NEI - National Eye Institute Grants

NEI - National Eye Institute

Project Summary/Abstract Myopia is the most common eye condition worldwide, where distant objects appear blurry. Typically, myopia develops during childhood/adolescence but increases the risk of developing vision-threatening conditions, including macular degeneration, retinal detachment, and glaucoma later in life. Furthermore, uncorrected distance refractive error has been estimated to result in a global loss of productivity of $202 billion annually. The rapid increase in myopia prevalence also known as the “Myopia Boom” is alarming, but existing treatments are only partially effective or induce significant adverse effects. We have discovered that indigo light can completely suppress a strong myopiagenic stimulus in a near-primate model. Based on extensive preclinical research, we have identified the most effective indigo wavelengths for the suppression of myopia. Importantly, our discovery is consistent with the environmental origin of the Myopia Boom, where sunlight, with its abundant indigo photons, is protective and time spent indoors, where indigo photons are mostly absent, is a risk factor. Based on this discovery, we propose a new modality for the treatment of myopia progression in children and adolescents: indigo light emitting eyewear. This is an exciting treatment strategy because it may enable a highly effective, safe, noninvasive, simple to use, and cost-effective treatment for childhood myopia. In this Phase I project, we propose (i) to advance technical merit by developing a new prototype for low volume production and formal safety evaluation; and (ii) to evaluate feasibility by performing a pilot clinical study in young adults assessing tolerability, usability and safety. If successful, we will have established technical merit and feasibility for a fully powered clinical trial to assess efficacy in myopic children.

NEI - National Eye Institute

PROJECT SUMMARY / ABSTRACT Ocular disease may arise anywhere in the human retina, which spans roughly 70% of the eye's internal surface. While optical coherence tomography (OCT) has become the premier technology for retinal disease diagnosis, current instruments are incapable of accessing the peripheral retina due to slow imaging speeds. This proposal seeks to enable next-generation OCT technology that will make diagnostic screening and visualization of both the central and peripheral retinal areas a reality; increasing imaging speed roughly 50-fold will allow wide-field retinal imaging without increasing procedure time. This advancement is empowered by a novel laser technology, stretched-pulse-mode-locking, recently developed for OCT by the founders of Bluebird Photonics. This Phase I STTR support will allow Bluebird to refine the SPML laser design and advance from a prototype to a turn-key, environmentally robust commercial laser. Specific aims include the implementation of a bias-free, Sagnac-loop amplitude modulator and the development of a closed-loop locking mechanism to ensure a stable synchronization between the modulator and the optical cavity resonance. The resulting laser will offer a significant advance over existing commercial OCT technology in speed (5 MHz vs ~100 kHz), cost, stability, reliability, and ease of integration. Commercial SPML lasers will serve as a catalyst for further development of ultrawide-field OCT instrumentation and support clinical evaluation across a range of diseases.

NEI - National Eye Institute

PROJECT SUMMARY Global Ophthalmology is an expanding field within ophthalmology that unifies the local and international community to advance research, clinical care, education, and policy to prevent blindness worldwide. The Global Ophthalmology Summit is the only United States (U.S.) based meeting that delivers in depth and focused content in research in global eye health and health equity. It brings together leaders to create community and foster collaboration amongst meeting participants which is an opportunity to captivate a young audience to lay the foundation for future collaborations, mentorship, and inspire a career in vision science. Key research topics addressed include global scale collaborative research consortia, disease-specific interventions, technology and innovation, sustainability, education, and advancing eye health systems in the U.S. and globally. The resources sought in this proposal aim to enhance the participation of young researchers within the field of global ophthalmology through the following aims. In aim 1 we want to provide an opportunity to strengthen the global eye research community by bringing local and international expertise together to focus on innovative solutions to prevent and treat blindness worldwide. In aim 2 our goal is to create collaboration through providing intentional meeting content amongst the diverse group of attendees. In aim 3 we want to support innovative vision research in global ophthalmology of trainees and early career investigators. The mission of the Global Ophthalmology Summit is in alignment with the National Eye Institute’s new strategic plan to drive innovative vision research, foster collaboration, build global research consortia, inspire and recruit a diverse workforce, and educate our community and policymakers on the pressing needs of vision research. The National Eye Institute’s cross cutting areas of research emphasis addressed in past and future Summits include genetic research, data science, individual quality of life, and public health and disparities research.

NEI - National Eye Institute

This proposal seeks support for students and junior investigators (travel awardees) to attend the 2025 International Society for Eye Research (ISER)/BrightFocus Foundation (BFF) glaucoma meeting titled "Concepts and Breakthroughs in Glaucoma" to be held October 8th-11th, 2025 at the Emory Conference Center and Hotel in in Atlanta, Georgia. As in past meetings, our goal is to bring together basic scientists, clinician-scientists, students and fellows for presentations and in-depth discussions on recent exciting research advances and developments in the molecular mechanisms underlying glaucomatous pathology, both in the conventional outflow tract and the optic nerve head. We have already recruited three thought leaders in glaucoma to deliver keynote lectures. As in past meetings, platform sessions will be selected exclusively from submitted abstracts, with one session reserved for travel awardees. We are also organizing again a one-day "crash course" in glaucoma for people newcomers to the field and students/junior investigators, consistent with our goal of increasing young scientist participation. The Specific Aims of the conference are to: 1. Enhance the emerging careers of at least 30 young investigators working in glaucoma research by providing travel awards. 2. Provide a forum for the dissemination of the most recent advances in glaucoma research. 3. Create an environment that facilitates the exchange of novel ideas among basic and clinician-scientists, fostering opportunities for collaboration among vision scientists with multiple scientific expertise. 4. Bring together scientists working in disparate areas of glaucoma research. We anticipate that this meeting will provide state-of-the-art information on recent advances in glaucoma and serve as an important resource for those involved in the translation of these findings into novel therapeutics. The conference will also provide new opportunities, avenues for new discovery, and a forum to develop potential collaborations among the attendees. The requested funds will support the travel, accommodation and registration of at least 30 trainees to attend this focused meeting.

