2022 Shaffer Grants

Kun-Che Chang, PhD
University of Pittsburgh
Funded by Tania and Michael Stepanian

Project: A New Therapeutic Gene for RGC Survival and Axon Regeneration in Glaucoma

Summary: In patients with glaucoma or other optic neuropathies, damaged retinal ganglion cells (RGCs) die as their axons degenerate. Although many regulators have been reported to function in axon regeneration, regrowth of long-distance axons into the chiasm and brain remains a major challenge. Thus, discovery of new therapeutic factors remains a long-standing research goal. To obtain the synergistic effects of regulators on axon regeneration, we aim to discover new regulators that control different pathways from those already known, which we can then combine for cotreatment therapy. We propose a new pathway involving the molecule identified from prior study. This proposal will assess its role in RGC damage repair using an optic nerve crush model that could translate into therapies for glaucoma and optic neuropathies, which could improve impacted individuals’ quality of life. The results will reveal the functions of this new gene in RGC survival and axon regeneration, which impacts the field by providing a potential therapy for translational studies in patients with glaucoma and optic neuropathies.

M. Elizabeth Fini, PhD
Tufts University
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Mechanisms of Steroid-Induced Ocular Hypertension

Summary: Ocular hypertension (OH) is the major risk factor for glaucoma, the leading cause of irreversible blindness. Lowering intraocular pressure (IOP) is the only proven treatment. All forms of OH are caused by increased outflow resistance via the conventional outflow pathway comprising the trabecular meshwork (TBM). In open angle OH, pathology is intrinsic to the TBM. This research project focuses on a secondary form of open angle OH caused by treatment with pharmaceutical glucocorticoids (GCs) in the eye. We hypothesize that a protein called ELAVL1is responsible for stabilizing fibronectin mRNA in response to GCs. We will investigate this idea in cultured TBM cells by perturbing ELAVL1 gene expression and assessing the effects on fibronectin stability in the presence and absence of dexamethasone. Study results will be of high impact if our hypothesis is correct, providing the preliminary data for a full study to validate the mechanism in vivo and determine the mechanisms regulating ELAVL1. The ultimate goal is to learn whether GCs directly stimulate expression of a gene, via its GRE, that is responsible for ELAVL1 activation.

 Sidney Kuo, PhD
University of Minnesota
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Early Structural Changes to Müller Glial Cells in Glaucoma

Summary: Higher than normal pressure in the eye (intraocular pressure) is the most common risk factor for glaucoma. Despite intensive research, the mechanisms linking high intraocular pressure and eventual vision loss are not well understood. Based upon preliminary studies in our lab, we hypothesize that pressure-induced changes to the physical structure of Müller glial cells early in disease onset contributes importantly to the eventual death of the retinal ganglion cells, the neurons that transmit visual information from the retina to the brain. To investigate this, we will use high resolution microscopy to image fluorescently-labeled Müller cells and retinal ganglion cells in eyes of mice with experimentally-induced elevated intraocular pressure. By identifying early pathological events in Müller cells, our long-term goal is to provide new insight into possible ways to diagnose and treat glaucoma before retinal ganglion cells are irreversibly lost.

 Myoungsup Sim, PhD
Duke University
Funded by the Dr. Miriam Yelsky Memorial Research Grant

Project: Primary Cilia-mediated Nitric Oxide Production in Schlemm’s Canal Cells

Summary: Elevated intraocular pressure (IOP) results from dysfunction of the conventional outflow pathway, however, the glaucoma drugs targeting this pathway are very limited. Latest studies have shown a potential role of nitric oxide (NO) in lowering IOP. In this context, a NO-donating drug (latanoprostene bunod) was recently approved for the reduction of IOP in patients with ocular hypertension and glaucoma. Nevertheless, most of the NO-based drugs have failed to be approved by FDA due to some challenges related to the exogenous delivery of NO, such as uncontrolled NO release, suggesting that regulation of endogenous NO production could represent a better strategy for glaucoma treatment. NO is produced by the endothelial cells of the Schlemm’s canal (SC), a unique vascular structure component of the outflow pathway. As a result of elevated IOP, SC cells are subjected to fluid flow-induced shear stress, which has been reported to trigger NO production by SC to regulate IOP homeostasis. We seek to investigate how to regulate endogenous NO production in SC cells to improve the NO-based glaucoma therapy. Results of this study will provide an upstream target molecule that can regulate endogenous NO production and advance the current NO-based glaucoma drugs to treat glaucoma patients more safely.

