving diabetic macular edema (CI-DME); and 3) initiation of treatment with PRP or anti-VEGF injections during the follow-up period. Mixed-effects Cox regression models was used to explore these outcomes.

Results: Following a clinical follow-up period lasting 25.1 ±10.8 months, we observed significant clinical outcomes in 17 eyes (19.3%). Among these, 7 eyes (8.0%) experienced significant progression, and 4 eyes (4.5%) developed CI-DME. Anti-VEGF injections were initiated in 15 eyes (17.0%), while PRP was initiated in 2 eyes (2.3%). Upon adjusting for age, the duration of DM, and prior Anti-VEGF treatments, our analysis revealed that non-stable IRMAs during follow-up periods and a higher ischemia index at baseline were significantly as- sociated with the occurrence of significant clinical out- comes with HRs of 3.88 (95% CI: 1.569.64; p = .004) and 1.05 (95% CI: 1.02-1.09; p = .004), respectively.

Conclusions: In conclusion, NPDR eyes with non-stable IRMAs over time and more ischemia at baseline are in higher risk of developing significant clinical outcomes. Our findings suggest that expanded field SS-OCTA may offer additional prognostic benefits for clinical DR staging and predicting high-risk patients.

Candidate: Selin Gumustop

Poster #: 16

Utilization of Ultra-Widefield OCT Angiography in Assessing NonPerfusion Areas and Their Correlation With Diabetic Retinopathy Severity and Predominantly Peripheral Lesions

Selin S. Gumustop, Xinyi Ding , Yi Stephanie Zhang, Ioanna Ploumi, Leiyu Wang ,Ying Zhu, Francesco Romano, Itika Garg, Chong Chen, Demetrios G. Vavvas, Deeba Husain, Joan W. Miller, Nimesh A. Patel, Leo A. Kim, John B. Miller

Purpose: To compare the non-perfusion areas (NPA) between two scan fields (12×12-mm vs 21×26-mm) using UWF-OCTA and to evaluate their correlation with the severity of diabetic retinopathy (DR) as well as predominantly peripheral lesions (PPL).

Methods: This cross-sectional study included patients with DR (non-proliferative [NPDR] and proliferative [PDR] without pan-photocoagulation) in at least one eye, aged > 18 years, and VA> 20/200. The study encompassed 101 eyes (62 patients), with 38 eyes (37.6%) classified as mild NPDR, 26 eyes moderate NPDR (25.7%), 15 eyes severe NPDR (14.9%) and 22 eyes PDR (21.8%) (All eyes were imaged with UWF-OCTA (DREAM OCT, Intalight) (12×12-mm and 21×26-mm scans). NPA were calculated using a semi-automatic algorithm as previously published on FIJI (Figure 1). The ischemic index (ISI) was determined by dividing the NPA by the total scan area. A subset of eyes with same-day ultra-widefield color fundus photography (UWF CFP) were assessed by 2 independent graders for the presence of and the number of fields with PPLsdefined as more than 50% of diabetic lesions subjectively residing in the extended field compared to the respective ETDRS field. Only 1 field with PPLs was required for the eye to be graded as having PPLs. The primary outcome was the comparison of ISI between the two scan sizes stratified for DR status using the Mann-Whitney U Test. Secondary outcomes used ordered logistic regression (adjusting for age and diabetes duration) and ROC analysis to evaluate the association between the ISI from different scan sizes and the severity of DR. A sub-analysis looked at the association of PPLs and retinal ischemia on the different scan sizes using a multivariable mixed-effects linear regression model.

Results: NPA and ISI were significantly greater in PDR than NPDR on 12×12-mm and 21×26-mm SS-OCTA and the NPA/ISI was significantly higher in the 21x26-mm field of view (p < 0.001 for all). Both the 12×12-mm ISI (OR: 1.26 [95% CI: 1.07-1.48, P = 0.006]) and the 21×26-mm ISI (OR: 1.20 [95% CI: 1.10-1.29, P < 0.001]) were significantly associated with increasing DR severity. The ROC analysis suggests comparable discriminative ability between both scan ISI at predicting DR severity with an AUC of 0.868 compared to 0.878 (p = 0.558). In the sub-analysis, 29 eyes (36.7%) had PPL. The presence of PPL was only significantly associated with ISI on the 21x26-mm scan (p=0.009).

Conclusions: The study demonstrated the effectiveness of the ISI from various angiographic scales in assessing DR severity. Additionally, there is evidence of associations between more peripheral NPAs and peripheral lesions (e.g., PPLs) which emphasizes the potential value of the ultra-widefield SS-OCTA in DR longitudinal monitoring and intervention guidance.

Candidate: Haemin Kang

Poster #: 17

Metabolic Signatures of Multiple Dietary Patterns and Their Association With Age-related Macular Degeneration

Haemin Kang, Huan Yun, Shuhua Xu, Kevin Mendez, Ines Lains, Krupa Sourirajan, Roshni Bhat, Rodrigo A. Alvarez, David Wu, Ivana K. Kim, John B. Miller, Demetrios G. Vavvas, Joan W. Miller, Jessica Lasky-Su, Liming Liang, Deeba Husain

Purpose: A metabolic signature, multi-metabolite profiles of dietary pattern, can be an objective and comprehensive way to evaluate adherence to diet, stratifying individuals based on dietary response and evaluating disease risk. This study aimed to validate metabolic signatures of various dietary patterns on our patient cohort, and to investigate their associations with age-related macular degeneration (AMD).

Methods: This cross-sectional study is a part of a prospective observational study on AMD biomarkers, including both participants with and without AMD. Metabolic profiling on fasting plasma samples was done by liquid chromatography-tandem mass spectrometry by Metabolon Inc. A validated food frequency questionnaire by Harvard School of Public Health was used to assess conventional dietary scores, and then metabolomic signatures were calculated. Eight dietary patterns were used including alternative Mediterranean diet [AMED], alternate healthy eating index [AHEI], anti-hypertensive diet [DASH], 3 plant-based diets [PDIs], inflammatory [EDIP] and insulinemic [EDIH] diets. To test the robustness and performance of the metabolic signatures, Pearson correlation coefficients were calculated between dietary scores and metabolic signatures. Multilevel mixed linear regression was used to estimate the association of dietary scores/metabolic signatures with risk of AMD, and covariates included age, sex, race, smoking, and body mass index.

Results: Among 1,008 eyes of 504 participants, 276 eyes (27.4%) had no evidence of AMD. Plasma metabolic signatures were robustly correlated with the corresponding dietary scores (r = 0.16 to 0.29; all P < 0.0001). Plasma metabolic signatures of AMED, DASH, healthful PDI and overall PDI were associated with lower risk of AMD (OR=0.84 to 0.88, all P < 0.016), whereas unhealthy PDI was associated with higher risk of AMD (OR=1.13, P=0.016). Corresponding dietary scores failed to show significant associations with risk of AMD (OR=0.94-0.99, P>0.282).

Conclusions: Plasma metabolic signatures could be validated in our AMD cohort, reflecting adherence and metabolic response to corresponding dietary patterns. Our results suggest that higher adherence to healthier diets were associated with reduced risk of AMD. Plasma metabolic signatures were significantly associated with risk of AMD, suggesting that plasma metabolic signatures can be more sensitive and objective way to assess dietary association with AMD than conventional dietary scores.

Candidate: Dimitrios Ntentakis

Poster #: 18

Conceptual Framework for Macular Telangiectasia Type 2

Pathogenesis: Insights From Human Histopathology

Dimitrios P. Ntentakis, Anastasia Maria Ntentaki, Eleni Delavogia, Gustavo Sakuno, Victor S.M.C Corrêa, Nikolaos Efstathiou, Mary E. Aronow, Emily Y. Chew, Joan W. Miller, Demetrios G. Vavvas

Purpose: To propose a conceptual framework for the predisposing factors and pathophysiologic progression of Macular Telangiectasia type 2 (MacTel), a rare neurodegenerative disorder with unclear pathogenesis and no definitive treatment.

Methods: A systematic review of histopathologic studies on MacTel donor eyes, published through December 31, 2024, was conducted. Key parameters evaluated included tissue processing techniques, post-mortem fixation intervals, histopathologic analysis types, premortem MacTel diagnosis confirmation, disease status at the last ophthalmologic evaluation, control specimen inclusion, and retinal section topography. Consistently reported histopathologic features in validated MacTel cases were identified and analyzed for clinical and histopathologic specificity, alongside their genetic, metabolic, and anatomic associations from existing literature. Donor demographics, co-existing ophthalmologic conditions, and systemic comorbidities were integrated to formulate a hypothesis for MacTel pathogenesis.

Results: Ten studies were reviewed, identifying three consistent findings in confirmed MacTel donor eyes: perifoveal macular pigment depletion, Müller cell loss corresponding to pigment depletion, and selective temporal perifoveal involvement. These findings were highly specific to MacTel. Müller cell degeneration likely initiates the disease, followed by photoreceptor atrophy. A genetic predisposition to retinal metabolic dysfunction, particularly impaired serine synthesis, may be exacerbated by ischemic stress, potentially linked to vascular insufficiency. The temporal perifovea's distinct embryologic and perfusion characteristics may explain its selective involvement. All donors exhibited ischemia-related risk factors, supporting a role of retinal hypo-perfusion trigger contributing to MacTel pathogenesis.

Conclusions: Müller cell degeneration, driven by genetic predisposition and ischemic stressors, likely initiates MacTel. Regardless of the specific genetic defect, ischemic stressors later in life exacerbate dysfunctional serine synthesis, amplifying retinal damage. Our conceptual framework, derived from rigorous analysis of consistent histopathologic findings, could offer a model for understanding rare retinal diseases and guiding hypothesis-driven research.

Candidate: Ioanna Ploumi

Poster #: 19

Associations Between Predominant Peripheral Lesions and Systemic Complications of Diabetes Mellitus

Ioanna Ploumi, Xinyi Ding, Francesco Romano, Jenny Gan, Edward Lu, Ying Cui, Krystal Phu, Ying Zhu, Kayla Nodecker, Shivesh Shah, Dimitrios Ntentakis, Itika Garg, Filippos Vingopoulos, Demetrios Vavvas, Deeba Husain, Leo Kim, Nimesh Patel, John Miller

Purpose: The presence of PPL, defined as more extensive diabetic retinopathy (DR) lesions in any one peripheral field compared to its corresponding ETDRS field, may indicate a more advanced DR severity and identify eyes at higher risk for future DR worsening independent of their baseline DR Severity Scale (DRSS) score. However, the relationship between PPL presence and the occurrence of other DM-related complications has not been fully explored. Identifying diabetic patients who are at high risk for microvascular and macrovascular complications could enhance the multispecialty care provided to these individuals.

Methods: This cross-sectional study included 182 eyes from 100 diabetic patients who were imaged using ultra-widefield color fundus photographs (UWF-CFP; California, Optos plc., Dunfermline, UK). UWF-CFP were assessed by 2 independent graders for presence and extent of PPL, defined as over 50% of diabetic lesions localized in the extended field compared to its respective ETDRS field. Only one field with PPL was required for the eye to be graded as having PPL. Comprehensive demographic and clinical data were collected from electronic medical records including diabetes data and history of DM-related complications. Generalized estimating equations adjusted for age, diabetes duration, HbA1c and smoking were used to determine the association between presence and extent of PPL and systemic complications of DM.

Results: PPL were identified in 69 out of 182 eyes [37.9%]. PPL presence was significantly associated with peripheral arterial disease (OR=11.36, p=0.033), coronary artery disease (OR=5.86, p<0.001), stroke (OR=10.11, p=0.003), diabetic nephropathy (OR=2.97, p=0.016), and decreased estimated glomerular filtration rate (β = -12.66, p = 0.037). Similarly, the extent of PPL was significantly associated with peripheral arterial disease (OR=1.63, p=0.036), coronary artery disease (OR=1.82, p=0.001), and stroke (OR=1.99, p=0.007). No significant associations were found for the presence or extent of PPL with diabetic neuropathy and diabetic foot.

Conclusions: The presence and extent of PPL are associated with both macrovascular and microvascular complications of DM. Early detection of PPL using UWF-CFP may aid in timely management of these severe and potentially fatal complications. Future prospective studies will be important to validate the associations identified in this study.

Candidate: Ying Zhu

Poster #: 20

Influence of Anti-VEGF Injections in Longitudinal Changes in Vascular Metrics

Measured by Optical Coherence Tomography Angiography in Age Related Macular Degeneration

Ying Zhu, Selin S. Gumustop, Leiyu Wang, Aurora Tong, Xinyi Ding, Ioanna Ploumi, Francesco Romano, Chong Chen, Kayla N. Nodecker, Shivesh H. Shah, Demetrios G. Vavvas, Deeba Husain, Joan W. Miller, Nimesh A. Patel, Leo A. Kim, David M. Wu, John B. Miller

Purpose: To investigate the possible influence of repeated anti-VEGF injections on vascular metrics measured by optical coherence tomography angiography (OCTA) in patients with age related macular degeneration (AMD).

Methods: This retrospective longitudinal study included AMD patients with a follow-up time of at least 18 months in a single tertiary medical center from 2019 to 2024. Swept-source OCTA (PlexElite 9000, Zeiss) was performed on all eyes. Based on whether an eye received injections or not during follow-up, all eyes were divided into two groups (injection and non-injection group). Vessel density (VD), Vessel skeleton density (VSD) in the superficial, deep, and retina slab, as well as foveal avascular zone (FAZ) size, circularity and perimetry of Angio 6mm×6mm were calculated with algorithms provided by Advanced Research and Innovation (ARI) Network. Change in vascular metrics between baseline and last follow-up were compared between the two groups using t-test or Mann-Whitney U test. Correlation between change in vascular metrics and visual acuity was investigated by Pearson’s correlation test.