NEI - National Eye Institute

Dry Eye Disease (DED) affects 20% of the global adult population, with higher rates in developed countries like the USA. It significantly impacts quality of life, productivity, increases healthcare costs, and can lead to vision loss if untreated. Current therapies are incapable of addressing chronic DED as they cannot be used long-term due to their toxicity. Our innovative First-In-Class mitochondrial technology targets the root cause of DED by addressing oxidative stress and mitochondrial dysfunction, which induces Meibomian Gland Dysfunction (MGD), a predominant cause of DED. Due to its unique mechanism of action, our drug candidate is safe for long-term use and offers a broader therapeutic window. This noninvasive treatment promises to revolutionize eye care and provide a sustainable solution to this widespread health challenge. Our proprietary small molecule lead candidate exhibits dose dependent free radical scavenging potential with calculated IC50 of 2.1µM and was found to protect various ocular cell lines against oxidative stress. It was found to be highly efficacious in reducing Benzalkonium chloride induced dry eye, tissue damage and vascularization in mice model of dry eye. An ophthalmic safety study suggests that an acute TID dose of our candidate as eye drops up to 30 µg/eye (total dose of 90 µg/eye/day) was well tolerated in New Zealand White rabbits. We have secured our invention by the filing of patent applications in the USA (US2021163159081), China (CN2280008594), India (IN202317001652), Canada (CA3205099A1), EU (EP22767950.3), Brazil (BR 1120230181097), Australia (AUS 2022234307) and Japan (JP2023-555226) for compounds, compositions, and their method of use. The novelty of our invention is confirmed by a written opinion from USPTO and an enhanced freedom to operate searches by qualified patent attorneys. DED is a multifactorial indication; therefore, in proposed SBIR phase I, our proposed studies entail In vivo characterization of lead candidates in desiccating stress/scopolamine (DSS) model of DED in mice working through varied etiologies. We also propose to evaluate the toxicity of our lead in repeated dose 7-day rabbit model of toxicity with PK parameters and local biodistribution in eye tissues of rabbits. Successful completion of proposed studies will be followed by submission of a phase-II SBIR to conduct a set of IND-enabling pre-clinical studies, manufacturing process scaleup and development, and analytical method validation, GLP toxicity studies with recovery group. Setup of in-house eCTD software interface and electronic submission gateway (ESG) with FDA and relevant certifications of personnel and workstation infrastructure will also be included in the SBIR phase-II application. We have assembled an experienced team consisting of chemists, drug discovery and bench to bedside drug product development experts, pharmacologists, ocular pharmacokinetics, toxicologists, ophthalmologists, regulatory experts, and industry partners. We are confident of successfully completing the proposed work.

NEI - National Eye Institute

Project Summary The 2026 Gordon Research Conference (GRC) on Photosensory Receptors and Signal Transduction (PRST) will be held in Ventura CA. The conference incorporates leading experts on diverse photosensory systems allowing for a comprehensive understanding of how nature employs cofactor chemistry to allow organisms to sense and adapt to their lighting environment. A comprehensive understanding of photoreception and signal transduction has a wide impact on human health and disease. For instance, for vertebrate vision subtle defects within the phototransduction pathway impacts retinal development and retinal degeneration, leading to conditions such as photophobia, visual acuity, night-blindness, loss of vision, as well as numerous other disorders of vision. Notably, defects in non-visual photo responsive systems within the eye have also been implicated in mood and sleep disorders, circadian dysfunction, and pupillary responses. To address these issues, it is essential to dissect and delineate how subtle alteration in cofactor identity, protein structure, and signaling pathways impacts defects in signaling and/or facilitates evolutionary adaptation to unique lighting environments. As a result, the format of the conference is unique in three essential elements: 1) Invited speakers will cover the entire range of physical, chemical, and biological factors gating light-responsive signal transduction in organisms. The comprehensive focus beginning from initial photon absorption, changes in cofactor chemistry, and structural transitions in photoreceptor proteins, to changes in organism physiology and ecological impacts affords unprecedented insight into how organisms harness photon absorption to mediate physiological responses. 2) Invited speakers cover the entire range of natural photoreceptors, providing deep understanding of how nature tunes small molecule cofactors, protein structure, and signaling pathways to allow for exquisite sensitivity to a wide range of wavelengths and light intensities. 3) These elements allow for an additional focus on applied practical applications to harness or manipulate photoreceptor function through optogenetic tools, or interventional treatments. The program structure and invited speakers for the GRC and associated Gordon Research Seminar (GRS) are designed to maximize interaction between established investigators, new investigators, and trainees from diverse disciplines to foster an exchange of ideas and scientific viewpoints to catalyze new interdisciplinary approaches to delineate photosensory responses and to engineer new technologies. The GRS provides an opportunity for junior scientists to meet and present their work in an environment of peers prior to the GRC. The GRS incorporates social, scientific, and mentoring to develop a sense of belonging, community, and inclusivity. Mentoring sessions have been chosen to encompass both academic and industrial career pathways to facilitate career growth and networking opportunities. The thoughtfully coordinated PRST GRC/GRS will position researchers to address the field’s most critical scientific challenges, while also motivating trainees to leverage their expertise in pursuit of emerging frontiers in photochemistry and photobiology.

NEI - National Eye Institute

Project Summary Cephalopod neuroscience offers a unique perspective in comparative brain research. Similar to vertebrates, cephalopods have evolved large, complex brains, enabling remarkable sensory, motor, and cognitive abilities. They exhibit sophisticated behaviors such as independent control of eight flexible arms, dynamic skin patterning for camouflage, and advanced learning and decision-making capabilities. Understanding the cephalopod nervous system has the potential to uncover fundamental principles of brain organization and function across species. Despite their fascinating neurobiology, the mechanistic workings of cephalopod brains remain largely unexplored. However, recent technological advances have catalyzed rapid progress and an influx of new researchers into the field, leading to the establishment of the first Cephalopod Neuroscience Gordon Research Conference. This meeting will bring together scientists from diverse areas of cephalopod research, including genomics, neural development, systems neuroscience, computation, and tool development. Our key objectives are to: (1) foster knowledge exchange and highlight recent discoveries, (2) cultivate an engaged and collaborative research community, and (3) facilitate resource and technique sharing. A strong emphasis will be placed on supporting trainees to ensure broad participation, and provide a strong foundation for this new Gordon Conference in the future. By combining cutting-edge science with community-building efforts, this conference aims to accelerate advances in cephalopod neuroscience and provide insight into broad principles of brain function.