 Brian Soetikno, MD, PhD
Stanford University
Funded by Bob and Birdie Feldman & Giving Tuesday contributions

Project: Visible Light OCT for Glaucoma

Summary: Glaucoma is characterized by the damage and death of retinal ganglion cells (RGC) and their retinal nerve fiber layer axons, leading to vision loss. Clinicians rely on diagnostic tests, such as visual field studies and optical coherence tomography (OCT), to diagnose and monitor glaucoma. However, these tests are currently insensitive to detecting glaucoma early and monitoring minute changes in its progression. This proposal will address this need by designing and testing a new OCT imaging device, called visible-light OCT (vis-OCT). Vis-OCT uses visible wavelengths (560 nm +/- 50 nm) to provide ultra-high resolution, three-dimensional imaging (1 to 2 microns) of the retinal layer anatomy. The ultra-high-resolution of vis-OCT enables sub-laminae to be distinguished within the inner plexiform layer (IPL), a site where RGC’s synapse to other retinal neurons. The ability to discern these connections opens the possibility of sensitively detecting RGC damage by measuring the thickness of the sub-laminae. Vis-OCT imaging can be technically challenging due to artifacts from patient eye motion. To solve this issue, we will construct a vis-OCT device with fundus tracking, enabling the acquisition of high-quality images even in the presence of eye motion. Using this novel device, we will image IPL sub-laminae and make measurements of the thicknesses in varying glaucoma stages. This study will provide insight into whether we can detect RGC damage at ultra-high resolution in the IPL. Ultimately, vis-OCT could offer a new imaging biomarker for detecting glaucoma early and monitoring its progression.

 Qing Wang, MD, PhD
Columbia University
Funded by the Dr. Henry A. Sutro Family Grant for Research

Project: Novel Tools to Identify and Target Astrocytic Subtypes to Treat Glaucoma

Summary: Vision loss in glaucoma results from damage to the retinal ganglion cells (RGCs), which are the neurons that connect the eye to the brain. RGC axons from throughout the retina converge at the optic nerve head to exit the eye. A critical insult to RGC axons occurs at the optic nerve head, where they interact extensively with another resident cell type, astrocytes. Astrocytes are thought to play both neuroprotective and neurotoxic roles in complex neurodegenerative diseases, such as glaucoma and Alzheimer’s. Our studies will provide important molecular information and tools for manipulating optic nerve head astrocytes to understand and treat glaucoma. We will use cutting edge single-nuclear RNA sequencing technology to dissect out the molecular pathways that define neurotoxic versus neuroprotective astrocytes in optic nerve heads of glaucoma and normal mouse eyes. We will also develop new viral tools for selectively manipulating genes in optic nerve head astrocytes that can be used for gene therapy approaches by the glaucoma research community. These studies will pave the path for the development of a new avenue of glaucoma therapies directed towards optic nerve head astrocytes.