Results: A total of 164 eyes from 107 patients were included. The average follow-up time was 34 months. No statistically significant difference in baseline vascular metrics was detected between the injection group (57 eyes) and non-injection (107 eyes) group. The injection group received 12.56 injections during follow-up on average. Among all the parameters, only change in FAZ size during follow-up showed a statistically significant difference between the two groups (0.03（0.01,0.09）vs. 0.02(-0.02,0.06) mm2, P=0.043). Though the injection group had a barely larger FAZ, the difference was not clinically important. No correlation was found between change in vascular metrics and change in visual acuity (P>0.05).

Conclusions: In this retrospective longitudinal study of 164 eyes, repeated intravitreal anti-VEGF injections was associated with no relevant significant changes in OCTA vascular metrics over time.

Candidate: Muhammad Abidi

Poster #: 21

Cost Savings of the FIRST-ROP and TWO-ROP AlgorithmsEliminating Initial Screening Examinations for Retinopathy of Prematurity in Medium- and Low-Risk Cohorts

Muhammad Abidi, Angelica Piccini, Hannah Hwang, Francisco Altamirano, Melissa Yuan, Sandra Hoyek, Celine Chaaya, Nimesh A. Patel

Purpose: Our purpose was to examine the financial impact and savings when retinopathy of prematurity (ROP) screening is optimized to reduce unnecessary exams and healthcare costs. This cost-benefit analysis evaluates savings from risk-stratified screening algorithms with comparable safety to traditional screening.

Methods: The validated TWO-ROP (2023) algorithm proposes a single screening at 36- or 40- weeks postmenstrual age (PMA) for low-risk infants meeting ≤1 of: birth weight <1500 g or gestational age <30 weeks. The proposed FIRST-ROP (2025) algorithm initiates screening at 34 weeks PMA for medium-risk infants (birth weight ≥800 g or gestational age ≥27 weeks). We calculated exam counts and per-patient costs using CPT codes and 2024 federal reimbursement rates. National savings were estimated using the WONDER database annual premature birth rate.

Results: Initial inpatient consultations cost $212.17 ($127.05 facility, $85.12 physician); follow-ups cost $128.68 ($76.62 facility, $52.05 physician). Compared to traditional screening, TWO-ROP 36-week screening involves 1 consult and 1 subsequent exam for only 43% of patients, averaging $267.50 (saving $22.61) per patient. TWO-ROP 40-week screening requires only 1 consult, averaging $212.17 (saving $41.32) per patient. FIRST-ROP 34-week screening eliminates the first or first two exams, respectively saving $90.08 and $154.00 per patient. Projected 10-year national savings were: $9,288,866.30 (TWO ROP 36-week), $16,975,495.60 (TWO-ROP 40-week), and up to $35,613,057 (FIRST-ROP).

Conclusions: This study demonstrates that a risk-stratified approach to ROP screening can save millions of dollars annually, while also maintaining patient safety and outcomes. These benefits are crucial for improving access to care in resource-limited settings and in sustainability of the ROP workforce.

Candidate: Francisco Altamirano

Poster #: 22

Structure-Function Correlation With OCT-Derived Macular Thickness and MAIA Macular Microperimetry in Children With Sickle Cell Disease

Francisco Altamirano, Muhammad Abidi, Hanna De Bruyn, Sandra Hoyek, Celine Chaaya, Ju Hyun Jeon, Osama Sorour, Pablo Altschwager, Anne Fulton, Efren Gonzalez, Nimesh A. Patel

Purpose: To understand the association between macular retinal thickness (OCT) and microperimetry sensitivity (MAIA) in children and young adults with sickle cell disease (SCD).

Methods: Retrospective, cross-sectional study of 63 eyes from 34 pediatric patients with SCD. Microperimetry testing was conducted using a 68-stimulus, 10-degree grid under mesopic conditions (MAIA; CenterVue).

Macular retinal thickness (Heidelberg, OCT) was measured across central, parafoveal, and perifoveal areas. Retinal sensitivity data from microperimetry were included only if the test met the reliability criterion of a fixation loss rate ≤30% and if patients had MAIA, OCT, and BCVA collected on the same date to ensure temporal alignment. We utilized Spearman’s rank correlation and GLMM to assess the relationship between macular thickness and sensitivity.

Results: Sixty-three eyes of 34 patients (61.9% male, 61.9% HbSS, 77.8% Black non-Hispanic) were included. The median age and BCVA at testing were 14.6 years (IQR, 12.2-17.5) and 0 logMAR (IQR, 0-0.1) (Snellen 20/20), respectively. 55 eyes (80%) had ≤30% fixation loss and were considered for analysis. After adjusting for age, BCVA, sickle cell genotype, sex, and race and ethnicity, retinal sensitivity was associated with OCT macular thickness, with a 0.060 decibel decrease in sensitivity for every 1 µm decrease in macular thickness (p=0.009). BCVA was not associated with decreased macular sensitivity (p=0.817) nor OCT thickness (p=0.086).

Conclusions: Children and young adults with SCD exhibited reduced visual function on microperimetry, which correlated with retinal thinning in OCT, despite retaining excellent visual acuity. Early recognition and intervention could mitigate the long-term effects of visual dysfunction in this population.

Candidate: Konstantinos Baroutis

Poster #: 23

Retinal Detachment Risk in a Cohort of von Hippel -Lindau Patients at a Major U.S. Tertiary Care Facility

Konstantinos G. Baroutis, Gustavo Sakuno, Sandra Hoyek, Nimesh A. Patel, Joan W. Miller, Demetrios G. Vavvas

Purpose: To report the retinal findings and retinal detachment (RD) outcomes in a large cohort of von HippelLindau (VHL) patients within a Major U.S. Tertiary Care Facility.

Methods: A retrospective cohort study was conducted between January 2005 and August 2024. We identified patients with VHL from Mass General Brigham's Research Patient Data Repository (7+ million patients) and conducted manual chart review of those VHL patients with ophthalmological examinations. A nested 1:2 matched case-control study compared RD cases with controls. Kaplan-Meier analysis estimated cumulative RCH incidence, and conditional logistic regression identified RD risk factors. Patients without RCH were censored at their last follow-up age.censored at their last follow-up age.

Results: Among 159 VHL patients, 104 had comprehensive eye examinations. RCH cumulative incidence increased with age, reaching 50% by age 31 and 87% by age 70. Eighteen eyes (14 patients, 13% of the cohort) developed RD, predominantly combined rhegmatogenous/tractional (94%), with 29% bilateral involvement. Notably, no patient without an RCH developed an RD in our cohort. All RD cases exhibited proliferative vitreoretinopathy (PVR; Grade C:89%, Grade B:11%). Inter-grader reliability for image review was high (96% agreement, Fleiss κ= 0.78). Only one eye achieved anatomical success after a single surgical procedure. Multiple surgeries (median, 2) achieved reattachment in 41% of eyes (7/17), but visual outcomes remained poor (median logMAR BCVA 2.3, hand motion). A case-control (1:2 ratio) analysis revealed that each one-level increase in the number of prior focal treatments (0 to 1 or 1 to 2+ and above sessions) reduced the risk for RD by nearly 7-fold (OR=0.15, 95% CI 0.03-0.63; p=0.01).

Conclusions: This study emphasizes the high lifelong risk and severity of RCH and associated RD in VHL patients. Surgical treatment of RD is frequently complicated by PVR and yields limited visual recovery, underscoring the challenges of late-stage intervention and the need for proactive surveillance and timely therapeutic intervention. All eyes with RD developed PVR, suggesting a potential crucial role for the pVHL/HIF in PVR development that warrants further investigation.

Candidate: Celine Chaaya

Poster #: 24

Retinal Changes on Imaging as Biomarkers of Systemic Activity in Pediatric Sickle Cell Disease

Celine Chaaya, Sandra Hoyek, Colin Lemire, Omar Halawa, Francisco Altamirano-Lamarque, Efren Gonzalez, Nimesh A. Patel

Purpose: Sickle cell disease (SCD) is an inherited blood disorder that affects millions globally, with a particularly notable impact on the pediatric population. The relationship between retinal and systemic findings is an emerging area of interest, particularly in understanding how changes in retinal vasculature can reflect underlying systemic conditions. While they hold potential for assessing systemic disease burden, they are not widely documented for use as clinical indicators of systemic severity, specifically in children with SCD. Therefore, the current study aims to assess the association of retinal imaging and systemic findings in children with SCD.

Methods: Children with SCD aged 18 years or less who had an ophthalmic examination at a tertiary hospital between January 1998 and August 2022 were included. Systemic findings including number of hospitalizations, hemoglobin (Hgb) levels, and time averaged mean velocity (TAMV) in the right (RMCA) and left (LMCA) middle cerebral artery on transcranial doppler (TCD) results were recorded. Retinal imaging findings included total retinal thickness measured on macular optical coherence tomography (OCT; Spectralis OCT2, Heidelberg Engineering), as well as vessel density (VD) of the superficial (SCP) and deep (DCP) capillary plexus, and foveal avascular zone (FAZ) area measured mm OCT angiography (OCTA) scans. Abnormal TCD velocity was defined as more than 170 cm/s.

Results: Six hundred and six eyes (53% males) were included. When adjusting for race and age, central retinal thickness on OCT was positively correlated with TAMV in RMCA (ρ=0.258, p=0.018), and in LMCA (ρ=0.306, p=0.005), and with hemoglobin level (ρ=0.265, p=0.002). Retinal thinning was associated with a greater number of hospitalizations (ρ=0.19, p=0.016). Similarly, a higher VD in the DCP of the inferior-temporal macula positively correlated with TAMV in RMCA (ρ=0.328, p=0.3) and in LMCA (ρ=0.342, p=0.029). Lower VD in the SCP in the inferior temporal macula correlated with higher number of hospitalizations. The relationship between eye examination results and imaging findings varied according to the specific SCD genotype. Specifically, higher Hgb levels correlated to higher prevalence and severity of retinopathy in HbSC, while it correlated with lower prevalence and severity of retinopathy in HbSS genotypes

Conclusions: OCT and OCTA findings are correlated with progression and severity of systemic disease in children with SCD. Imaging parameters were better correlated with systemic markers than visual acuity. The results suggest the quantitative measures on retinal imaging could be used as a biomarker for systemic disease risk and activity.

Candidate: Chong Chen

Poster #: 25

Longitudinal Evaluation of Microvascular Changes and Imaging

Biomarkers Associated With Visual Prognosis in Retinal Artery Occlusion via Multimodal Quantification

Chong Chen, Kayla Nicole Nodecker, Rajiv M. Sastry, Leiyu Wang, Xinyi Ding, Ying Zhu, Ioanna Ploumi, Selin S. Gumustop, Shivesh Himanshu Shah, Nimesh A. Patel, Leo A. Kim, David M. Wu, Demetrios Vavvas, Deeba Husain, Joan W. Miller, John B. Miller

Purpose: To longitudinally assess changes in macular thickness and microvascular metrics in retinal artery occlusion (RAO) patients compared to controls, and to identify imaging biomarkers associated with visual prognosis.

Methods: Design: Retrospective, longitudinal cohort study. Participants: 56 RAO patients (57 eyes) and 27 controls (30 eyes). Methods: Comprehensive ophthalmic evaluations, including macular OCT and 6 × 6 mm swept-source OCT angiography (SS-OCTA), were performed. Retinal thickness, ischemic area size, distance from the ischemic region to fovea, vessel density (VD), perfusion density (PD), and foveal avascular zone (FAZ) were independently assessed by two graders. The predictive value of clinical and imaging indicators was tested using receiver operating characteristic (ROC) curve analysis. Linear regression correlated imaging biomarkers with visual outcomes. Main Outcome Measures: Longitudinal changes in retinal structure and microvasculature, and their associations with final visual acuity (VA).

Results: Among 57 RAO eyes (28 BRAO, 49.12%) with a median follow-up of 83.0 (35.5, 172.0) weeks, retinal thickness significantly decreased over time (p < 0.05), while the ischemic area expanded from 64.97% to 73.58% (p = 0.002). In 24 RAO eyes with serial SS-OCTA scans, VD in superficial (SCP) and deep capillary plexus (DCP) differed significantly between BRAO and CRAO (p ≤ 0.002). Eyes with a baseline ischemic area ≤ 1/3 of the scan area showed increased VD and PD in both plexuses over time (p < 0.05). FAZ size was significantly larger in CRAO compared to BRAO (p = 0.0001), but remain stable over time (p = 0.341). Better baseline VA, greater ischemic distance to fovea, smaller initial ischemic area, higher VD and PD in SCP, and smaller FAZ were associated with better final VA (all p < 0.05, the areas under ROC curve: 0.80-0.89). Multivariable linear regression identified baseline ischemic area size and FAZ size as independent predictors of final VA (p = 0.003 and 0.008).

Conclusions: This cohort demonstrates the progressive ischemia change in RAO over time. In eyes with ≤1/3 ischemia, partial microvascular reperfusion was observed with increased VD and PD. OCTA provides valuable insight into microvascular alterations and visual prognosis. Ischemic area size and FAZ size are served as critical imaging biomarkers for visual outcomes.

Candidate: Molly Munsell

Poster #: 26

Acquired Myelinated Retinal Nerve Fibers: A Case Series

Mary K. Munsell, Gena Heidary, Melanie A. Kazlas, Linda R. Dagi, Eric D. Gaier, Efren Gonzalez

Purpose: Myelinated retinal nerve fibers (MRNF) are present in 0.57-1% of the population [1]. MRNF are generally benign and present at birth. In two subpopulations of interest, however, MRNF are associated with visually significant pathology. In Straatsma syndrome, patients have axial myopia and amblyopia in the eye with MRNF [2]. These authors have previously presented work showing that MRNF are more extensive in patients with Straatsma syndrome. Acquired MRNF have also been rarely reported, often in association with proven or suspected elevated intracranial pressure. We aimed to explore the demographic and clinical characteristics of these two populations of interest.

Methods: Charts were retrospectively reviewed for all patients with MRNF examined at one tertiary center between 2010 and 2025. Straatsma syndrome was defined as spherical equivalent (SE) refractive error less than or equal to -3.0 D and amblyopia with Snellen best corrected visual acuity (BCVA) <20/60 in the eye with MRNF. Acquired MRNF were identified based on documented fundus exams newly noting MRNF after previous documented dilated exams without this finding. Fundus photography, optical coherence tomography, and visual field testing were performed for a subset of patients.