NEI - National Eye Institute

Abstract Age-related macular degeneration (AMD) and related macular dystrophies (MDs) are leading causes of adult blindness with limited treatment options. AMD/MDs can present in two forms, geographic atrophy/GA (dry form) and choroidal neovascularization/CNV (wet form). There is strong evidence linking sterile inflammation to AMD/MD pathogenesis and two complement pathway inhibitors are already approved by FDA for treating GA in the dry form of AMD. However, due to limited therapeutic impact and adverse effects of complement inhibitors and other approved drugs for both dry AMD and wet AMD, there is a significant need for novel therapies for AMD/MDs. Our recently published data and preliminary studies identified secretory phospholipase A2-IIA (sPLA2-IIA), a pro-inflammatory enzyme, as a key molecular player in AMD/MD pathogenesis. AMD/MD primarily affect the retinal pigment epithelium (RPE) cells in the eye and patient-derived induced pluripotent stem cell- RPE (iRPE) from AMD and 2 distinct MDs showed elevated levels of sPLA2-IIA. Furthermore, AMD/MD iRPE cultures and AMD donor eyes showed elevated sPLA2-IIA levels in drusen, a pathological hallmark of early AMD/MD that is the key driver of later stage pathologies in AMD/MDs. Notably, pharmacological modulation of sPLA2-IIA activity in AMD and MD iRPE cultures led to reduced drusen. In addition, directly linking elevated sPLA2-IIA activity to AMD/MD pathology, sPLA2-IIA overexpression led to AMD-associated pathological alterations (drusen, Bruch’s membrane thickening, RPE thinning, CNV and visual deficits) in C57BL/6J mice. Altogether, these studies provide a strong rationale for targeting sPLA2-IIA activity in AMD/MDs. Toward this goal, we propose to develop proteolysis-targeting chimeras (PROTAC) compounds for specific inhibition of sPLA2-IIA in AMD/MDs. In initial experiments, we have synthesized a ‘lead‘ PROTAC (UR-00059) that can induce degradation of sPLA2-IIA in iRPE cells with half-maximal degradation concentration DC50 of 295.5 nM. The following milestone-driven aims will allow us to develop an effective PROTAC-based therapy targeting sPLA2-IIA for AMD/MDs. Aim 1: Optimize UR-00059 structure and activity and characterize the target engagement in vivo; Aim 2: Conduct in vivo efficacy studies and non-GLP absorption, distribution, metabolism, and toxicology of UR-00059; Aim 3: Perform IND enabling studies and obtain FDA approval for human testing. Ultimately, the proposed studies will develop a novel PROTAC-based therapy for targeting inflammation, drusen and consequently late stage pathologies of AMD and related MDs.

NEI - National Eye Institute

PROJECT SUMMARY/ABSTRACT This proposal seeks to fund and support trainees and junior faculty to attend the “Emerging Innovators in Ophthalmology Workshop” which will occur July 2026 at Stanford University with plans to make this a regular annual offering. Continued innovation in treatment, diagnosis and prevention of ophthalmic disease is critical for improving patient outcomes and reducing gaps in population health. Increasingly we recognize that innovation requires specific training and mentorship which is not broadly available. A specialized approach to evaluating clinical problems and creatively identify solutions has become codified and widely recognized around the world as the “Biodesign” approach upon which both the Stanford Byers Center for Biodesign Fellowship and, subsequently, the Stanford Byers Family Ophthalmic Innovation Fellowship Program are based. The “Emerging Innovators in Ophthalmology Workshop” will be an all-day course convened on Stanford campus to provide the framework of this approach and resources which attendees can use to further engage in the Biodesign process. Recognizing the importance of mentorship, senior faculty who both teach and engage in innovation will be present throughout the course and small group workshops to develop connections with the attendees. Additionally, each attendee will be connected with a formal innovation mentor based on their interest at the conclusion of the workshop with the expectation of regular meetings in the following year. Our objectives for the course are: (1) to teach the core principles of the Biodesign approach with a focus on Ophthalmic Innovation to ophthalmology residents, post-doctoral scholars, and junior faculty in hopes for encouraging innovation and (2) to provide and develop long term mentorship for clinical and scientific trainees as well as junior faculty in support of ongoing innovative work.

NEI - National Eye Institute

PROJECT SUMMARY The 2026 Gordon Research Conference (GRC) and Gordon Research Seminar (GRS) on Visual System Development are a paired set of biennial meetings that bring together investigators studying development, disease, and evolution of the visual system. Over the years, these meetings have provided an exciting and unique forum in which to explore the similarities and differences underlying visual system development and function across a broad range of species. The goal of these meetings is to foster an appreciation of common principles that mediate the construction and function of the visual system in diverse organisms, and to share the latest exciting new ideas and findings on this topic. By including sessions that highlight emerging topics with translational impact, such as “Retinal Stem Cells, Repair, and Regeneration”, and “Developmental Disorders, Diseases, and Aging of the Visual System”, the meeting is also expanding its scope and stimulating crosstalk between developmental biologists and investigators focused on translational aspects of vision science. The GRS provides a unique platform for students and postdoctoral research fellows in the visual research field to share current, unpublished research amongst their peers and receive career mentorship in the vision science field. The format of the GRC and GRS meetings provides a highly interactive and stimulating venue for cross- fertilization of ideas and development of new collaborations. The Visual System Development GRC has established a reputation as the leading conference in its field, and it is the only meeting on the topic that brings together vision researchers working on the full range of experimental systems in the field, ranging from Drosophila to human. The current proposal requests funds to help defray conference fees for attendees at both meetings. The Visual System GRC and GRS will feature scientists at the cutting edge of the field, with careful attention taken to ensure involvement of researchers around the world, with participation from scientists at all stages of their careers.