2021 Shaffer Grants

Ta Chen Chang, MD
Bascom Palmer Eye Institute
Funded by the Harvey DuBiner, MD Memorial Fund

Project: Genetic Studies of Open Angle Glaucoma in Haitian Community

Summary: The objectives of this proposal are to identify the genes associated with primary open angle glaucoma (POAG) affecting the Haitian community by screening high-risk individuals for POAG. Glaucoma is the leading cause of irreversible blindness worldwide, with early detection/management being the best way to preventing blindness. POAG disproportionally affects Haitians individuals, with an earlier age of diagnosis and more severe disease on presentation compare to individuals of other races. The reason behind this disparity is unknown. POAG is highly heritable with several known genes, none of which has been well studied in the Haitian population. We hypothesize that the increased risk and aggressive course of POAG in Haitians may be due to one or several POAG genes being overrepresented in the population due to Haiti’s relative geographic and cultural isolation. Collectively, the work proposed is expected to establish the first high quality glaucoma genetic database of the Haitian population. This rich database will increase the diversity of glaucoma genetic study populations and lay the key steps to the personalized approach of glaucoma care (including gene-based screening strategies and therapeutics) for the Haitian community.

Qi N. Cui, MD, PhD
Stellar-Chance Laboratories, University of Pennsylvania
Funded by Dr. James and Elizabeth Wise

Project: Evaluating the Glucagon-like Peptide 1 Receptor (GLP-1R) as a Therapeutic Target in Glaucoma

Summary: Glaucoma is characterized by death of retinal ganglion cells. For patients with glaucoma, intraocular pressure reduction is the only therapeutic mechanism available to slow disease progression. Unfortunately, successful pressure lowering does not prevent disease progression in a significant number of patients. Recent studies have implicated neuroinflammation in the pathogenesis of glaucoma. Evidence from our lab and others suggest that pro-inflammatory microglia and macrophages respond to ocular hypertension by inducing astrogliosis to cause retinal ganglion cell loss. Furthermore, once present, retinal inflammation persists beyond intraocular pressure normalization. NLY01, an agonist for the glucagon-like peptide 1 receptor (GLP-1R), effectively prevents astrogliosis in ocular hypertension to rescue retinal ganglion cells. GLP-1R class of therapeutic agonists is efficacious and safe in the long-term treatment of type 2 diabetes. We hypothesize a multi-hit hypothesis of glaucoma pathogenesis, whereby RGC-autonomous mechanism of pressure-induced cell stress acts in concert with non-cell autonomous mechanism of retinal inflammation to trigger neuronal cell death. We will test this hypothesis using three models of glaucoma. In parallel, we will examine whether NLY01, and more broadly GLP-1R agonists, may be a viable therapeutic adjunct to existing pressure modifying therapies for glaucoma.

Luca Della Santina, PhD, PharmD
University of Houston, College of Optometry
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Excitatory – Inhibitory Balance in Glaucoma

Summary: Diagnosis of glaucoma often occurs when a significant amount of retinal ganglion cells die and vision is lost. Some early sub-clinical alterations occurring in this disease involve loss of synapses inside the retina. Synapses can either excite or depress the retinal ganglion cells and balance between these two types of synapses constitute a key factor regulating the function of ganglion cells. We have demonstrated that excitatory synapses are lost but we still don’t know how the balance between excitatory and inhibitory synapses is modified in glaucoma. Our project proposes to study both kinds of synapses in a model of glaucoma, where we can control the time at which ocular pressure is increased. We will use our novel computer program ObjectFinder to analyze millions of synapses, using deep learning technology to recognize them, and monitor their integrity over time. With this innovative approach, we aim to discover which synapses are affected first when ocular pressure increases, and where in the retina they are located, allowing researcher to design more sensitive early screening tests for glaucoma that may help physicians to diagnose this disease before significant vision is lost.

Jiun Do, MD, PhD
Shiley Eye Institute, University of California, San Diego
Funded by Richard and Carolyn Sloane

Project: Optic Nerve Relays for the Restoration of Visual Function

Summary: Optic neuropathies like glaucoma cause vision loss by irreversibly damaging the optic nerve and disrupting the connections between the eye and the brain. There are currently no therapies to repair this connection and restore vision. Therefore, there is a need for novel therapies to regenerate the optic nerve if restoring vision is to be possible. Stem cells have the potential to repair damage in the nervous system. Preliminary work has shown that stem cells placed in the injured optic nerve similarly survive and integrate with the visual system. In this proposal, I will explore how combining stem cells with gene editing can reform damaged circuits in the visual system and the degree to which stem cells connect to the brain. To people with glaucoma, this project is significant because it has the potential to regenerate the connections between the eye and the brain that are permanently lost due to glaucoma. The ability to reform these connections is necessary to restore vision. Therefore, this project could provide people with glaucoma who have lost vision with a treatment to restore vision.