Results: A total of 182 patients with MRNF were identified, of whom 20 met criteria for Straatsma syndrome and 18 had acquired MRNF. Fundus photos documenting optic nerve appearance before and after MRNF onset were available for 7 patients. Of patients with acquired MRNF, 4 had craniosynostosis, 4 had intracranial tumors, 3 had neurofibromatosis, 3 had Chiari-Arnold malformation, and 1 had idiopathic intracranial hypertension. However, 5 patients had no clear medical history to suggest elevated intracranial pressure. No patients with acquired MRNF had associated high myopia and amblyopia. MRNF were bilateral in 6 (33%) of patients with acquired MRNF, compared to 45 (28.5%) of all patients with MRNF.

Conclusions: The pathophysiology of MRNF is incompletely understood. Our large case series supports a temporal role for increased intracranial pressure in many cases acquired after the perinatal period of ophthalmic development. This supports theories that the extension of myelination into the retina is due to mechanical compromise of the lamina cribrosa [1]. However, the presence of otherwise healthy patients in the acquired MRNF cohort could also suggest multiple etiologies of MRNF. The lack of overlap between the Straatsma syndrome and acquired MRNF populations may suggest that the myopia associated with Straatsma syndrome rather than myelinated fibers themselves are visually significant and amblyogenic.

Candidate: Jia Jia Zhang

Poster #: 27

Evaluation of Systemic Diagnostics,

Management Strategies and Outcomes in Admitted vs. Non-Admitted Patients With Central Retinal Artery Occlusion

Jia Jia Zhang, Sandra Alhoyek, Saghar Bagheri, Melissa Yuan, Ryan S. Meshkin, Celine Chaaya, John B. Miller, David N. Zacks, Joan W. Miller, Demitrios Vavvas, Nimesh A. Patel

Purpose: There is limited research that examines how systemic management, follow-up, and outcomes differ between admitted vs. non-admitted patients with central retinal artery occlusion (CRAO).

Methods: CRAO patients were identified with ICD Code H34.1 and manual chart review performed for demographics, diagnostic testing, findings, time to presentation, admission, and initial testing, treatments, PCP follow-up, subsequent admissions within 2 years, and mortality. Statistical analyses included chi-squared, Wilcoxon rank-sum, and T-tests.

Results: The study included 107 admitted and 38 non-admitted CRAO patients. Both groups were similar in presentation age, sex, and race, though more Hispanic patients were not admitted (p=0.013). Median hours to presentation was comparable 8.5 vs 10.35 (p=0.4). Non-admitted patients often had no initial imaging (p<0.001); however, the proportion of positive findings (stenosis, stroke, or thrombus) for each modality was similar (MRI p=0.624, CT p=0.059, carotid US p=0.16, echo p=0.611). Admitted patients underwent more blood tests (p<0.001) and procedures, particularly ipsilateral carotid endarterectomy (11.5% vs 0 %, p=0.035). Outpatient testing was higher in non-admitted patients (39.5% vs. 19.6%, p<0.001). However, readmissions rate within 2 years (p=0.160), PCP follow-up (p=0.078), and mortality rates (20.6% vs. 13.2%, p = 0.863) were comparable.

Conclusions: These results suggest that intensive inpatient management may not improve readmission rates and all-cause mortality in patients with Central Retinal Artery Occlusion. Outpatient medical management with timely imaging and follow-up could be a comparable option.

Candidate: In Young Chung

Poster #: 28

Bandage Contact Lens Retention Following Boston Keratoprosthesis

Type I Implantation

In Young Chung, Michael Bednar, Eleftherios Paschalis, Thomas H. Dohlman

Purpose: Full-time bandage contact lens (BCL) wear is recommended following Boston Keratoprosthesis Type I (KPro I) implantation to protect the ocular surface, maintain hydration, and promote device retention. However, some patients fail to retain BCLs due to various ocular surface or anatomical challenges. This study aims to characterize BCL retention outcomes and identify potential risk factors associated with BCL loss in patients who underwent KPro I implantation.

Methods: This study included 35 eyes from 35 randomly selected patients who underwent Boston Keratoprosthesis Type I (KPro I) implantation between 2012 and 2021. Clinical data were collected through retrospective chart review, including patient demographics, ocular history, preoperative diagnosis, BCL type, and retention status at the final follow-up. The primary outcomes were BCL retention, duration of wear, BCL dimensions and associated ocular surface findings. Descriptive statistics were used to summarize the data.

Results: Of the 35 eyes included, the mean patient age was 63.0 ± 18.9 years, and 57.1% (n=20) were male. The average follow-up duration from the time of surgery to the last visit was 5.7 ± 4.6 years. At the final followup, 26 out of 35 eyes (74.29%) retained a BCL. At last follow-up, the mean duration of retention for that BCL was 4.97 ± 3.12 months. Among the 26 eyes that retained a BCL at final follow-up, the most commonly used BCL type was the Kontur lens (19/26; 73.08%). The most commonly used Kontur lens dimension was 16 mm diameter/8.9 mm base curve (12/19, 63.16%), followed by 18 mm/9.0 mm (3/19; 15.79%), 16 mm /8.6 mm (2/19; 10.53%), 15 mm /8.6 mm (1/19; 5.26%), 14 mm /8.9 mm (1/19; 5.26%) and one case with unknown specifications (1/19; 5.26%). The most lenses were plano (21/26; 80.77%). Among patients with documented refractive power (3/26; 11.54%), the prescriptions were -16D, +3D, +10D. The most common indication for KPro implantation was corneal edema (5/35; 14.29%). Of the 7 eyes (20%) that failed to retain a BCL, the contributing factors included: 1) Hypotony causing globe contour changes; 2) excessive epiphora; 3) gelatinous drop-like dystrophy of the host cornea; 4) recurrent symblepharon; 5) inferior symblepharon, and fornieal shortening; 6-7) two cases of unknown etiology.

Conclusions: Approximately three-quarters of KPro I eyes were able to retain a bandage contact lens at final follow-up. BCL loss was associated with hypotony, excessive epiphora, host corneal surface irregularities, symblepharon, and forniceal shortening. Identifying and mitigating these risk factors may help guide efforts to improve contact lens retention and long-term outcomes in KPro patients.

Candidate: Jonathan Deck

Poster #: 29

A Case of Keratomalacia in Severe Vitamin A Deficiency

Jonathan Deck, Athena Cohen, Joseph Raevis, Jae Young You

Purpose: Vitamin A is a lipid-soluble vitamin essential for vision, playing a critical role in the function and health of the retina and cornea. While vitamin A deficiency is a common global health issue, it is relatively rare in the United States, typically occurring in the context of malnutrition or malabsorption syndromes. Xerophthalmia encompasses the spectrum of ocular manifestations associated with vitamin A deficiency, ranging from night blindness to keratomalacia. In this report, we describe a severe case of xerophthalmia manifesting as keratomalacia in a 42-year-old woman with short bowel syndrome.

Methods: A 42-year-old woman with history of Graves’ disease and alcohol use disorder presented to the emergency department with five weeks of progressive right eye (OD) pain, redness, and decreased vision. On initial examination, uncorrected visual acuity was hand motion OD and 20/30 in the left eye (OS). Intraocular pressure was normal in both eyes (OU). Slit-lamp examination OD revealed highly inflamed conjunctiva with diffuse injection, as well as exceptional corneal ectasia with 360-degree thinning at the limbus, a small superotemporal descemetocele, and diffuse stromal haze. The anterior chamber appeared shallow with the iris diffusely apposed to the corneal endothelial surface, but the Seidel sign was negative. There was no posterior view. B-scan ultrasonography demonstrated no vitritis and an attached retina OU. OD examination was remarkable for peripheral superior and inferior corneal pannus with superonasal corneal neovascularization. Further questioning of the patient revealed pertinent past surgical history including a surgical complication from the repair of an enterocutaneous fistula, which required multiple revisions and ultimately resulted in short bowel syndrome. The patient had been on total parenteral nutrition (TPN) until two years ago, when she was transitioned to enteral nutrition due to recurrent line infections.

Results: Subsequent evaluation revealed an undetectable vitamin A level (<5 mcg/dL) along with multiple other nutritional deficiencies. The combination of her slit lamp examination findings, undetectable vitamin A level, and short bowel syndrome led to the ultimate ophthalmic diagnosis of keratomalacia in the setting of extreme vitamin A deficiency. She was admitted to the hospital for TPN and aggressive vitamin A supplementation was initiated to correct her deficiency and prevent further ocular deterioration. Ultimately, this patient’s care was transferred to a drug rehabilitation facility due to her history of alcohol and substance use. The details of the patient’s substance use history could not be obtained. Long-term implications for patients with vitamin A deficiency, particularly those with underlying gastrointestinal issues, necessitate a comprehensive treatment approach. Managing the underlying conditions contributing to malabsorption, as seen in this case with short bowel syndrome, is crucial for preventing recurrence of deficiency-related complications.

Conclusions: In the context of this case, the patient’s history of Graves’ disease, short bowel syndrome, and chronic alcohol use provided a unique set of circumstances leading to the advanced and unusual presentation of vitamin A deficiency. Our patient presented with the most severe features of xerophthalmia including keratomalacia, corneal ulcers and a descemetocele OD.

Candidate: Ju-Yeun Lee

Poster #: 30

Factors Associated With Visual Axis Opacification Following Pediatric Cataract Surgery in the IRIS® Registry (Intelligent Research in Sight)

Ju-Yeun Lee, Ju Hyun Jeon, Francisco Altamirano, Nadine AlMuasher, Tobias Elze, Joan W. Miller, Alice C. Lorch, Deborah K. VanderVeen, Isdin Oke, IRIS® Registry Analytic Center Consortium

Purpose: To determine the probability of visual axis opacification (VAO) following pediatric cataract surgery and to identify associated risk factors.

Methods: This retrospective cohort study included all children who underwent cataract surgery between 2013 and 2020 in the IRIS® Registry (Intelligent Research in Sight). Data collected included age, sex, race and ethnicity, insurance type, geographic region, cataract laterality, history of trauma, uveitis, retinopathy of prematurity, intraocular lens placement, and anterior vitrectomy. The two-year cumulative probability of VAO was determined using the Kaplan-Meier estimator. Multivariable Cox proportional hazard regression models were used to assess factors associated with VAO diagnosis and treatment. Main Outcome Measures were cumulative probability of VAO diagnosis and treatment within two years of pediatric cataract surgery and hazard ratios (HR) of risk factors.

Results: A total of 7,075 children (median [Interquartile range], 11 [5-16] years, 53% male) were included. The cumulative probability of VAO diagnosis within two years was 31.3% (95% CI, 29.4-33.1) and VAO treatment within two years was 23.3% (95% CI, 21.6-25.0). Children undergoing surgery at an older age (12 to <19 years vs. 0 to <3 years; HR=2.13, 95% CI:1.69-2.69), unilaterally (HR=1.29, 95% CI 1.16-1.43), with intraocular lens placement (HR 4.14, 95% CI:3.17-5.41), and without anterior vitrectomy (HR 2.15, 95% CI:1.58-2.92) were at increased risk for developing VAO. Children with a history of ocular trauma (HR=1.67, 95% CI 1.32-2.13), retinopathy of prematurity (HR=1.34, 95% CI:1.15-1.55), anterior uveitis (HR=1.81, 95% CI:1.30-2.51) and posterior uveitis (HR=1.72, 95% CI:1.03-2.88) were also at increased risk. Similar associations were identified for VAO treatment.

Conclusions: Identifying potential risk factors for VAO is essential to prevent further vision loss from amblyopia in pediatric patients. Results from this study support the need for close postoperative monitoring of children undergoing cataract surgery, especially those with a history of trauma or uveitis to enable early diagnosis and treatment of VAO.

Candidate: Angelica Piccini

Poster #: 31

Infectious Keratitis in the Pediatric Population at a Tertiary Referral Center.

Angelica Piccini, Dennis Akrobetu, Asmaa A. Zidan, Tatiana Hathaway, Thomas H. Dohlman

Purpose: Infectious keratitis (IK) is a common ophthalmic condition that can cause permanent vision loss and has been studied extensively in adults; however, little is known about the risk factors and impact of the condition in the pediatric population (patients < 18 years) within the United States. Investigating the etiologies and visual outcomes in this population is critical given the risk of amblyopia and potential long-term visual deficits. This study investigates the demographics, causes, visual burden, treatments, and culture results of pediatric patients examined through the Massachusetts Eye and Ear cornea service and diagnosed with IK (meeting culture criteria) between 2003 to 2023.

Methods: Patients meeting inclusion criteria were found by accessing the Research Patient Data Registry and identifying patients who had corneal microbiology cultures collected and at least one of the following ICD-10 codes: H16, H16.0, H16.00, H16.002, H16.003, H16.009, H16.01, H16.011, H16.012, H16.013, H16.02, H16.021, H16.022, H16.023, H16.029, H16.03, H16.031, H16.032, H16.033, H16.039, H16.07, H16.1, H16.12, H16.20, H16.21, H16.22, H16.23, H16.25, H16.26, H19.1, 53.29, B02.3, B02.20, B02.33, B02.39, 370.55, 370.35, H16.23, 370.52, 370.5. Data on age, gender, underlying etiology, microbial culture results, treatment strategies, and best corrected visual acuity (BCVA) before, at, and three months after presentation were collected from included patients.

Results: A total of 31 patients were included. 29.03% were male; the median age was 11 years with an interquartile range (IQR) of 9 and 15 years. For patients with Snellen data, median best corrected visual acuity (BCVA) was 1.3010 before diagnosis (IQR 0.1365-1.3979); 0.3495 at diagnosis (IQR 0.1761-1.0326); and 0.6505 at three months (IQR 0.2698-0.7959). Streptococcus was the most common microbe isolated on culture. All patients were treated with ophthalmic fortified drops. Less frequently, some received oral antibiotics (n=4, 12.90%) or oral antivirals (n=5, 16.13%). One patient received intravenous antivirals (n=1, 3.23%). Surgical intervention was employed in 10 (32.26%) patients.