NEI - National Eye Institute

SUMMARY Rodents from the genus Acomys (Affrican Spiny mice) have recently emerged as a powerful model for regeneration, being capable of fully regenerating sections of skin without scarring including the development of new skin appendages. The regenerative capabilities of the Acomys species is not limited to the skin, with studies also demonstrating they can regenerate skeletal muscle tissue, cartilage, kidney, peripheral nerves and digits. Thus, the Acomys are a powerful animal model to understand the cellular and molecular mechanisms that promote regeneration with limited scarring in mammals. Aim 1a of this proposal will establish whether the Acomys cahirinus are capable of scarless corneal wound healing by comparing the regenerative capabilities of the cornea of A. cahirinus to the Mus musculus following alkali burn (AB). Aim 1b will then characterize and compare the mechanisms of wound healing between the A. cahirinus and M. musculus. This aim will establish for the first time whether A. cahirinus are capable of scarless corneal wound healing and identify the mechanism by which A. cahirinus are capable of regenerating a transparent cornea following AB. Our lab has been using the AB model on mice of diverse genetic backgrounds for well over a decade. Over the years, we have collated significant longitudinal data to show that when a group of wild-type mice on the same genetic background, such as C57BL/6J, are subjected to AB of equal severity, some mice are able to regenerate their corneas with limited to no scarring by day 14 (~18% of mice), while the others present corneal scarring (~82%). Curiously, our unpublished data show that the mice that present corneal scarring by day 14 present significant inflammatory cell infiltration and severe epithelial defect at day 1, while mice that are able to regenerate a transparent cornea by day 14 do not. Thus, excessive inflammatory infiltration within the first 24 hours following injury dictates whether the cornea will regenerate or suffer corneal scarring. Aim 2 of this proposal will characterize and compare the wound healing process between inbred C57BL/6J mice that are able to regenerate the cornea following AB to those that present corneal scarring, identifying for the first time key factors that direct corneal wound healing into wound resolution and regeneration instead of corneal scarring. Clinical Significance: Corneal scarring after trauma is a leading cause of vision loss in our society. To date, there are limited treatment options for preventing and treating corneal scarring, culminating in an urgent medical need for new therapeutics that can promote scarless wound healing. This proposal will identify key cellular and molecular mechanisms that regulate scarless wound healing establishing the groundwork for developing novel therapies for triggering scarless wound healing in the clinic.

NEI - National Eye Institute

Project Summary/ Abstract Disturbances in the curvature of the human cornea can lead to visual defects like myopia, hyperopia, astigmatism, or keratoconus. Therefore, understanding how such curved refractive surfaces develop is crucial for preventing or treating these disorders. In this research proposal, I plan to use the Drosophila corneal lens as a model for the human cornea. The fly is an attractive model organism due to its plethora of genetic tools and quick life cycle; and additionally, it about 70% of genes are conserved between humans and flies. The corneal lens is a biconvex ECM structure that focuses light onto the photoreceptor rhabdomeres. A major component of the corneal lens is the polysaccharide chitin, and my preliminary findings suggest that alterations in chitin levels affect corneal lens shape. To explore this idea further, I will use fly genetics, super-resolution and electron microscopy to study the detailed role of chitin in corneal lens morphogenesis (Aim 1; K99). The retina contains a fixed number of non-neuronal central cells which secrete the various corneal lens components and also provide support to the corneal lens. My preliminary data show that an excess of either central or lattice cells affects corneal lens morphology. I will first analyze how varying the numbers of central and lattice cells changes corneal lens architecture, and then using transcriptomics I will identify the genes expressed in these cells during the pupal stage (Aim 2; K99/R00). Central cells also secrete the pseudocone beneath the corneal lens, which is analogous to the mammalian aqueous or vitreous humor. Our previous work identified three proteins in the pseudocone that influence corneal lens shape. Additional pseudocone components will be identified using TurboID to find binding partners of one of these proteins. These as well as transporters and ion channels will be tested for corneal lens shape defects and glaucoma-like phenotypes in flies (Aim 3; R00). This work will provide mechanistic insights into corneal lens morphogenesis, which may be relevant to human corneal development and diseases. To accomplish the proposed research, I will combine my existing skills in developmental genetics, biochemistry and cell biology with new skills learned during my K99 training, including super-resolution microscopy and transcriptomics. During my transition into an independent position, I will solicit advice from my mentors Drs. Jessica Treisman, Gira Bhabha, Holger Knaut, Erika Bach and Hyung Don Ryoo. Their scientific advice on fly genetics, quantitative imaging and transcriptomics along with their experience of grantsmanship, mentoring, lab management, publishing and establishing fruitful collaborations will prove invaluable. My long- term career goal is to head a research laboratory that will investigate the genetic, biophysical, cellular, and molecular regulation of corneal lens shape determination. Although I have made significant progress toward this goal with research experience and publications, I firmly believe that the additional technical and career training proposed during the K99 mentored phase is necessary for my successful transition to independence.