John Fingert, MD, PhD, FARVO
Carver College of Medicine, University of Iowa
Funded by the Dr. Henry A. Sutro Family Grant for Research

Project: Single Cell Transcriptome Analysis of Glaucoma

Summary: Glaucoma is one of the three most heritable human diseases, indicating that genes are very important in its pathogenesis. We previously discovered that mutations in two genes, myocilin (MYOC) or TANK binding kinase 1 (TBK1), cause human glaucoma and we engineered models with analogous mutations in these genes and showed they also develop glaucoma. Mutations in the TBK1 gene are one of the most common, known-molecular causes of human glaucoma, however, the mechanism by which a mutation in this gene causes nerve damage and blindness is not yet known. One way to investigate the biological processes that lead to glaucoma is to examine which genes are activated as the disease develops and progresses. In this application we propose to identify the pattern of gene activation that occurs eyes that develop glaucoma due to a TBK1 gene mutation. We will use a powerful new method, single cell RNA sequencing, to analyze retinal tissue. Our experiments will show which genes are activated in key cells/tissues of the eye at various times during the development of glaucoma. This data will provide a description of what goes wrong in the eyes of patients that have glaucoma at the finest molecular level. We will use these data as roadmap to describe the checkpoints of glaucoma development and to identify opportunities to prevent or halt progression of the damage to the eye that causes vision loss in glaucoma.

Jason Meyer, PhD
Indiana University School of Medicine
Funded by Bob and Birdie Feldman and Giving Tuesday contributions

Project: Complement Pathway-mediated Neurotoxicity of Reactive Astrocytes in a Stem Cell Model of Glaucoma

Summary: Glaucoma is a common cause of vision loss characterized by the progressive degeneration of retinal ganglion cells (RGCs), the projection neurons of the retina that convey visual information to the brain. Astrocytes are closely associated with RGCs in the nerve fiber layer of the retina and optic nerve, where they play a vital role in supporting RGC homeostasis and function. However, during the course of disease, astrocytes acquire a “reactive” state that is known to contribute to neurodegeneration. However, neither the factors responsible for nor the specific mechanisms underlying reactive astrocyte-induced neurodegeneration have been completely identified, resulting in changes to how reactive astrocytes interact with RGCs. The complement cascade has been implicated in physiological processes during brain development and homeostasis, mainly involved in the maturation of synaptic circuits. Nevertheless, in disease conditions, the rapid and uncontrolled activation of the complement pathway leads to inflammation and neurodegeneration. The activation of the complement pathway has emerged as a potent modulator of reactive gliosis and neuronal damage, with the classical complement pathway involved in neuroinflammation, yet the exact mechanisms by which complement exacerbates neurodegeneration remain unclear. Recent studies have demonstrated that reactive astrocytes lead to RGC neurodegeneration in a model of glaucoma, and the characterization of astrocyte reactivity has revealed elevated complement C3 expression in astrocytes, characteristic of classical complement pathway activation. However, as many phenotypic and functional differences exist in both astrocytes and RGCs between models and humans, it is unclear how reactive astrocyte-specific changes exert their effects in the human retina. Hence, it is hypothesized that A1 reactive astrocytes mediate RGC neurodegeneration through the activation of the C3 complement pathway. In order to test this hypothesis, human pluripotent stem cell (hPSC)-derived astrocytes and RGCs will be utilized to study mechanisms underlying the toxic effects of reactive astrocytes upon RGCs in a human cellular model of glaucoma, with a subsequent analysis of how complement pathway activation leads to neurodegeneration.