Conclusions: There is a diverse array of underlying etiologies for IK in the pediatric population. In this pediatric cohort at a tertiary referral center, Streptococcus was the most frequently identified microbe for cultures which resulted in a specific organism. The median BCVA significantly declined at presentation compared to baseline. Although there was improvement in median recorded BCVA by three months, it did not return to pre-diagnosis levels.

Candidate: John Fallon

Poster #: 32

Ocular Surface Mast Cells Contribute to Injury-induced Neuroinflammation

John M. Fallon, WonKyung Cho, Meghanashree M. Shreenivas, Kanika Arora, Lei Xi, Vinay K. Pulimamidi, Olufemi S. Folorunso, Nishant R. Sinha, Sunil

K. Chauhan

Purpose: Mast cells are sentinel immune cells that are essential for the innate immune response. While their activation following ocular surface injury is known to induce inflammatory tissue damage, their role in corneal nerve damage remains unclear. To address this, this study examined mast cell infiltration and activation in proximity to corneal nerves after injury and assessed their contribution to trigeminal neurite degeneration.

Methods: Corneal injury was induced in female C57BL/6 mice through mechanical removal of the corneal epithelium and anterior stroma. Corneas were harvested 6h following injury and labeled for mast cells (avidin+) and corneal nerves (Beta-tubulin III+) for epifluorescence analysis. Trigeminal ganglia were harvested postinjury, and lysates were analyzed for mast cell activation (tryptase) and leukocyte frequency (CD45+) through ELISA and flow cytometry, respectively. To study the interaction between mast cells and corneal nerves, trigeminal ganglia were co-cultured with bone-marrow derived mast cells for 24h ± mast cell inhibitor cromolyn (100 μM). Trigeminal ganglia were then assessed for neurite degeneration by quantifying levels of Substance P and neurite length (Beta-tubulin III+; quantified by ImageJ).

Results: Following injury, mast cells infiltrated the cornea and were observed adjacent to damaged nerves. Corneal injury was associated with increased mast cell activation (tryptase; p=0.003) and leukocyte frequencies (CD45+; p=0.01) in the trigeminal ganglia compared to naïve. In vitro, co-culture of mast cells with trigeminal ganglia resulted in a 50% decrease in neurite length and a 4-fold increase (p<0.0001) in the expression of substance P, indicative of neurite degeneration. Mast cell inhibition with cromolyn in the mast cell-trigeminal ganglia co-culture suppressed mast cell-induced neurite degeneration (p<0.0001).

Conclusions: Mast cells aggregate around nerves in the injured cornea, contributing to nerve damage at both the ocular surface and within the trigeminal ganglia.

Candidate: Seokjoo Lee

Poster #: 33

Myeloid-Derived Suppressor Cells

Enhance

Wound Healing in a Murine Model of Corneal Injury

Seokjoo Lee, Shweta Sandhu, Neda Heydarian, Asmaa A. Zidan, Vinay K. Pulimamidi, Lei Xi, Yihe Chen, Jia Yin, Sunil K. Chauhan, Reza Dana

Purpose: Myeloid-derived suppressor cells (MDSC) are innate immunoregulatory cells that are early responders to tissue injury, playing a key role in tissue repair and wound healing. However, their specific function in corneal injury, particularly in enhancing wound healing, has not been thoroughly investigated. This study aims to evaluate the role of MDSC in promoting corneal wound healing.

Methods: CD11b+Gr-1+ MDSC and CD11b+Gr-1- cell control were isolated from cultured BALB/c bone marrow using magnetic-activated cell sorting. For the in vitro scratch assay, 2.5 × 10⁵ human corneal epithelial cells (hCEC) were co-cultured with either 2.5 × 10⁵ MDSC or cell control. The defect area was assessed at 18and 24-hours using ImageJ. Corneal injury was induced in BALB/c mice by removing the epithelium using a hand-held Algerbrush-II. MDSC or cell control (50,000 cells in 50 µL saline) were injected subconjunctivally. Corneal epithelial healing was assessed through fluorescein staining and slit lamp photography at 6, 22, and 28 hours, with the images analyzed using ImageJ. At day 3, corneas were harvested, and flow cytometry was used to determine CD11b+Ly6G+ neutrophil percentages.

Results: The scratch assay using hCEC revealed that MDSC-treated cells exhibited a significantly smaller defect area at 18 hours compared to those treated with the cell control (p = 0.01) or saline (p = 0.02), but no significant difference was observed between the groups at 24 hours. In the in vivo corneal epithelial injury model, the MDSC-treated groups demonstrated significantly accelerated healing compared to the cell control (p = 0.0003) and saline-treated groups (p = 0.016) at 6 hours. By 22 hours, the wound healing area among the groups were nearly identical, with complete healing observed in all groups by 28 hours. Flow cytometry analysis showed significantly lower neutrophil percentages in both the MDSC-treated group (0.87 ± 0.12%, p = 0.0004) and the cell control group (1.27 ± 0.17%, p = 0.0032) compared to the saline-treated group (2.07 ± 0.12%). Although the MDSC-treated group had a modestly lower mean neutrophil percentage than the cell control, this difference was not statistically significant (p = 0.067).

Conclusions: Our findings demonstrate that MDSC enhance the rate of tissue repair by minimizing epithelial defect areas, reducing neutrophil infiltration, and effectively supporting corneal wound healing.

Candidate: Meghanashree Malavvar Shreenivas

Poster #: 34

Mast Cells Amplify Neutrophil Activation and Secretion of TissueDamaging Factors

Meghanashree M. Shreenivas, John M. Fallon, Kanika Arora, Vinay K. Pulimamidi, Lei Xi, Olufemi S. Folorunso, Nishant R. Sinha, Sunil K.

Chauhan

Purpose: Neutrophils play a vital role in ocular immunity by exerting their pro-inflammatory functions necessary for the control and clearance of ocular infections. However, in response to injury, prolonged neutrophil activation and excessive secretion of their effector molecules result in tissue damage. Previous studies from our lab have demonstrated that mast cells initiate neutrophil recruitment to the ocular surface following injury. Here, we investigated the role of mast cells in promoting the activation of neutrophils and their secretion of tissue-damaging effectors.

Methods: Bone marrow cells were harvested from femurs and tibias of C57BL/6 mice and cultured for 3-4 weeks in the presence of IL-3 (10 ng/ml) and stem cell factor (50 ng/ml) to generate bone marrow derived mast cells (BMMCs). BMMCs were then stimulated with either PMA (50 ng/ml), IL-33 (100 ng/ml) or compound 48/80 (1 µM). The supernatant was collected to evaluate the colony stimulating factors by enzyme linked immunosorbent assay (ELISA). Subsequently, neutrophils were sorted from the bone marrow cells of mice using a neutrophil isolation kit (>80% purity). Neutrophils were co-cultured with activated BMMC supernatant for 3 hours. Thereafter, neutrophils in the co-culture were analyzed to evaluate the expression of activation associated markers (CD11b, Ly6G, CD63 and CD62L) using flow cytometry. Supernatants of the co-cultures were collected to assess the quantity of myeloperoxidase (MPO) and neutrophil elastase (NE) using ELISA.

Results: Mast cells stimulated with IL-33 secreted ~2000 pg of Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) at 24h. Neutrophils co-cultured with activated mast cell supernatant showed a significant increase in the expression (MFI) of maturation markers CD11b (p<0.0001), Ly6G (p<0.0001), CD63 (p<0.0001) as compared to neutrophils cultured with resting mast cell supernatant. Further, a significant reduction in the expression of cell adhesion molecule CD62L was observed (p<0.0001). The expression of CD62L negatively corelates with neutrophil activation. Additionally, increase in MPO and NE, enzymes secreted by neutrophils which are involved in tissue damage, was observed in the supernatant of neutrophils co-cultured with activated mast cell supernatant. GM-CSF neutralization in the activated mast cell supernatant lead to significant decrease in the activation markers expressed by neutrophils as well as secretion of tissue damaging factors like MPO and NE.

Conclusions: Our data demonstrate that mast cells promote neutrophil activation and secretion of tissuedamaging effectors in a GM-CSF dependent manner. Future experiments are aimed at understanding the molecular mechanisms of mast cell-neutrophil interactions.

Candidate: Jyoti Sharma

Poster #: 35

Pre-clinical Outcomes of a Novel Pan Ocular Therapy for Ocular Injuries

Jyoti Sharma, Bin Yan, Maria Emfietzoglou, Chengxin Zhou, Eleftherios I. Paschalis.

Purpose: Severe injuries to the eye, as encountered in the military and general public, can lead to corneal and retinal damage and subsequent blindness. We developed a thermosensitive drug delivery system (DDS) for sustained delivery of TNF-α and VEGF inhibitors to the eye for 3 months and showed marked pan-ocular protection when given immediately after injury (Zhou et al., 2023). To transition this therapy to military reality, we evaluate the protective effect when given 1 or 3 days after the injury, assuming delays in patient evacuation to ROC level 3 or 4

Methods: New Zealand white rabbits (n=18) received severe corneal alkali burn with 2N NaOH, applied to the corneal surface for 20 sec using an 8 mm in diameter filter paper. Eyes received copious irrigation for 15 minutes with saline solution. One or 3 days later, the eyes were injected subconjunctivally with DDS (PLGAPEG-PLGA triblock polymer) containing 0.7mg adalimumab and 1.3mg aflibercept. Control animals received DDS with 2mg of IgG. Animals were followed biweekly for 3 months with clinical photography, in vivo optical coherent tomography, fluorescein staining, and intraocular pressure (IOP) assessment with cannulation. At the end of the study, eyes were enucleated and analyzed histologically with hematoxylin & eosin and pPhenylenediamine staining, for retinal and optic nerve status, respectively.

Results: Administration of the anti-TNF-α/anti-VEGF DDS, 1 or 3 days after the injury effectively reduced corneal neovascularization (p=<0.0001) and accelerated corneal wound healing (p=<0.0001), as compared to IgG therapy. IgG DDS lead to 42.7% increase in IOP, as compared to baseline (11.8±1.8 mmHg) measurements, whereas anti-TNF-α/anti-VEGF DDS caused minimal change in the IOP, with only 4.3% or 2.1% increase when the drug was administered 1 or 3 days after the injury, respectively. In addition, anti-TNFα/anti-VEGF DDS prevented retinal ganglion cell (p=0.03) and optic nerve axon (p=0.02) loss when administered 1 day after the injury. This protective effect was abolished in animals treated 3 days after the injury (p=0.3)

Conclusions: Delayed (3 days) administration of the thermosensitive anti-TNF-α/anti-VEGF DDS prevents corneal neovascularization and expedites wound healing following a corneal alkali burn. Notably, when administered 1 day after the injury, it also provides retinal and optic nerve protection. This proposed therapy is practical for military use and has the potential to significantly enhance the treatment of ocular alkali burns and possibly other types of injuries

Candidate: Bhupender Verma

Poster #: 36

Supersaturated Oxygen Emulsion as a Therapeutic Strategy for Nitrogen Mustard-induced

Intraocular Inflammation and Retinopathy

Bhupender Verma, Asmaa A. Zidan, Sheyda Najafi, Hennaav K. Dhillon, Jia Yin

Purpose: Mustard agents like nitrogen mustard (NM) are vesicating chemicals classified as chemical warfare agents that can lead to ocular pain and vision impairment, yet no effective treatment currently exists. Although ocular surface damage after NM exposure is extensively studied, its impact on intraocular inflammation, retina damage, and underlying mechanisms remain elusive. We previously demonstrated that perfluorodecalin-based supersaturated oxygen emulsion (SSOE) is safe and effective in treating corneal burns. In this study, we investigate the efficacy of topical SSOE in mitigating intraocular inflammation and retinopathy after NM exposure.

Methods: Adult BALB/c mice were exposed to NM by topical application of 3μl of 25mM NM solution on the ocular surface for 5 minutes. Topical SSOE or vehicle were applied once daily for 14 days. To assess intraocular inflammation, infiltrating CD45+ cells in the iris were quantified using flow cytometry and TNF-α levels in the iris and aqueous humor (AH) were measured using ELISA. Neurosensory retina thickness from hematoxylin and eosin-stained tissue sections was quantified using imageJ. Immunostaining for glial fibrillary acidic protein (GFAP) and glutamine synthetase (GS) in the retina was performed to assess gliosis.

Results: NM exposure significantly upregulated infiltrating CD45+ lymphocytes in the iris at day 1 and 14 after the injury. Topical SSOE application significantly reduced the frequency of infiltrating CD45+ lymphocytes in the iris at day 1 as well as at day 14. NM led to significant upregulation in TNF-α levels in the iris and AH, which was significantly reduced by SSOE treatment. At Day 28, untreated mice exposed to NM exhibited significant retinal thinning, decreased GS expression, and upregulated GFAP expression. SSOE treatment significantly reduced the retinal thinning and maintained homeostatic GS and GFAP expression in the retina.

Conclusions: NM exposure to the ocular surface leads to intraocular inflammation and retina damage. Topical SSOE application dampens pro-inflammatory response in the eye and mitigates reactive gliosis and retinal damage, underscoring its therapeutic potential in mitigating mustard agent-induced eye injury.

Candidate: Camille Andre

Poster #: 37

Host and Microbial Virulence Factors That Drive Sight-threatening Methicillin-resistant

S. aureus Infections

Camille André, Adam Pickrum, Victor Torres, Paulo J.M. Bispo, Michael S. Gilmore

Purpose: S. aureus is a major cause of eye infections. Methicillin-resistant S. aureus (MRSA) is commonly associated with multidrug-resistant (MDR) infections, resulting in treatment failure and blindness. By performing a large in-depth genomic characterization of ocular MRSA isolates (n=247), we found that the major epidemic clones clones, CC5/USA100 and CC8/USA300, exhibited different ocular tissue tropism. CC5 strains were being more trophic to the wet epithelial surface while CC8 MRSA strains were more prone to cause skin and soft tissue infections of the ocular adnexa. Despite this distinct ocular biogeography, it is not understood what host and microbial factors drive epidemic MRSA CC5 and CC8 ocular tissue tropism?