NEI - National Eye Institute

Project Abstract This proposal aims to transform the accessibility of digital thematic maps for the 285 million blind and low vision (BLI) individuals worldwide. Thematic maps are essential for understanding complex data in various domains, including climate analysis, electoral processes, and emergency management. However, these maps are currently inaccessible to BLI individuals, as screen readers often fail to recognize them, and they lack customization options for low vision users. If an alternative is provided, it is a simple table lacking any geographic information. This project seeks to commercialize Audiom, an innovative, multimodal, and Large Language Model (LLM) chat-based digital map viewer. Unlike existing alternatives, Audiom is designed to be fully compliant with Web Content Accessibility Guidelines (WCAG), enabling BLI individuals to engage fully in scientific and civic domains. The project will develop Audiom from a prototype into a fully commercial platform that integrates with mainstream map tools, laying the groundwork for further research into non-visual and multimodal cartography. Audiom offers a unique visual experience for low vision users, allowing customization through speech, textures, and high contrast borders. For non-visual users, Audiom provides an auditory and tactile experience, enabling navigation through a map with arrow keys or touchscreen swipes, with pitch and speech indicating the value of features. The specific aims of this project are threefold: 1) Optimize user performance with the Audiom interface through user-centered design iterations; 2) Execute community-based comparative usability studies to evaluate Audiom's effectiveness for BLI and sighted users; and 3) Create, commercialize, and distribute a productionready Software Development Kit (SDK) for accessible digital thematic maps, facilitating easy integration with existing digital map tools. By achieving these aims, Audiom will not only make thematic maps accessible to BLI individuals, ensure compliance with the Americans with Disabilities Act (ADA) and the Rehabilitation Act for government entities, but also make mainstream the field of non-visual cartography. The commercialization of Audiom will empower BLI scientists, enhance civic participation, and improve the quality of life for BLI individuals by allowing them to use digital maps for the first time.

NEI - National Eye Institute

PROJECT SUMMARY Throughout childhood, individual neurons that make up our brain form connections, or circuits, between each other. These circuits guide the flow of activity across our brain for proper functioning. A fascinating feature of the brain is that it’s believed that both our genetics and our experiences as children can influence how neurons connect to each other. Understanding this process is critical to knowing how our childhood experiences influence brain development, and how disruptions in this process result in neurodevelopmental disabilities like Autism. Surprisingly, despite its importance, we have very little direct data on how these physical connections form during childhood development. Rather, our understanding of this process is largely based on interpretations of data that do not directly follow how neurons connect with each other. This is because until recently, these kinds of experiments were difficult if not impossible to do. However, our lab has developed technology that now allows us to investigate how circuits form in a clear and unambiguous way. With these new tools we will first ask how neurons make the correct connections during development by focusing on a well-studied circuit between the eyes and the brain. This circuit will allow us to ask how connections between the left and right eye properly form in concert together to ensure that what our eyes see gets correctly processed in the brain. In the second aim, we will ask how closing one eye, which will block activity, or visual “experiences” from that eye, disrupts this process. We believe that what we learn from studying this visual circuitry will provide clues to how neurons form circuits throughout the brain. We will conduct these experiments in the ferret because there is a lot of existing data on how this visual circuit develops in this animal that we can use to better interpret our results. Additionally, the ferret visual circuit and its brain more closely resembles the human brain relative to mice, another popular model organism in neuroscience. This will make it more likely that the lessons we learn in the ferret will also be true in humans. Overall, we believe results from our proposal will help better understand how brain connections form during childhood which may inform how much we need to worry about the environment our children grow up in. Our proposal might also help with treating neurodevelopmental disabilities by providing additional evidence for treating conditions early in life while the brain is forming new connections and is still amenable to therapies.

NEI - National Eye Institute

PROJECT SUMMARY Visual tasks, like reading print or recognizing faces, often require detailed spatial information collected under changing lighting conditions. In the retina, as light intensity varies so do the gain and kinetics of neural responses—a process called adaptation that prevents saturation and supports a consistent perception of contrast. A significant gap in knowledge is how light adaptation functions in the fovea—the central most region of retina responsible for high-acuity vision. Retinal circuit adaptation relies on signal pooling, and therefore adaptation may vary between foveal and peripheral regions that differ in convergence. Additionally, cone photoreceptors in peripheral primate retina are known to be asymmetric in their sensitivity to light increments vs decrements. Therefore, circuit adaptation may differ in ON and OFF visual pathways. The objective of this project is to use distinct primate retinal circuits—foveal vs peripheral and ON vs OFF—to determine how circuits with differing signal to noise ratios modulate gain and kinetics across light conditions. Aim 1 will determine the impact of convergence on properties of retinal circuit adaptation (gain, kinetics, noise, and time course) in the primate foveal vs peripheral midget and parasol pathways, as well as the contribution of synaptic inhibition as a potential mechanism for circuit adaptation to luminance. Aim 2 will determine how asymmetries inherited from cone photoreceptors shape the functional properties of adaptation in the primate ON vs OFF peripheral midget pathway. Given the importance of the fovea for our everyday vision, this work will bridge a gap in our knowledge of how foveal circuits adapt to changes in contrast over varying background luminance, critical for the function of high-acuity vision. These results will positively impact the pursuit towards prosthetic retinal implants that can recapitulate properties of foveal circuits by providing a template for function in diverse retinal circuits. The training plan described in this proposal is designed to enable me to develop the skills necessary to reach my career goal of an independent investigator. By following this plan with the guidance of my sponsor and co-sponsor, I will continue to learn new electrophysiology techniques to build a strong foundation in retinal circuit research. I will develop my writing, communication, teaching, mentoring, networking, and scientific outreach skills. This research will take place at the University of Wisconsin-Madison where the strong intellectual environment and availability of primate tissue from the Wisconsin National Primate Research Center will be leveraged.