Lev Prasov, MD, PhD
Kellogg Eye Institute, University of Michigan
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Elucidating the Role of a Novel Closure Associated Gene in Eye Development and Disease

Summary: Angle closure glaucoma is a blinding condition that affects 0.5% of the world’s population. It is caused by blockage of the drainage pathways in the eye, leading to acute or chronic elevations in eye pressure and subsequent damage to the optic nerve. This can lead to swift loss of vision. Disorders affecting the development or growth of the eye can lead to susceptibility to this subtype of glaucoma. The molecular mechanisms leading to angle closure glaucoma are largely unknown, but it is likely guided by underlying eye anatomy. Understanding genetic causes is the first step towards developing more effective treatments for preventing this condition. By studying a large family with angle closure and small eye size, we have recently identified a new candidate gene and mutation for this condition in a critical protein with an unknown role in eye development. This protein functions as a transcription factor regulating the expression of multiple genes in various organ systems. We hypothesize that dysfunction in this protein leads to abnormal eye development and in turn predisposition towards angle closure glaucoma. To test this, we will first evaluate the effect of our identified familial mutation on protein function. Next, we will identify the critical cells in the eye that express this protein during development. Finally, we will screen a cohort of individuals with small eyes and angle closure glaucoma for other mutations in this gene, and follow-up any candidate mutations with our established functional tests. These studies will establish the role of this regulatory protein in eye development and disease, and pave the path to investigate a new developmental pathway that leads to angle closure glaucoma.

Teresa Puthussery, BOptom, PhD
UC Berkeley School of Optometry
Funded by Molly and David Pyott

Project: A Novel Approach to Assess Selective Ganglion Cell Vulnerability in Glaucoma

Summary: Glaucoma is a progressive blinding disease that leads to the death of ganglion cells, the nerve cells that transmit visual signals from the eye to the brain. There are many different types of ganglion cell in the human retina, each of which preferentially detects different features in the environment such as color, motion and fine spatial detail. This project will use a novel approach to determine whether specific types of ganglion cells are more prone to degeneration in human glaucoma. The results of this study will inform efforts to develop improved clinical tests for early detection and monitoring in glaucoma.

Steven Roth, MD, FARVO
College of Medicine, University of Illinois
Funded by The Dr. Miriam Yelsky Memorial Research Grant

Project: Novel Slow-release Exosome Formulations for Glaucoma

Summary: Here we propose a new strategy for glaucoma, extracellular vesicles (EVs), which possess significant neuroprotective properties, linked to hydrogels for injection into the eye for prolonged EV delivery. Our aim is to develop and optimize EVs to provide neuroprotection for RGCs. The preparations will be tested and optimized using in vitro and in vivo models. This therapy using EVs fits the goals of precision medicine. EVs derived from mesenchymal stem cells (MSCs) have neuroprotective and regenerative properties and are well suited for glaucoma treatment. We will use specific binding peptides that recognize sites on the EVs to link them to alginate hydrogels. These EV/alginate preparations will be tested in vitro. In the second aim, we will test the efficacy of the preparations in a model of glaucoma. Well-developed methods including visual function and histology, and confocal imaging in vivo and in vitro will test efficacy of these EV modifications. The rationale is that medical or surgical intraocular pressure reduction, the only clinically approved glaucoma treatment, neither prevents the main pathophysiological mechanism, retinal ganglion cell (RGC) death, nor axonal loss. Administration route, dosage, and adverse effects limit clinical application of neuroprotective agents. Moreover, the complex pathology of glaucoma necessitates action upon multiple injury mechanisms, including oxidative stress, impaired axonal transport, neuro-inflammation, and excitotoxicity. Stem cells release EVs, nanoparticles that facilitate cell-to-cell communication. MSCs-EVs decrease neuronal cell death after hypox-ia/ischemia in vitro and in vivo, stimulate axonal growth, and attenuate inflammation and oxidative stress. They can be administered cross-species, and their stability, biocompatibility, biological barrier permeability, and low toxicity make them attractive therapeutic delivery vehicles. EVs are taken up by cells; unlike stem cells, which rely upon integration into tissues, or diffusion of secreted contents to the cells, EVs deliver their cargo directly. This study is significant because it will provide the first steps toward a precision therapy for glaucoma by advantageously harnessing properties of EVs and hydrogels, combined together for the first time.