Methods: To assess the emergence of high-risk MDR clones of MRSA causing eye infections we have sequenced the genomes of all MRSA isolates recovered from patients presenting with eye infections the past 8 years at MEE. Virulence factor database software was used to identify virulence genes and mutations that contribute to the pathogenicity of the bacteria. We also used bioinformatic pipeline to identify strain-specific virulence factors. Because neutrophil phagocytosis is the first and key element of host immune defense of the ocular surface, we assessed cytotoxicity toward neutrophils and resistance to phagocytosis of the two major epidemic MRSA lineages causing eye infections.

Results: Most of these patients were female (59.4%). Age at presentation ranged from 1 to 103 years (median 53 years old). 90% of MRSA CC5 isolates were MDR (resistance to ≥3 antimicrobial classes). In line with these observations, we found that the resistome of these CC5 strains was comprised of multiple acquired antimicrobial resistance genes (AMR) that confer resistance to various clinically important antibiotics. We discovered strain-specific virulence factors. Three superantigen toxins (sed, sej and ser) were found to be significantly enriched among CC5 ocular isolates, whereas Panto-Valentine Leukocidin (PVL) was enriched in CC8. Moreover, we saw a difference in the distribution of PVL among CC8 isolates with 91.7% of MRSA CC8 causing periocular infections being PVL+ whereas only 21.4% of MRSA CC8 causing keratitis were PVL+. We also identified specific mutations in the alpha-toxin gene that may contribute to the eye pathogenesis. We found that MRSA CC8 isolates were more cytotoxic to human polymorphonuclear leukocytes (hPMNs) compared to MRSA CC5 isolates. However, this did not come at the cost of the ability to survive hPMN attack. Though less cytotoxic, CC5 isolates were significantly more fit to survive in hPMNs.

Conclusions: Epidemic MRSA isolates causing eye infections show a distinct virulence factors repertoire and host response, driving patterns of infectious diseases that we observed in ophthalmology.

Candidate: Kanika Arora

Poster #: 38

Mesenchymal Stem Cells Suppress the Proliferation of T cells by Inducing Anti-proliferative Tob1 Expression

Kanika Arora, Meghanashree M. Shreenivas, Vinay K. Pulimamidi, Nishant R. Sinha, John M. Fallon, Olufemi S. Folorunso, Lei Xi, Sunil K. Chauhan

Purpose: Mesenchymal stem cells (MSCs) have the unique capacity to regulate inflammatory process. Our lab has previously shown that MSCs suppress the generation of alloreactive Th1 cells. However, the mechanisms underlying this regulation remain unknown. In this study, we investigated the cellular and molecular mechanisms by which MSCs suppress T cell proliferation.

Methods: Human bone marrow-derived MSCs (STEMCELL Technologies, Canada) were expanded using MesenCult™-ACF Plus Culture Kit (STEMCELL Technologies) using the plastic adherence method. MSCs were characterized phenotypically as CD45-CD34-CD90+CD73+ by flow cytometry. CD4+ T cells were isolated from human PBMCs using a CD4 isolation kit. Sorted naïve CD4 T cells were stimulated with CD3/CD28 beads and cocultured with and without MSC. Expression of Tob 1 was quantified using real-time qPCR. Sorted naïve T cells were labelled with CFSE and then co-cultured with MSC or Tob 1 agonist (25µM). Proliferation was measured at 72 hours using flow cytometry. All invitro experiments were performed in quadruplets and repeated three times.

Results: Flow cytometry data of human MSCs showed negative expression of hematopoietic markers CD45 and CD34 and positive expression of stromal cell markers CD90 and CD73. Stimulated CD4+ T cells showed a significant 78% reduction in the expression of Tob1 mRNA as compared to naïve CD4+ T cells. However, Tob1 expression in the MSCs co-cultured with activated CD4+ T cells showed a substantial 48% increase as compared to activated CD4+ T cells. Proliferation of CD4+ T cells stimulated with CD3/CD28, was found to be lower when co-cultured with MSCs. Administration of Tob1 agonist showed a similar reduction in proliferation as was observed in CD4+T cell and MSC co-culture.

Conclusions: Our data suggests that MSC prevent downregulation of Tob1 in naive T cells upon their TCRmediated activation and inhibit proliferation. Future studies are aimed at understanding the molecular mechanisms by which MSCs induce Tob 1 expression in T cells.

Candidate: Yeganeh Farsi

Poster #: 39

Pathogenic Function of T helper 17 Cells on Corneal Epithelial Cells in Dry Eye Disease

Yeganeh Farsi, Seokjoo Lee , Neda Heydarian, Meghanashree M. Shreenivas, Rohan B. Singh, Yihe Chen, Reza Dana

Purpose: To investigate the differential pathogenic capacity of effector and memory Th17 cells on corneal epitheliopathy.

Methods: Acute DED was induced in 6-8-week-old female IL-17A-GFP reporter mice by subjecting them to desiccating stress for 14 days. Chronic DED was induced by transferring the mice to a standard vivarium for an additional 14 days. Post-induction, CD4+T cells were isolated from the draining lymph nodes of acute and chronic DED mice using magnetic-activated cell sorting. Subsequently, Th17 cells (profiled as CD4+L-17A+) were isolated by fluorescence-activated cell sorting. Naive CD4+T (Th0) cells from normal mice served as controls. The purified Th17 or Th0 cells were then cocultured with normal CEpCs (1:3) for 24 hours. Following co-culture, CEpCs were assessed for early (Annexin V+ 7-AAD-) and late apoptosis/necrosis (Annexin V+/- 7AAD+) via flow cytometry. The expression levels of Zonula Occludens-1 (ZO-1) and inflammatory cytokines (TNF-a and IL-6) in CEpCs were quantified by RT-qPCR. Also, the expression of Zo-1 at protein level was investigated by Immunocytochemistry.

Results: CEpCs cocultured with acute Th17 cells showed a significantly higher frequencies of early apoptotic cells compared to those cocultured with Th0 cells (58.5+0.7%vs.44.8+0.9%,p=0.0009) or CEpCs alone (58.5+0.7%vs.47.4+0.4%,p=0.0016).However, the frequencies of early apoptotic cells on coculturing CEpCs with chronic Th17 cells were comparable to that observed in the coculture with Th0 or CEpCs alone. While effector Th17 cells (derived from acute DED mice) induced higher TNFα expression in co-cultured CEpCs, memory Th17 cells (derived from chronic DED mice) significantly increase both TNFα and IL-6 expression in CEpCs. Zonulin-1 (ZO-1) expressed significantly lower in CEpCs cocultured with effector Th17 cells compared to other groups.

Conclusions: Th17 cells cocultured with CEpCs lead to increased apoptosis, higher expression levels of proinflammatory cytokines, and decreased expression of ZO-1. Th17-mediated pathogenic effects on corneal epithelial cells are more pronounced in eTh17 than in mTh17 cells.

Candidate: Olufemi Folorunso

Poster #: 40

Membrane-type II Metalloproteinase (MMP15) Alters Corneal Collagen Architecture

Olufemi S. Folorunso, Vinay K. Pulimamidi, Lei Xi, Nishant R. Sinha, Meghanashree M. Shreenivas, Kanika Arora, John M. Fallon, Sunil K. Chauhan

Purpose: Corneal collagen homeostasis is essential in maintaining the stromal extracellular matrix. Corneal fibroblasts regulate stromal collagen turnover through matrix metalloproteinases (MMPs) remodeling activities. The functions of membrane-type metalloproteinases (MT-MMPs) are not well defined in corneal pathophysiology. In this study, we analyzed the differential expression of MMPs in normal and pathological corneas (keratoconus) using publicly available RNA-seq from the NCBI GEO2R database to identify a highly significant expressed MMP associated with corneal pathology. The function of the selected MMP was further validated in corneal cells and an animal model of corneal injury.

Methods: We explored the RNA-seq bulk datasets containing data from pathological (keratoconus) and normal corneas. To corroborate our in-silico findings, corneal fibroblasts and epithelial cells were evaluated for the expression of selected MMP in homeostasis and inflammatory milieu. Also, an in vitro assay involving ECM mixed with collagen type I was standardized to evaluate whether corneal cells in homeostatic and inflammatory conditions degrade collagen type I into soluble collagen fragments that were quantified using an ELISA assay. Ultimately, we determined if the selected MMP could maintain corneal stroma architecture under pathological conditions.

Results: MMP15 was differentially expressed at a higher level in the cornea with stromal pathology. The expression was significantly higher in the keratoconus cornea stromal cells than in the normal, suggesting the role of MMP15 in stromal ECM homeostasis. Unlike the corneal epithelia, IL-1β-stimulated corneal fibroblasts significantly expressed MMP15 compared to homeostatic cells, predicting that MMP15 expression is inflammation-dependent. MMP15 activities correlated with collagen degradation in vitro under inflammatory conditions. Blocking MMP15 in the corneal stromal injury prevented stromal abnormality while promoting wound healing by downregulating the expression of other collagen-degrading MMPs

Conclusions: Our data demonstrate that corneas with common stromal pathologies (e.g., keratoconus) express a high level of membrane type II metalloproteinase. The significant expression of MMP15 is primarily associated with stromal cells (i.e., fibroblasts) under inflammatory conditions. Blocking MMP15 activities suppresses collagenase, gelatinase, and stromelysin in corneal injury to preserve the integrity and architecture of the stroma as wound healing progresses.

Candidate: Neda Heydarian

Poster #: 41

IL-7 and IL-15 Support Long-Term Survival of Memory Th17 Cells by Suppressing Their Death in a Model of Chronic Dry Eye Disease

Neda Heydarian, Seokjoo Lee, Yeganeh Farsi, Rohan Bir Singh, Yihe Chen, Reza Dana

Purpose: To evaluate the anti-apoptotic effects of IL-7 and IL-15 on the survival of pathogenic memory Th17 (mTh17) cells in a murine model of chronic dry eye disease (DED).

Methods: Female IL-17A-GFP reporter mice, aged 6 to 8 weeks, were placed in controlled environmental chamber for 14 days and then moved to a normal vivarium for the following 14 days to induce chronic DED. CD4+ T cells were isolated from the draining lymph nodes of chronic DED mice using magnetic-activated cell sorting. Subsequently, mTh17 cells were isolated by fluorescence-activated cell sorting. These cells were then stimulated with IL-2, anti-CD3, and anti-CD28 antibodies to induce activation-induced cell death (AICD). Either IL-7 or IL-15 (20 ng/mL) was added to assess their effect on limiting AICD in mTh17 cells. After 48 hours, cells were stained with Annexin V and 7-AAD to assess early (Annexin V+ 7-AAD-) and late (Annexin V+ 7-AAD+) apoptosis. Additionally, the expression levels of anti-apoptotic markers Bcl-2 (encoded by BCL2 gene) and BclxL (encoded by BCL2L1 gene), were assessed by RT-PCR, in IL-7 and IL-15-treated mTh17 cells post-AICD induction.

Results: Administration of IL-7 (p=0.0497) or IL-15 (p=0.0139) significantly increased the percentage of viable mTh17 cells compared to AICD-only conditions, with IL-7 restoring viable mTh17 cell counts to levels comparable to those in non-AICD controls. Notably, IL-7 significantly reduced early apoptosis in mTh17 cells (p=0.0475), while IL-15 had a lesser effect (p=0.018). However, IL-7 had only a moderate effect (p=0.378) on reducing late apoptosis in mTh17 cells post-AICD induction. Furthermore, IL-7 and IL-15-treated mTh17 cells showed significantly higher expression of anti-apoptotic genes Bcl2 (p=0.042) and Bcl2l1 (p=0.023).

Conclusions: These findings highlight the critical function of IL-7 and IL-15 in enhancing the long-term survival of mTh17 cells by increasing their resistance to apoptosis.

Candidate: Mark Krauthammer

Poster #: 42

Clinical Imaging and Immune Cell Population Differences in Corneal Graft Rejection Between Young and Adult Mice

Mark Krauthammer, Antonio Esquivel Herrera, Michael Bednar, Sunil Chauhan, Reza Dana, Thomas H. Dohlman

Purpose: Clinical and experimental evidence suggests reduced corneal graft survival following penetrating keratoplasty (PK) in pediatric patients. Graft rejection in this context may primarily involve any corneal layer, including the endothelium, stroma, and epithelium. However, the immunologic mechanisms underlying these age-related differences in rejection are not fully understood. This study aimed to compare clinical imaging features and quantify graft-infiltrating immune cell populations in a murine model of pediatric versus adult PK.

Methods: Corneal buttons from 10-week-old C57BL/6 mice were orthotopically transplanted into the right eyes of either 3.5-week-old (young) or 10-week-old (adult) male BALB/c recipient mice. Graft survival and rejection were monitored over eight weeks using slit-lamp biomicroscopy, optical coherence tomography (OCT), and in vivo confocal microscopy. These modalities were used to assess corneal opacity, thickness, reflectivity, and endothelial morphology. At days 7, 14, and 28 post-transplantation, corneal tissues were harvested for flow cytometric analysis. Leukocyte infiltration was quantified by measuring CD45⁺ cell frequencies, with further characterization of Th1, Th2, Th17 and Natural Killer (NK) cell subsets.