NEI - National Eye Institute

PROJECT SUMMARY/ABSTRACT Vision loss caused by the death of photoreceptors is a leading cause of irreversible blindness worldwide, yet therapeutic options remain limited. For this reason, the NEI’s Retinal Disease Program has identified the development of strategies for the treatment of retinal degenerations as a core program goal. Recently, several laboratories have derived photoreceptors from stem cells, making cell-replacement therapies particularly promising. Additionally, important advances have been made into manipulations that could stimulate retinal regeneration from the retinal Müller glia. The critical barrier for the success of such therapies is to understand the factors required to direct fate decisions in progenitors towards fully differentiated cell subtypes that are also capable of properly rewiring into retinal circuits. Although key transcription factors have been identified as essential for generating retinal cell classes, the target genes required to generate each retinal cell subtype are still undetermined. TBX2 is a central transcriptional regulator of all photoreceptor subtypes and is highly conserved across vertebrates. Our main hypothesis for this proposal is that retinal progenitors express TBX2 to repress the identity of photoreceptor subtypes that are not UV cones through two different mechanisms, and our main goal is to identify these two roles. Three Specific Aims are proposed: Specific Aim 1 will test the hypothesis that tbx2a and tbx2b have subdivided functions and their roles in generating photoreceptor subtypes are different. Specific Aim 2 will identify candidate factors downstream of TBX2 important for the generation of photoreceptor subtypes. Specific aim 3 will test the hypothesis that differences in Tbx2a and Tbx2b function are caused by differences in the repression domain. At the successful completion of this project, I will have identified the subdivided functions of tbx2a and tbx2b and subsequent targets to generate and maintain photoreceptor subtypes. This proposal benefits from the experimental accessibility of the retina and our deep knowledge of retinal cell types and circuits, but our approach has the potential to impact the study of other neuronal degenerative diseases. In addition to my proposed research, I designed a holistic training plan that will help me advance toward my goal of leading an independent research lab. The work planned for the F31 award period will be valuable in terms of both research and training opportunities.

NEI - National Eye Institute

PROJECT SUMMARY / ABSTRACT Rehabilitation of central visual disorders like amblyopia and cortical visual impairment depends on synaptic plasticity, the changes in synaptic connections between neurons in the brain. A major regulator of synaptic plas- ticity is brain state - the moment-to-moment fluctuations in attention, arousal, emotions and other factors sep- arate from the actual content of experience - but brain states are generally left uncontrolled in treatment. Con- trolling brain state may be particularly important for brain stimulation therapies like repetitive transcranial mag- netic stimulation (rTMS), which mediate their effect through induction of neuroplasticity. The goal of this re- search proposal is to explore how attentional state - an experimentally tractable, well-understood, and disease- relevant brain state mechanism - regulates rTMS-induced neuroplasticity to the human visual cortex (Aim 1) and frontal eye fields (FEF, Aim 2). Changes in the steady-state visual evoked potential (ssVEP) contrast-response function following rTMS provide a high signal-to-noise neural readout of visual cortical neuroplasticity, while changes in psychophysical contrast discrimination sensitivity provides a perceptual readout of plasticity. During rTMS, subjects will orient attention to either the same or opposite retinotopic visual field to which rTMS is tar- geted, to determine how attentional state affects the propensity of rTMS to induce neuroplasticity. Powerful quantitative linking models will then be used to link rTMS-induced neural changes to perceptual changes, and to determine which neural changes most contribute to behavioral change (Aim 3). These experiments will pro- vide novel evidence that attentional state controls the neuroplasticity effects of brain stimulation. Moreover, they will help identify the cortical circuit mechanisms that are affected by rTMS and which of these mechanisms are most determinative of behavioral change following rTMS. Together this provides fundamental knowledge in hu- man visual cortical plasticity addressing NEI’s Area of Emphasis Biology and Neuroscience of Vision, and will inform the development of brain state control paradigms to augment the efficacy of rehabilitative neuromodula- tion therapies for visual disorders including hemineglect, cerebral scotoma, and amblyopia, in line with NEI’s core programs on Strabismus/Amblyopia/Visual Processing and Low Vision/Blindness Rehabilitation. In the process, the candidate will expand upon his background in in vivo synaptic plasticity and optical physiology in autism animal models to gain expertise in core methods of human neuroscience including rTMS, MRI, EEG, visual spatial attention paradigms, and computational modeling, learning from Stanford mentors who are au- thorities in these techniques (Dr. Nolan Williams, Dr. Tony Norcia, and Dr. Justin Gardner). He will take full advantage of Stanford’s vibrant intellectual environment, interacting with clinicians and researchers to bridge the gap between basic neuroscience bench and the clinic bedside. This training will allow the candidate to estab- lish a unique research niche at the interface of neuromodulation, neuroplasticity, and brain states and eventually lead a translational program to implement neuromodulation-assisted behavioral and rehabilitation therapies.

NEI - National Eye Institute

PROJECT SUMMARY/ABSTRACT Animals rely on visual information to interact with their environment. In contrast to visual perception, other visual functions benefit from a temporally and spatially blurred version of the surroundings – one that is sensitive to overall illumination (irradiance) instead of image detail. In mammals, these functions rely on intrinsically photosensitive retinal ganglion cells (ipRGCs) which sense light directly due to their expression of the photopigment melanopsin. There is evidence that the M1 type of ipRGC is required for a wide range of non-image-forming visual functions such as setting of circadian phase, control of the pupil, and even regulation of mood. These mechanisms of physiological control appear to be dependent on a measure of irradiance. Indeed, our lab has shown that M1s are exceptionally well-suited to encode environmental irradiance thanks to efficient temporal integration and population level responses that span the physiological range of light intensities. That said, previous studies of M1 ipRGCs have focused on the somatodendritic compartment. The hypothesis driving this proposal is that axonal and synaptic specializations make M1s even more effective at encoding and transmitting irradiance information than is currently appreciated. Aim 1 will explore the functional consequences of axonal melanopsin expression. M1 axons can travel long distance (>1mm) within the retina where they are subject to natural light stimulation. My preliminary data suggest that the axons can sense light and generate action potentials independently of the soma. I will explore the ways in which this axonal photosensitivity augments the output of M1s. Aim 2 will explore how synaptic properties further transform the output of M1s and dictate how they drive downstream circuits. M1s have been identified as vital for two regulatory behaviors in particular: circadian control (mediated by the suprachiasmatic nucleus; SCN) and pupillary control (mediated by the olivary pretectal nucleus; OPN). M1s control circadian phase on slow timescales (hours to days) but control the pupil on fast timescales (seconds to minutes). I will explore the features of the M1 synapse, in combination with those of the postsynaptic cells of the SCN and OPN, that enable M1s to drive these qualitatively and quantitatively disparate behaviors. Together, these experiments will inform our understanding of how ipRGCs encode and transmit photic information and will add to our basic understanding of the biophysical mechanisms underlying neuronal signaling.