Results: There was a trend toward lower eight-week graft survival in young compared to adult recipients (20% vs. 40%, p = 0.19). Differences in rejection patterns were also observed between the age groups. Young mice predominantly exhibited a stromal fibrosis-like rejection phenotype, characterized by increased graft hyperreflectivity on OCT imaging. In contrast, adult mice demonstrated an edema-like rejection pattern, with hyporeflective graft appearances and greater graft thickness (adult mice: 266.7 ± 110.3 µm vs young mice: 218.5 ± 56.2 µm), although this difference was not statistically significant (p = 0.18). Confocal microscopy revealed that endothelial cell morphology was better preserved in young recipients. Flow cytometric analysis revealed marked leukocyte infiltration in both young and adult graft recipients at all time points, compared to naïve, age-matched controls. NK cell infiltration in the grafts showed a trend toward higher frequencies in young mice at days 7 and 14, though significance was only observed at day 7. Notably, NK cell activity assessed by the frequency of IFN-γ⁺ cells among NK populations (CD49b⁺IFN-γ⁺) was significantly elevated in young mice at day 28 (78.64 ± 6.8% vs. 46.17 ± 17.7%, p < 0.001). Th2 cell frequencies were significantly higher in young recipients at day 28 (6.06 ± 3.9% vs. 2.91 ± 1.4%, p = 0.03). Th1 cell frequencies were also significantly elevated in young mice at the same time point (14.89 ± 2.53% vs. 4.76 ± 2.32%, p < 0.001). No significant differences were observed in Th17 cell frequencies between the groups at any time point.

Conclusions: In a murine model of PK, we observed age-related trends in both clinical presentation and immune cell infiltration during corneal allograft rejection. Pediatric (young) mice exhibited a trend toward lower graft survival and demonstrated a fibrosis-like rejection phenotype, whereas adult mice more frequently showed an edema-like pattern suggestive of endothelial rejection. Immune cell profiling revealed increased NK cell activity and significantly elevated Th1 and Th2 responses in young recipients, suggesting a more robust alloimmune response. These findings underscore age as a potentially important factor influencing corneal graft rejection and may inform future efforts toward age-specific strategies to improve transplant outcomes, particularly in pediatric populations.

Candidate: Liangju Kuang

Poster #: 43

Mucoadhesive Micellar Eyedrops for the Treatment of Ocular Inflammation

Liangju Kuang, Yimin Gu, Yuting Zheng, Yavuz Oz, Ann Yung, Seokjoo Lee, Francesca Kahale, Reza Dana, Nasim Annabi

Purpose: Ocular inflammation is a leading cause of blindness worldwide. The current standard of care involves frequent administration of corticosteroid eyedrops over extended periods, resulting in poor patient compliance. This study aims to develop mucoadhesive eyedrops for sustained drug delivery, enhancing therapeutic efficacy and patient adherence.

Methods: Drug-loaded micelles (PBA-MC-LE) were synthesized by self-assembly of phenylboronic acid (PBA)-bearing block copolymers with loteprednol etabonate (LE, a corticosteroid) and incorporated into a hyaluronic acid-based matrix to form mucoadhesive eyedrops. LE concentrations were quantified using liquid chromatography. In vivo efficacy of PBA-MC-LE drops (1X/day) was compared to commercial LE drops (4X/day) and no treatment using a mouse model of electrocautery-induced inflammation. Corneal opacity was analyzed from slit lamp photos, central corneal thickness (CCT) measured via optical coherence tomography, corneal cytokine levels quantified by RT-PCR, and immune (CD45+) cell infiltration evaluated by immunohistochemistry. The biocompatibility was tested in a healthy mouse model.

Results: The in vitro drug release profile of PBA-MC-LE showed an initial burst release (51.9±3.9%) on day 1, followed by a sustained release for 12 days. In vivo, by day 7, commercial LE (83±11 µm, p=0.005) or PBAMC-LE (83±7 µm, p=0.002) drops treatment resulted in lower CCT compared to no treatment (125±73 µm). Additionally, LE (10.3±4.0%, p=0.002) or PBA-MC-LE drops (8.2±4.0%, p=0.0008) treatment reduced opacity areas compared to no treatment (45.4±34.4%). Interleukin (IL)-1β and IL-6 mRNA levels increased in untreated eyes compared to normal eyes (p<0.0001). However, treatment with LE (p=0.03, p=0.005) or PBAMC-LE (p=0.007, p=0.002) drops, significantly decreased IL-1β and IL-6 expression compared to no treatment. Furthermore, CD45+ cells in corneal epithelial and stromal layers were fewer with LE drops (p=0.003) or PBAMC-LE (p=0.005) treatment compared to untreated cornea. Micellar eyedrops were safe in mice.

Conclusions: The PBA-MC-LE (1X/day) drops demonstrated efficacy in treating ocular inflammation comparable to commercial LE drops (4X/day), presenting a promising platform for enhanced ocular drug delivery.

Candidate: Vinay Kumar Pulimamidi

Poster #: 44

Differentiation and Characterization of

Induced Pluripotent

Stem Cellderived Corneal Limbal Stem Cells

Vinay K. Pulimamidi, Olufemi S. Folorunso, Nima K. Dana, Jahnavi Bolleddula, Lei Xi, Nishant R. Sinha, Kanika Arora, Meghanashree M. Shreenivas, John M. Fallon, Sunil K. Chauhan

Purpose: Purpose:

Limbal stem cells (LSCs) play a crucial role in corneal tissue homeostasis. Autologous limbal grafts have been widely used to treat unilateral limbal stem cell deficiency (LSCD). However, for patients suffering from bilateral LSCD, induced pluripotent stem cells (iPSC)-derived LSCs could be a promising alternative to allogeneic limbal grafts. This study aimed to differentiate iPSCs into LSCs using defined culture conditions.

Methods: Human fibroblast-derived iPSC line was purchased from Wi Cell (Madison, WI). iPSCs were cultured on Matrigel-coated tissue culture dishes in mTeSR1 medium. The iPSC line was characterized by its expression of pluripotency markers (Oct4, Nanog, and SSEA4) and its capacity to form trilineage differentiation potential. Pluripotency was also evaluated by the teratoma formation assay in immunodeficient mice. The iPSCs were differentiated into LSCs using defined culture conditions. The iPSC-derived LSCs were characterized based on the expression of LSC-specific markers by qRT-PCR and immunofluorescence. Human leukocyte antigens (HLAs) expression was also evaluated by flow cytometry.

Results: The iPSC line expressed the pluripotency markers Oct4, Nanog, and SSEA4. The iPSCs were differentiated into the three germ layers and expressed ectoderm (OTX2), endoderm (SOX2), and mesoderm (Brachyury) specific markers. In the teratoma assay, we observed the presence of tissues from all three germ layers. At D9, pluripotency markers Oct4 (p=0.0002), and Nanog (p=0.0001) were significantly downregulated in iPSC-derived LSCs, while the eye-specific transcription factors PAX6 (p=0.0003) and p63α (p=0.0001) were significantly upregulated. On Day 24, the iPSC-derived LSCs showed increased expression of putative LSC markers PAX6 (p=0.0001), p63α (p=0.0001), ABCB5 (p=0.01), KRT 14 (p=0.0001), and KRT 15 (p=0.0001). Post-thaw iPSC-derived LSCs also retained the expression of LSC-specific markers. The expression of HLA molecules was maintained in iPSC-derived LSCs which was moderately lower compared to the corneolimbal epithelial cell line.

Conclusions:

Our data suggest that human iPSC-derived limbal stem cells could provide a promising alternative for the treatment of limbal stem cell deficiency. Further, preclinical proof-of-concept studies are warranted to explore their translational potential.

Candidate: Nikita Bagaev

Poster #: 45

Morphometric Analysis of Microglia in the Retina and Optic Nerve in Neurofibromatosis Type 1 Mice

Nikita Bagaev, Jonathan Soucy, Aubin Mutschler, Petr Baranov

Purpose: Microglia contribute to various neurological disorders, including the visual impairments observed in Neurofibromatosis Type 1 (NF1). In NF1, retinal ganglion cell (RGC) damage can be mediated by reactive microglia. Here, we performed a morphometric analysis of microglia in the retina and optic nerve of mice carrying an Nf1 (Neurofibromin 1) gene mutation to characterize reactive microglial states in NF1 by reconstructing their cellular morphology and tissue's 3D architecture.

Methods: We collected retinal whole-mounts and optic nerves from Nf1 flox/flox and Nf1 flox/- mice, staining them with the pan-myeloid marker Iba1 and nuclear stain DAPI. To enhance antibody penetration in dense optic nerve tissue, we used Cy3-conjugated VHH (heavy-chain–only fragments) secondary antibodies, and pre-treated samples with a permeabilization buffer. Samples were cleared using ScaleSQ and mounted with self-hardening ScaleSH media on concave microscope slides. Confocal 3D images were acquired with the Olympus Fluoview FV3000 (160 μm depth for retina, 200 μm for optic nerve, 2 μm steps, 20× objective). IMARIS (v10.1) was used for 3D surface reconstruction of microglia and process tracing via the Filament Tracer. Quantitative analysis and classification were performed using the Vantage module, based on metrics such as total cell area and process length.

Results: Our pipeline enabled precise characterization of microglial morphology and activation states (homeostatic, reactive, hyper-ramified, rod-shaped) in both retina and optic nerve. The integration of optimized tissue processing, high-resolution imaging, and AI-assisted analysis allowed accurate classification and quantification of dozens of morphometric features per cell (e.g., cell area, soma size, number and length of processes, Sholl analysis), all while preserving spatial integrity.

Conclusions: This scalable workflow provides a powerful framework for high-throughput morphometric analysis of microglia in retinal and optic nerve tissue. It is especially relevant for NF1 studies, where inflammation and immune response in the optic nerve contribute to disease progression. Furthermore, the protocol is adaptable to other image analysis applications in neuroscience and immunology.

Candidate: Hui-Chen Cheng

Poster #: 46

Gene Therapy Rescues Visual Function in Familial Dysautonomia

Associated Optic Neuropathy

Hui-Chen Cheng, Reynette Estelien, Caitlin Elizabeth Keiper, Yasaman Anvarinia, Matthew Chagnon, Luk H. Vandenberghe, Susan A. Slaugenhaupt, Anil Chekuri

Purpose: Familial dysautonomia (FD) is an autosomal recessive sensory and autonomic neurodegenerative disorder leading to complex neurological phenotypes. Optic neuropathy with progressive blindness is one of the most debilitating symptoms in these patients, which leads to severe visual impairment in the third decade of life and impacts patients’ quality of life. Currently, there is no treatment for vision loss in FD. Several therapeutic approaches have been previously shown to increase ELP1 in the retina and rescue RGC loss with varying success in preclinical studies. Here, we outline a gene supplementation strategy to elevate the human ELP1 protein in the retina as a potential therapy for optic neuropathy in FD. We developed a novel AAV vector (AAV2.U1a.hELP1) that can be specifically targeted to RGCs to restore normal ELP1 protein levels in the FD retina and significantly rescue optic neuropathy in a retina-specific knock-out mouse model of FD (Pax6Cre+;Elp1loxp/loxp).

Methods: To assess the therapeutic efficacy of AAV2.U1a.hELP1 treatment, we performed intravitreal injections of different doses of AAV2.U1a.hELP1 vector with either 5.4x108 viral genome (vg) or 2.7x109 vg in the Pax6-Cre+;Elp1loxp/loxp) mouse model (FD mice) and control littermates. Intravitreal injection of AAV2.eGFP with 4.75x108 vg was performed in control-treated eyes. Comprehensive structural and functional evaluation was performed using high-definition spectral-domain optical coherence tomography (SD-OCT), fullfield electroretinography (ffERG), flash visual evoked potential (VEP), pattern ERG (pERG), photopic negative response (PhNR), and optomotor response assay (OMR) . Continuous variables with normal distribution were analyzed by t-test or one-way ANOVA for group analysis. Kolmogorov-Smirnov test or Kruskal-Wallis test was used for continuous variables without normal distribution. A p<0.05 (two-sided) was defined as statistically significant.

Results: Our results indicated that FD mice had significant lower peripapillary RNFL thickness, lower amplitude of flash VEP, pERG, phNR and worse visual acuity and contrast sensitivity compared to control group. . In FD mice, the amplitude of flash VEP, pERG, phNR, dark-adapted a wave and light-adapted b wave on ffERG improved after intravitreal injection of AAV2.U1a.hELP1 with 5.4x108 vg, compared to non-injected ones. The visual acuity and contrast sensitivity also improved in FD mice after intravitreal injection of AAV2.U1a.hELP1. Intravitreal injection of AAV2.U1a.hELP1 did not result in significant changes in our control mice.

Conclusions: This is the first study to evaluate the rescue of RGC function in the treatment of progressive optic neuropathy in FD. Significantly structural and functional defects in the retina-specific FD mouse model was observed. In addition, intravitreal injection of AAV2.U1a.hELP1 improved the visual function outcome in the FD mice compared to the control mice.

Candidate: Seonggyu Choe

Poster #: 47

Pilot Study of Head Scanning Behaviors of Drivers With Homonymous Visual Field Loss in Naturalistic Driving

Seonggyu Choe, Patrick Baker, Naser Al-Madi, Jina Yi, Chen-Yuan Lee, Shrinivas Pundlik, Gang Luo, Alex R. Bowers

Purpose: Prior studies evaluated head scanning behaviors of drivers with homonymous visual field loss (HVFL) either in driving simulators or in on-road driving tests. We have developed methods to study head scanning behaviors in naturalistic driving. We report early findings for two drivers with left HVFL focusing on frequently visited intersections in their daily routes.

Methods: Dashcams installed in personal vehicles recorded the driver and road ahead for S1 with left quadranopia (166 hours, 3,603 miles, 1964 intersections) and S2 with left homonymous scotoma (74 hours, 2,525 miles, 2,120 intersections). Using a custom tool, intersections were identified based on GPS location, and frequently visited intersections were identified for analysis. Video clips corresponding to those intersections were annotated for intersection characteristics, including geometry, traffic signs, environmental conditions, and traffic volume. Head scans, defined as yaw rotations exceeding 20° for at least 167 ms to the left or right of straight ahead, were analyzed before and within intersections.

Results: Both drivers showed consistent scanning strategies: a left-right-left pattern when driving straight through an intersection and a right-left-right pattern when turning right. Across all intersections, leftward scans were more frequent than rightward scans (S1: 3.0 vs 1.5 per intersection, p < .001, S2: 1.6 vs 1.3, p = .019) and had larger magnitudes (S1: 48.5° vs. 38.9° p < .001, S2: 50.3° vs 33.6°, p < .001). For S2, the presence of cross traffic on the non-seeing (left) side modified scanning behaviors on right turns with a stop sign: leftward scan rates (the proportion of right turn maneuvers with leftward head scans) were higher when left-side cross traffic was present (90% vs. 34%) and leftward scan magnitudes were larger (69.1° vs. 61.8°). Surprisingly, adverse environmental conditions (e.g., rain) reduced overall scan rates (S1: 23% to 11%, S2: 26% to 17%).

Conclusions: Preliminary results suggest that HVFL drivers exhibit compensatory head-scanning behaviors in their daily driving, making more scans with greater magnitudes toward their non-seeing side, as reported in some on-road and driving simulator studies. Naturalistic driving recordings provide additional insights into how environmental conditions and traffic volume may modify these scanning behaviors. Data collection is ongoing.

Candidate: Maria Emfietzoglou

Poster #: 48

Inner Plexiform Layer Substrata are Discernible With Commercial OCT and Affected by Aging

Maria Emfietzoglou, Victor S.M.C. Correa, Gustavo Sakuno, Rosanne Naafs, Joan W. Miller, Alexander Charonis, Demetrios G. Vavvas

Purpose: This study aims to evaluate the inner plexiform layer (IPL) microstructure and its changes with aging using commercial spectral-domain optical coherence tomography (SD-OCT) macular scans of healthy individuals with a semi-automated segmentation program.

Methods: Cross-sectional study conducted at the Athens Vision Eye Institute from January to July 2024. The study included 92 healthy participants. OCT images were captured with the Optovue Avanti SD-OCT and processed using ImageJ/FIJI to measure thickness and analyze the hyperreflective and hyporeflective bands within the IPL. Information about signal intensity, microstructure, and contrast between these sublayers was obtained. Statistical analyses, including Spearman’s correlation and linear regression, assessed relationships between age and IPL extracted features. Intra- and inter-eye repeatability were evaluated using paired samples t-tests combined with bootstrap analyses. The primary outcomes measured were signal intensity of the IPL, contrast between its hyperreflective and hyporeflective bands, and the percentage of IPL with identifiable sublayers. Secondary outcomes included inner retinal thickness measurements, including the IPL, nerve fiber layer (NFL), and ganglion cell complex (GCC).

Results: The IPL exhibited a multi-layered structure with five sublayers, three hyperreflective and two hyporeflective, arranged in an alternating pattern. Aging was associated with higher signal intensity from hyporeflective bands and minimal changes in hyperreflective bands, resulting in an overall reduced contrast between the five sublayers. Older participants showed a lower percentage of IPL with identifiable sublayers, along with a lower contrast variance within the IPL. Aging also correlated with reduced inner retinal thickness, including the IPL, NFL, and GCC, with a stronger association for the IPL. IPL analysis exhibited high intra- and inter-eye repeatability, with significant correlations and non-significant mean differences observed in most key parameters.

Conclusions: Analysis of the IPL and its sublayers is both feasible and reproducible using commercially available OCT along with a semi-automated segmentation program. Our findings indicate that the IPL microstructure changes with aging. A comprehensive evaluation of the IPL could serve as a valuable biomarker for early diagnosis and monitoring of diseases affecting synaptic health in this layer.

Candidate: Sierra Ha

Poster #: 49

Deep-Learning Based Orbital Volumetric Analysis of Teprotumumab Response in Thyroid Eye Disease

Sierra K. Ha, Lisa Y. Lin, Adham M. Alkhadrawi, Synho Do, Nahyoung Grace Lee

Purpose: Thyroid eye disease (TED) involves complex interactions between autoantibodies, the insulin-like growth factor-1 receptor (IGF-1R), and downstream inflammatory mediators, resulting in proliferation of orbital fibroblasts. Teprotumumab, an IGF-1R inhibitor, has emerged as a promising treatment by modulating these pathways. While quantification of orbital structures has historically relied on labor-intensive manual measurements with inherent variability (1-3), we previously published a deep learning-based automated segmentation model to accurately assess orbital muscle and fat volumes on orbital computed tomography (CT) in TED patients (4). Here, we apply this model to evaluate volumetric changes in orbital muscle and fat following teprotumumab and assess correlations with clinical outcomes.

Methods: This retrospective study included TED patients treated with teprotumumab who had pre- and posttreatment CT imaging of the orbits over a five-year period at a single institution. Orbital muscle and fat volumes were measured using a deep learning automated segmentation model based on a 2D U-Net architecture previously developed by our group. Primary outcomes included changes in orbital muscle and fat volumes, assessed using paired t-tests. Mixed-effects multivariable regression, adjusting for age, sex, and smoking status, was performed to investigate the relationship between volumetric changes and clinical parameters, including proptosis, diplopia, and clinical activity score (CAS). In regression models, a random effect for each participant was included to account for the potential correlation between two orbits from the same individual.

Results: Forty-four orbits from 22 patients (13 female, 9 male) were included. Volume of each of the four rectus muscles and total orbital muscle volume significantly decreased after treatment (all p < 0.01), whereas fat volume showed no significant change. Decreased muscle volume was observed in 39 orbits (88.6%) from 20 patients following treatment with teprotumumab, with an average reduction of 23.0% ± 15.6% (p < 0.01). Conversely, 5 orbits from 3 patients demonstrated increased muscle volume post-treatment. Decreased muscle volume was significantly associated with reductions in proptosis (p < 0.01). Changes in both muscle and fat volumes were not significantly associated with diplopia or CAS.

Conclusions: This deep learning-based study found significant extraocular muscle volume reduction after teprotumumab, consistent with studies using manual segmentation methods but with enhanced efficiency (~30 seconds per scan) (2). Notably, there were no significant changes in fat volume. This analysis revealed a correlation between muscle volume reduction and proptosis improvement, a relationship not observed in an earlier study using manual cross-sectional area measurement (1). This study underscores the potential utility of automated volumetric analysis in offering rapid, objective assessment of treatment response and optimizing clinical management of TED. These findings reinforce the value of incorporating advanced imaging and computational techniques in personalized treatment strategies for TED.

Candidate: Yi Ni Toh

Poster #: 50

Novel Head Scanning Training Improves Blind-Side Scanning and Hazard Detection in Drivers With Hemianopia

Yi Ni Toh, Patrick Baker, Jing Xu, Alex R. Bowers

Purpose: Driving is crucial for independence, yet hemianopia (loss of half the visual field) significantly compromises drivers' ability to detect hazards on their blind side. This deficit is particularly dangerous at intersections, where large scans with head movements are necessary to view the entire cross-street. While compensatory scanning is possible, many individuals with hemianopia fail to scan adequately, increasing collision risk. We conducted an open-label evaluation of a novel intervention which uses auditory reminder cues to train proactive blind-side scanning on approach to intersections.

Methods: Fifteen participants with hemianopia completed three training sessions in a high-fidelity driving simulator equipped with eye and head tracking. They drove along city routes with many intersections and other traffic. If a participant failed to make a large blind-side head scan when approaching an intersection, the system emitted a directional beep from a loudspeaker on that side, serving as a reminder to scan. The primary goal was for participants to scan early and thereby avoid these reminder beeps. To quantify training effects, scanning behaviors and hazard detection performance were compared using data from driving simulator evaluations before training, 1-week, and 1-month after training. These evaluation drives included hazardous motorcycle events at 38% of the intersections, requiring a horn press upon detection, and importantly, did not provide any reminder beeps. Detection tasks were not included in the training.

Results: Training significantly improved blind-side scanning and hazard detection, with improvements maintained after a month. Pre-training, participants made an early large blind-side eye-with-head scan at only 25% of intersections, increasing significantly to 90% at 1-week post-training and 80% 1-month later (ps < .001). Blind-side motorcycle hazard detection improved from 56% pre-training to 85% (1-week) and 83% (1month post-training) (ps < .001). This was driven by more effective blind-side scanning, with scanning-related misses (failing to scan and not scanning far enough) decreasing significantly from 39% to 13% (1-week) and 15% (1-month). Seeing-side detection remained high (>92%).

Conclusions: Reminder-cue training successfully fostered a proactive scanning style (more numerous, larger, earlier blind-side head scans before intersections), leading to significantly improved blind-side hazard detection that persisted for at least 1 month. This open-label clinical trial provides initial evidence that auditory remindercue training is a promising approach to rehabilitate driving-related hazard detection deficits in individuals with hemianopia.

Candidate: Ayush Parikh

Poster #: 51

Periorbital and Facial Necrotizing Fasciitis in an Immunocompetent

Adult

Ayush Parikh, Sri Meghana Konda, Lisa Y. Lin, Nahyoung Grace Lee

Purpose: To highlight a rare case of periorbital necrotizing soft tissue infection (NSTI) in a healthy immunocompetent male.

Methods: A 37-year-old healthy male presented with a one-day history of rapidly progressing left periorbital and facial cellulitis. He reported active tobacco use but had no history of diabetes mellitus, immunodeficiency, or preceding trauma. Examination revealed edema and skin whitening of the left upper eyelid, and facial swelling extending to the ear and mandibular border. The patient was tachycardic and febrile, and labs showed a leukocytosis (25.1), elevated lactate (7.5), a negative HIV screen, and normal HbA1c. Computed tomography (CT) of the face revealed extensive scalp, periorbital, and facial cellulitis extending into the neck without a drainable abscess. The Laboratory Risk Indicator for Necrotizing Fasciitis (LRINEC) score was 6, supporting clinical suspicion for NSTI. The patient underwent emergency debridement and irrigation of the left upper eyelid, cheek and neck using saline, hypochlorous acid and antibiotics, performed by oculoplastic and plastic surgery teams. Intraoperative cultures were obtained and tissue samples were sent for histopathologic evaluation. He was admitted to the plastic surgery service and started on broad-spectrum intravenous antibiotics (vancomycin, cefepime, clindamycin, and metronidazole). On post-debridement day one, examination showed continued necrosis in the left upper eyelid but with early signs of tissue viability in the lower eyelid and improved facial swelling.

Results: Pathology demonstrated bacterial cocci within necrotic tissue, confirming the diagnosis of NSTI. Cultures grew Streptococcus pyogenes (Group A Streptococcus). Several days later, the patient reported new left brow pain and was subsequently reimaged, which revealed focal worsening in the temporal region requiring a 3rd debridement. The remainder of his clinical course was uncomplicated. By hospital day 19, he was transitioned to oral amoxicillin and discharged. Follow-up assessments indicated ongoing healing and preserved orbicularis function. Approximately 1.5 months after discharge, the patient underwent elective left upper eyelid reconstruction with a full-thickness skin grafting.

Conclusions: This case describes the importance of early recognition of necrotizing fasciitis even in the absence of traditional risk factors, as expeditious intervention and antimicrobial therapy are crucial. Streptococcus pyogenes was identified as the sole pathogen; monomicrobial NF is less common and typically affects healthy individuals. The pathophysiology of periorbital NF in healthy hosts remains unclear, but factors such as unnoticed trauma or occult infections may contribute.

Candidate: David Zhao

Poster #: 52

Active Smokers Undergoing Teprotumumab Treatment for Thyroid Eye Disease

May Experience Greater Improvement in Proptosis and Lid Position Than Non-Smokers

David

X. Zhao, Carolina A. Chiou, Tatiana R. Rosenblatt, Amee D. Azad, Darren A. Chen, Sierra Ha, Jia Jia Zhang, Lisa Y. Lin, Natalie

Wolkow, Nahyoung G. Lee, Michael K. Yoon, Suzanne K. Freitag,

Purpose: Smoking is associated with the development and progression of thyroid eye disease (TED) as well as diminished responses to immunosuppressive therapies. However, the impact of smoking on outcomes following teprotumumab treatment for TED remains poorly understood. This study aims to evaluate differences in treatment outcomes between active smokers and non-smokers who underwent teprotumumab therapy for TED.

Methods: This is a single-center retrospective cohort study of TED patients who completed an 8-infusion course of teprotumumab from January 1, 2022 to April 1, 2025. Outcomes of clinical activity score (CAS), diplopia (Gorman Score), proptosis of each eye (Hertel exophthalmometry), and margin reflex distance-1 (MRD1) of each eye were measured at the clinic visit prior to initiation of teprotumumab, first visit postteprotumumab, and most recent follow-up. Fisher’s exact test was used to compare distributions of CAS and Gorman Score between groups. Multivariable mixed effects models were used to compare exam outcomes in active smokers and non-smokers in the above three time points with random effects for patient and eye, and covariates for age, sex, race, prior orbital decompression, teprotumumab retreatment, and pre-teprotumumab exam. Inverse probability weighting using propensity scores from patient baseline characteristics was applied to mixed effects models to distribute the effects of confounders equally between groups.

Results: One-hundred sixty patients completed teprotumumab treatment, of which 12 (7.5%) were active smokers. Active smokers were more likely to have undergone prior orbital decompression surgery than nonsmokers (25% vs. 6.8%, p=0.026). All other demographic characteristics, pre-teprotumumab outcome measurements, and time between the three visits were not significantly different between groups. The median time between the last teprotumumab infusion and the most recent visit for all patients was 15 months (IQR 827). There was no significant difference in the distribution of CAS or Gorman Scores between active smokers and non-smokers at any timepoint. However, in multivariable ordinal logistic regression of pre-teprotumumab CAS, active smokers had a 2.67 times higher odds (95% CI 1.17-11.48) of having a higher CAS than nonsmokers. In multivariable mixed effects linear models, active smokers had a 1.6 mm greater reduction (95% CI –2.6, -0.6, p=0.002) in proptosis than non-smokers from the pre-teprotumumab visit to the most recent visit. There was significantly more reduction in MRD1 in active smokers compared to non-smokers from the preteprotumumab to the post-teprotumumab visit (-1.2 mm; 95% CI -1.9, -0.5, p=0.002) and to the most recent visit (-1.0 mm; 95% CI –1.8, -0.2, p=0.027).

Conclusions: TED patients that actively smoked throughout teprotumumab treatment and follow-up experienced greater long-term improvement in proptosis and sustained larger improvements in MRD1. However, active smokers had higher odds of presenting with more severe baseline inflammation than nonsmokers, suggesting that they may derive greater relative benefit from teprotumumab treatment. More longterm data is needed to understand TED re-flare after teprotumumab treatment in this subset of patients.