NEI - National Eye Institute

PROJECT SUMMARY/ABSTRACT Our research aims to advance the understanding and diagnosis of Sjögren’s Disease, particularly its manifestation as dry eye disease (DED), which disproportionately affects women. By refining diagnostic capabilities, we intend to facilitate earlier detection and better therapeutic strategies, which could potentially improve patient outcomes and quality of life. We will utilize a comprehensive human proteome-wide programmable phage display to pinpoint autoantigen targets in tears collected at baseline from participants of the Sjögren’s International Collaborative Clinical Alliance (SICCA). By employing phage immunoprecipitation sequencing (PhIP-Seq), which integrates oligonucleotide library synthesis for encoding peptides in bacteriophages with high-throughput sequencing, to profile the entire antibody repertoire in tear samples. This technique allows for the high-resolution identification of autoantigens associated with Sjögren’s Disease, particularly focusing on distinguish Sjögren’s Disease from other causes of DED, especially female patients. We will also assess the persistence of specific autoantibodies identified at baseline in follow-up samples collected two years later from the SICCA cohort. This cohort includes over 700 participants with baseline and follow-up tear samples collected via Schirmer strips.

NEI - National Eye Institute

Project Summary Retinal ganglion cells (RGCs) send visual information from the retina to the brain via the optic nerve. Their degeneration underlies several major eye diseases affecting vision, including glaucoma, hereditary optic neuropathies, and ischemic optic neuropathies. Normally, RGCs do not regenerate; thus, RGC loss in these diseases is not reversible. One potential strategy for replenishing lost RGCs in patients is to reprogram stem cells and/or glial cells to functional RGCs. Knowledge about how RGCs are generated during embryonic development will greatly facilitate such efforts. During development, RGCs originate from naïve proliferating retinal progenitor cells (RPCs). Key transcription factors functioning at the different stages of RGC differentiation have been identified, and the mechanisms by which these transcription factors interact with enhancers to regulate target genes and promote RGC formation are being unraveled. These transcription factors likely interact with other factors to carry out their functions, but this is a much understudied area. The current proposal focuses on the factors that interact with Atoh7 to promote RGC formation. Atoh7 is a bHLH proneural transcription factor that is specifically required for RGC genesis. Atoh7 activates downstream genes by binding to E-box sequences in the target gene enhancers. However, multiple proneural bHLH transcription factors are expressed in the developing retina, and they are all capable of binding to similar if not identical E-boxes, yet only Atoh7 is capable of efficiently promoting RGC genesis. Using both ex vivo and in vivo assays, we have now shown that the differences between Atoh7 and other related bHLH transcription factors such as Neurod1 in promoting RGC formation lie within the bHLH domain. We have further narrowed down the responsible differences between Atoh7 and Neurod1 to six amino acid (a.a) residues. The locations of these six a.a. residues suggest that they likely provide interfaces for interaction with additional protein patterners, indicating a likely mechanism for Atoh7 to promote RGC differentiation. Our current proposal aims to identify these interacting proteins and characterize their functions. We will achieve our aims using proximity biotin labeling with our newly generated knock-in Atoh7 allele that expresses a fusion protein of Atoh7 and the biotin ligase BioID2. Our preliminary results demonstrate that this is a very feasible approach. Our specific Aims are: 1) To identify proteins interacting with Atoh7 in the developing retina and to characterize the specificity of the interactions. 2). To investigate the function of Atoh7 interacting proteins in RGC genesis. The proteins we will identify that are associated with Atoh7 will provide new research directions regarding how RGC specific gene regulation is achieved and how the RGC lineage is established. The findings will be significant not only for understanding the fundamental process of retinal cell differentiation but also for guiding efforts to regenerate RGCs to treat related retinal diseases.

NEI - National Eye Institute

ABSTRACT Our understanding of the genetic basis for inherited retinal disorders (IRDs) is in a fast forward mode. Yet the majority of the millions of individuals with IRDs remain untreated. Children with untreated IRDs often confront a lifetime of blindness with a host of concomitant burdens. Antisense oligonucleotides (ASOs) are a promising but incompletely realized option for critically needed IRD treatment. In this 2-year planning grant, a team with complementary expertise, including ASO drug development and pediatric retina and low vision, propose a step- wise plan to create a robust pathway to future treatment of pediatric IRDs due to ASO amenable genetic variants. With reference to a database of >3,000 well characterized children with IRDs, the team will streamline the analysis to determine genetic eligibility (Aim 1) namely identification of a suitable cryptic splicing defect. The records of the genetically eligible children will be reviewed to analyze clinical eligibility (Aim 2); evidence of rescuable photoreceptors will be the primary criterion. Children with rescuable photoreceptors are expected to have broad clinical spectra embracing a range of retinal diseases, syndromes, medical conditions, and developmental levels. Acknowledging that the realistic path to future ASO treatment is a patient-centric, N-of- 1/few approach, the team will develop procedures for considering not only scientific and medical criteria, but also feasibility and ethical criteria as they weigh clinical eligibility. Based on the lessons learned from Aim 1 and Aim 2, the team will build a versatile template for regulatory review, including the manual of procedures (Aim 3) in anticipation of future N-of-1 trials of ASO treatment of pediatric IRDs. The team’s work will clarify indications to ASO treatment of pediatric IRDs and the pathway to treatment. Early, effective ASO treatment of children with IRD has potential to accelerate the creation of the next generation management and treatment of IRDs.