Candidate: James Harris

Poster #: 53

The Immune System in Health and Glaucoma

James Harris, Zhuoran Yin, Dario Tommasini, Paul Cullen, George Baldwin, Anthony Mukwaya, Nasir Uddin, Ryan Xue, Karthik Shekhar, Milicia Margeta

Purpose: Neuroinflammation is thought to play a role in the pathogenesis of many neurodegenerative conditions affecting the retina and brain, including glaucoma, a disease of progressive retinal ganglion cell (RGC) degeneration. To better understand the role of the immune system in glaucoma, we created a singlecell transcriptomic atlas of resident and invading immune cells in the optic nerve and retina from animals with glaucomatous degeneration from increased intraocular pressure and sham controls. Doing so allowed us to better understand the native immune state of these tissues and how it changes during RGC degeneration.

Methods: We used a murine microbead-injection glaucoma model. Wild type mice received either microbeads injection into the anterior chamber to inhibit aqueous outflow, increasing intraocular pressure (IOP) or sham saline injection. Increased IOP resulted in reproducible loss of retinal ganglion cells. Animals were sacrificed 30 days after treatment and retinas and optic nerves were harvested separately and pooled, creating 3-4 independent biological replicates per sample. These tissues were disassociated, immunostained and FASC sorted to isolate CD45+ leukocytes. scRNA sequencing libraries were prepared from sorted cells (10x genomics). Libraries were sequenced, demultiplexed, and aligned to create gene count matrices (CellRanger). Count matrices were analyzed in R with Seurat. Quality control was performed to exclude poor quality cells. Count matrices were normalized, scaled, principal component analysis was performed, cells were clustered and visualized with UMAP projections. Relative proportions of cell types were calculated for each condition and tissue and significant differences were tested with ANOVA. Differentially expressed genes were calculated using a pseudobulk approach with the DeSeq2 package. RNA sequencing findings were validated with immunohistochemistry performed on independent cohorts of control and microbead-injected mice as well as human post-mortem optic nerves using standard histological techniques and visualized with fluorescence microscopy.

Results: ~28,000 immune cells were sequenced from retinas and optic nerves in sham and microbead injected animals. Control animals had more activated microglia in the optic nerve than the retina, particularly at the nerve head. Immunostaining revealed a unique population of microglia at the nerve head tissue boundary in mice and humans. Elevated IOP significantly increased the abundance of “disease-associated” microglia, peripheral macrophages, and T-cells in the retina, but not the optic nerve. Many of the immunologic changes we observe in our glaucoma model are also found in models of outer retinal neurodegeneration, suggesting common mechanisms of retinal neurodegeneration.

Conclusions: Here we provide a comprehensive overview of the immunologic environment of the retina and optic nerve in healthy animals and how it changes in a model of glaucomatous neurodegeneration. We identify a novel microglial population at the optic nerve head in healthy animals. We also show significant expansion of microglia, peripheral macrophages and T-cells in the retina that accompany RGC degeneration. We also identified candidate immune molecules that may mediate degeneration and are potential future therapeutic targets. Many of these immunologic changes are also observed in other neurodegenerative disease models of the retina and brain suggesting potential common immune-mediated mechanisms.

Candidate: Asahi Fujita

Poster #: 54

Comparison

of Incidence and Risk Factors for

Glaucoma Surgeries

Among Pseudoexfoliation and Primary Open-angle Glaucoma: Analysis of IRIS® Registry

Asahi Fujita, David S. Friedman, Kanza Aziz, Alice Lorch, Joan Miller, Yan Zhao, Tobias Elze, Nazlee Zebardast, IRIS® Registry

Analytic Center Consortium

Purpose: To compare the incidence and predictors for glaucoma surgeries between pseudoexfoliation glaucoma (PXG) and primary open-angle glaucoma (POAG).

Methods: We identified patients with PXG and those with POAG by diagnosis codes recorded in 2015 using the IRIS® Registry (Intelligent Research in Sight). We balanced age, sex, race/ethnicity, baseline intraocular pressure (IOP), visual acuity (VA), cup to disc ratio (CDR), and lens status using nearest neighbor matching method. Survival analysis was used to compare the cumulative incidence of glaucoma surgery. Cox regression models were used to determine predictors of undergoing glaucoma surgery in each group including the following variables: age, sex, race/ethnicity, baseline IOP, baseline VA, baseline CDR, and lens status.

Results: 11,613 patients with PXG and 352,701 patients with POAG were identified, among whom 11,613 matched eyes from each diagnosis group were included in the survival analysis. The mean age was 77.4 (standard deviation: 9.3) years and 66% were females. The cumulative incidence rates of surgery were 15.7% [95% confidence interval (CI):15.0–16.4%] and 8.5% [8.0–9.1%] at 4 years in the matched PXG group and POAG group, respectively. Filtering surgery (trabeculectomy and tube shunt surgery) accounted for 51.3% and 39.7% of the surgeries performed for PXG and POAG, whereas minimally invasive glaucoma surgery (MIGS) accounted for 32.8% and 45.4%, respectively. Higher IOP, higher CDR, and male sex were associated with a higher likelihood of undergoing filtering surgery in both groups. In the POAG group, Black individuals (vs. White, hazard ratio:1.16 [95%CI: 1.10–1.22]) were more likely to undergo filtering surgery, while in the PXG group, White individuals were (vs. Black, 2.12 [1.38–3.25]). Factors associated with a higher likelihood of undergoing MIGS in both groups included age 60–79 years, higher IOP, and phakia. In the POAG group, being White (vs. Black, 1.18 [1.12–1.25]) was also associated.

Conclusions: Patients with PXG were approximately twice as likely to undergo surgery as those with POAG, particularly filtering surgery. The likelihood of undergoing surgery varied by race depending on the diagnosis.

Candidate: Yan Luo

Poster #: 55

Improving Instructional Compliance for AI-Generated Fundus Photos With Demographics-Aware Diffusion Model

Yan Luo, Minghan Li, Mohammad Eslami, Saber Kazeminasab, Tobias Elze, Lucy Q. Shen, Louis R. Pasquale, Nazlee Zebardast, Michael V. Boland, David S. Friedman, Mengyu Wang

Purpose: This study investigated how to improve instructional compliance in text-to-image generation for fundus images by demographics-aware modeling.

Methods: We conducted a retrospective analysis of 10,000 Scanning Laser Ophthalmoscopy fundus images obtained from 10,000 distinct patients who received care at Massachusetts Eye and Ear between 2015 and 2022. The sample population (mean age: 61.6 ± 15.6 years) comprised three racial categories: Asian (8.2%), Black (14.9%), and White (76.9%). Gender distribution included Female (58.2%) participants, while ethnic composition consisted of Hispanic (3.8%) and non-Hispanic (96.2%) individuals. To ensure robust model evaluation, we implemented a stratified partitioning strategy: training (60%), validation (10%), and test (30%) sets. To address demographic biases in image generation, we developed FairDiffusion, a novel equity-aware diffusion model incorporating Fair Bayesian Perturbation. Conventional stable diffusion models were used as baselines, using prompts constructed by incorporating relevant demographic attributes and disease labels, e.g., a fundus image of a white female non-Hispanic glaucoma patient. Evaluation metrics included the Area Under the Curve (AUC), Group-wise AUC, and the Equity-Scaled AUC (ES-AUC). Higher ES-AUC values indicate more equitable performance across groups.

Results: We evaluated text-to-image generative models' instruction compliance by training a classifier on generated fundus images with their text labels (e.g., "glaucoma") and testing their performance on real data. Higher AUC indicates better alignment between generated images and instructions. Quantitative analysis showed that FairDiffusion achieved an overall AUC of 0.68 while the overall AUC of Stable Diffusion (FT) was 0.61, gaining a 0.07 improvement (p<0.001). Consistent improvements could be observed in ES-AUC: gender ES-AUC improves from 0.59 to 0.66 (+0.07), race ES-AUC rises from 0.57 to 0.63 (+0.06), and ethnicity ESAUC advances from 0.58 to 0.64 (+0.06). These improvements extended to underrepresented demographic subgroups as well, with improvements in AUC for females (from 0.59 to 0.67), Black (from 0.61 to 0.66), and Hispanics (from 0.56 to 0.63).

Conclusions: These findings demonstrate that FairDiffusion is an effective generative AI paradigm that enhances model performance and promotes demographic equity.

Candidate: Mousa Moradi

Poster #: 56

Glaucoma Subtype Detection From Unstructured Clinical Notes Using Semi-Supervised Generative Adversarial Networks Across Ethnic Groups

Mousa Moradi, Rishi Shah, Asahi Fujita, Daniel Liebman, Kanza Aziz, Saber Kazeminasab, Mohammad Eslami, Mengyu Wang, Tobias Elze, Nazlee Zebardast

Purpose: To enhance Glaucoma subtype classification for underrepresented populations by utilizing unstructured Electronic Health Record (EHR) data

Methods: The MEE dataset includes 117,695 unique patients and 6,248,595 total clinical notes across various glaucoma subtypes such as POAG, ACG, NTG, NVG, and PXG. For the initial stage, we utilized 838 labeled notes and a pool of 1.5 million unlabeled notes from 60,000 patients. Given the limited size of the labeled set, we fine-tuned Low-Rank Adaptation of Large Language Models (LoRA), which allows efficient training by reducing the number of trainable parameters to 0.8%. This approach is especially useful for models trained on small, labeled datasets. LoRA was applied to the ClinicalBERT model, a pre-trained transformer specifically designed for medical text. Data preprocessing involved de-identification, stop word removal, and augmentation techniques to optimize the input. The SGAN-LM consists of a generator and a discriminator. The generator introduces Gaussian noise to the ClinicalBERT-embedded notes, simulating "fake" notes. The discriminator learns to distinguish between these generated fake notes and real labeled notes, iteratively improving both components. To handle the memory challenges posed by the large number of unlabeled notes, we used a subset of 60,000 patients, balancing computational efficiency and dataset diversity.

Results: The presented results demonstrate the robust performance of our model across racial groups in predicting GL and its subtypes, achieving an overall accuracy of 98.31%, precision of 96.51%, recall of 96.51%, and F1-score of 96.50%. Our results depict the ROC curves for White, Black African American, and Asian populations, with AUC scores ranging from 82% to 99%. Our results show case confusion matrices for the same groups, highlighting accurate predictions across six classes: Non-GL, POAG, PACG, PDG, XFG, and Other. The AUCs for Class 2 (PACG) and Class 3 (PDG) consistently exceeded 95% and 90%, respectively, indicating the model’s ability to distinguish these classes from other GL subtypes.

Conclusions: Our proposed SGAN model could successfully distinguish five glaucomatous subtypes (POAG, PACG, PDG, XFG, and Other) in EHR notes, achieving high accuracy, precision, recall. These findings demonstrate that an SGAN model can leverage and relatively small, labeled dataset and large unlabeled data to improve diagnostic performance, enhance generalization, and mitigate the risk of overfitting and underfitting in GL classification.

Candidate: Prashit Parikh

Poster #: 57

Eye Care Utilization Among Glaucoma Suspects: An IRIS Ⓡ Registry

(Intelligent Research in Sight) Analysis

Prashit Parikh, Asahi Fujita, James D. Brandt, Elizabeth Ciociola Kelly, Alice Lorch, Joan Miller, Tobias Elze, Nazlee Zebardast

Purpose: To investigate eye care utilization patterns of glaucoma suspect patients in the U.S.

Methods: Using the American Academy of Ophthalmology IRIS® Registry (Intelligent Research in Sight), we identified patients with a diagnosis code for glaucoma suspect between 01/01/2015 and 02/28/2020. We excluded anyone with glaucoma diagnosis codes, history of glaucoma-related procedures, intraocular pressure (IOP) >=24 mmHg, cup-to-disc ratio (CDR) >0.6, and CDR asymmetry (difference >=0.2) during a one-year lookback period. Patients were stratified into three groups: 1) low-risk: IOP 5-23 mmHg, CDR <0.6, and no CDR asymmetry; 2) ocular hypertension (OHTN): IOP >=24 mmHg, CDR <0.6, and no CDR asymmetry; 3) high-risk: high CDR (either asymmetry or CDR 0.6-0.74) and any IOP. Outcome measures included the number of eye exams, visual field tests (VFs), and optical coherence tomography tests (OCTs) performed per patient per year, as well as glaucoma medication prescription rates. Demographic characteristics were collected for all patients and used to analyze care utilization across race and ethnicity groups. Among our cohort, 73.7% of patients were non-Hispanic White, 8.6% non-Hispanic Black, 6.7% Hispanic, 2.3% Asian or Native Hawaiian or Pacific Islander (A-NH-PI), and 0.3% American Indian or Alaska Native.

Results: A-NH-PI patients received the most eye exams in all groups and most OCTs in the OHTN group. Non-Hispanic Black patients underwent VFs most often in all groups, and the most OCTs in the low-risk group. Non-Hispanic White patients underwent the most OCTs in the high-risk group. 17.6% of all patients were prescribed medication during the analysis period with non-Hispanic Black (23.8%) and Hispanic (20.9%) patients having the highest prevalence. The OHTN group was prescribed medication most often (28.5%). Overall, patients prescribed medication (3.0 eye exams, 0.76 VFs, 1.23 OCTs) had higher utilization rates than those who were not (1.93 eye exams, 0.42 VFs, 0.82 OCTs).

Conclusions: Across 142,718 patients (mean age: 67.5 years), glaucoma suspects received on average 2.13 eye exams, 0.48 VFs, and 0.89 OCTs. The low-risk group had the lowest utilization rate for all outcome procedures. Eye exam (2.49) and OCT (1.05) utilization was highest in the OHTN group while VFs (0.56) were most often performed in the high-risk group. Overall, glaucoma suspect patients have high eye care utilization with OHTN and high-risk CDR features being associated with more frequent testing. Medication prescription was associated with increased eye care utilization in all groups.