NEI - National Eye Institute

PROJECT SUMMARY/ABSTRACT Our brain generates sensory perceptions by integrating externally (bottom-up) and internally (top-down) driven representations of the world. The bottom-up pathways provide information originating from the sensory periphery about the physical properties of the stimulus. The top-down pathways provide inferential or predictive information about the nature of the stimulus based on its context and on the organism’s experience. Imbalances between the bottom-up or top-down driven representations result in profound perceptual abnormalities, leading to various psychiatric disorders. However, the mechanisms that enable our brain to establish and maintain a balanced integration of bottom-up and top-down information streams during sensory processing is unknown. Specifically, the mammalian sensory cortices are composed of a diverse array of neuronal types classified by their transcriptomes. It is unclear which types receive inputs from the bottom-up, top-down, or both sensory streams. Additionally, while sensory neurons exhibit varying responses to the same stimuli depending on the context within which the stimuli are presented, it remains unclear whether this variability is influenced by how these neurons are integrated in the bottom-up and top-down pathways. Furthermore, the timeline for when bottom-up and top- down integration matures is largely unknown, as is the impact of experience on this developmental trajectory. I propose to address these questions using the mouse primary visual cortex (V1) as a model system. V1 receives visual information that ascends from the eyes through the dorsolateral geniculate nucleus of the thalamus (dLGN) and descends from higher-order visual areas, including the lateral posterior (LP) nucleus and cortical higher visual areas (HVAs). I hypothesize that V1 neurons receiving bottom-up or top-down inputs from the dLGN, LP or HVAs represent largely distinct transcriptomic categories with distinct laminar profiles and response properties to visual stimuli. Furthermore, I hypothesize that visual experience is necessary for the development of bottom- up and top-down integration in V1. To test these hypotheses, I will implement a combination of a newly developed anterograde transsynaptic tracing technique, spatial transcriptomics and in vivo two-photon imaging to carry out the following experiments. In the K99 phase, I will first determine the molecular identities of neurons receiving bottom-up or top-down inputs in the adult mouse V1. I will then characterize the response properties of adult V1 neurons in relation to the origins of inputs they receive and to their molecular identities (K99 + R00). In the R00 phase, I will determine the developmental time course and the role of visual experience in the development of bottom-up and top-down integration in V1. These experiments will allow us to provide the first mapping of the two major streams of visual information, the bottom-up and the top-down, to V1 onto molecularly defined neuronal types. We will also reveal, for the first time, the logic of neuronal responses to sensory stimuli in relation to the two major pathways that impinge on these neurons. Furthermore, we will uncover fundamental principles of the development of bottom-up and top-down integration in V1.

NEI - National Eye Institute

Project Summary Sensory experience during early postnatal life profoundly influences the development of neural circuits. The primary visual cortex (V1), as part of the neocortex, is a well-established site of developmental plasticity, where visual experience plays a key role in the proper development of its binocular circuitry. However, how vision regulates the organization of individual neurons in V1, the cell types and connections they form, and the molecules they express, remain poorly understood. I address this knowledge gap by combining single-cell transcriptomics, epigenomics, connectomics and computation to link genes, cell types, regulatory networks and circuit structure in layers 2 and 3 (L2/3) glutamatergic neurons in V1. My recent work found that L2/3 glutamatergic neurons in V1 form continuous rather than discrete cell types. These cells differentially express hundreds of molecules known to be important for neuronal cell-type identity in a graded fashion, and are spatially enriched in different sublayers of L2/3. In mice that are deprived of visual experience by dark rearing, this cell-type organization is disrupted in a specific way. Based on these findings, we hypothesis that the continuous and vision-dependent cell types are key structure underlying cortical developmental plasticity. Aim 1 of my research will determine the developmental origin of L2/3 cell-type continuum. I will generate a developmental atlas of the L2/3 neurons using three existing single-cell RNA-seq datasets from mice with and without visual deprivations. I will delineate the vision-dependent developmental programs of L2/3 neurons, and identify gene programs that are temporally-regulated, type-specific and/or vision- dependent, which will serve as candidates for follow-up experiments to examine their causal link in L2/3 wiring. Aim 2 will examine the axonal-projection patterns of L2/3 neurons in V1. These neurons send divergent projections from V1 to many higher visual areas (HVAs), and transcriptomic differences in L2/3 neurons are thought to be associated with differences in projection targets. We will use barcoded connectomics to systematically determine the relationship between L2/3 cell-type continuum and projection targeting specificity to HVAs. Aim 3, to be pursued during the R00 phase, will investigate the gene regulatory networks underlying L2/3 neurons. Using single-cell multiomics data, we have identified tens of thousands of genomic regions with graded differential accessibility along the L2/3 cell-type continuum. To understand the regulatory logic linking gene expression and genomic elements often known as enhancers, we will use both experimental and computational methods. Experimentally, we will perturb the expression levels of key transcription factors and examine changes in L2/3 cell-type organizations and projection patterns. Computationally, we will train a DNA language model to predict the graded chromatin profiles of L2/3 neurons, and use in silico perturbations to screen candidate genes. Ultimately, the two efforts could be combined into a closed-loop iteration where experiments improve the model, and the model prioritizes experiments. My work will investigate the influence of visual experience on genes, cell types and circuits in a subclass of cortical neurons, thereby providing insights into the molecular logic of wiring plasticity in the mammalian neocortex.

NEI - National Eye Institute

Glaucoma is a leading cause of blindness, with primary open-angle glaucoma (POAG) affecting over three million Americans. The Ocular Hypertension Treatment Study (OHTS) provides a unique, high-quality longitudinal dataset to investigate visual field (VF) progression and improve early detection of POAG-related deterioration. Current diagnostic methods often fail to identify early-stage disease transitions, leading to delays in treatment and missed opportunities for intervention. Additionally, conventional metrics such as mean deviation (MD) overlook localized changes critical for assessing VF loss progression. This project aims to develop and implement novel statistical methodologies to enhance the detection and characterization of POAG progression. Specifically, we propose: (1) a change-point model for MD progression that integrates longitudinal MD measures with the time to detectable change, addressing variability in VF deterioration and identifying early disease transitions; (2) spatial-temporal modeling of pointwise VF progression to improve sensitivity in detecting localized changes, accounting for spatial correlations across retinal locations; and (3) development of a user-friendly R package to disseminate these methodologies for use by researchers and clinicians. By leveraging advanced statistical models and the extensive OHTS dataset, this research will refine estimates of differential progression rates, facilitate early identification of high-risk individuals, and optimize treatment strategies. Beyond glaucoma, these methods will have broad applicability in analyzing spatial-temporal data in other medical domains.