2020 Shaffer Grants

Steven Bassnett, PhD
Washington University School of Medicine
Funded by The Dr. Miriam Yelsky Memorial Research Grant

Project: Role of LOXL1 Propeptide Aggregation in Pseudoexfoliation Glaucoma

Summary: Pseudoexfoliation (PEX) syndrome is the most common identifiable cause of glaucoma, afflicting millions of people worldwide. In patients with PEX syndrome, white, powdery material accumulates on the front surface of the eye lens. Under the microscope, the powdery aggregates can be seen to consist of tangled fibrils. Contraction of the pupil dislodges the PEX material and abrades the inner lining of the iris, causing release of pigment. The combination of PEX material and pigment granules can block the drainage pathways of the eye. As a result, many PEX patients experience unusually high pressure within the eye and nearly half go on to develop PEX glaucoma, a challenging condition to treat clinically because it is resistant to medical therapy. Genetic studies have suggested that inherited variations in a gene called LOXL1 are closely associated with the risk of developing PEX syndrome, but the precise link between the presence of genetic variants and the disease mechanism remains obscure. We will test the hypothesis that aggregation of the LOXL1 propeptide is an initiating event in PEX fibril formation. This will provide important insights into the etiology of PEX glaucoma.

Stewart Bloomfield, PhD
State University of New York College of Optometry
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Retinal Gap Junctions Form Novel Targets for Neuroprotective Therapy in Glaucoma

Summary: The use of IOP-lowering drugs, the current mainstay treatment for glaucoma, is often insufficient to prevent progressive visual loss in patients. Therefore, recent work on potential glaucoma treatments have shifted to assessment of neuroprotective strategies to promote neuronal survivability and thereby preserve visual function. Our strategy is to determine the mechanism(s) responsible for secondary cell loss in glaucoma to create novel preventive treatments. Our experimental program will study a novel mechanism for cell loss in glaucoma, so as to specify targets for innovative treatments to preserve cell health and visual function. While our project will focus on glaucoma, our results should inform potential treatments of other neurodegenerative diseases of the retina, such as retinitis pigmentosa and ischemic retinopathy, as well as other degenerative brain insults, such as stroke.

Alex Huang, MD, PhD
Doheny Eye Institute
Funded by Dr. James and Elizabeth Wise

Project: Investigating Subconjuctival Lymphatics for the Treatment of Glaucoma and Eye Disorders

Summary: In order to have a successful glaucoma surgery, intraocular fluid must both enter into and exit subconjunctival blebs. This latter phenomenon is much less understood. Thus, this proposal studies the biology of subconjunctival fluid outflow in order to develop strategies to enhance it for improved glaucoma surgical outcomes. We start by evaluating a long‐standing but unproven hypothesis that lymphatics drain the subconjunctival space and blebs. We utilize imaging methods which can isolate fluid outflow pathways from blebs for exact structural and molecular identification. Then, we develop methods to manipulate bleb outflow pathways using protein growth factors. We hypothesize that growing more lymphatics into subconjunctival blebs can help glaucoma surgeries akin to building rivers past a dam to move water past the obstruction. With these tools in hand we hope to better understand glaucoma surgeries, develop tools to improve them, and in so doing preserve vision with less patient reliance upon daily glaucoma drops.

Tatjana Jakobs, MD
Schepens Eye Research Institute
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: The Transcription Factor Runx1 as a Novel Mediator of Astrocyte Reactivity in the Optic Nerve

Summary: In glaucoma, the retinal ganglion cells degenerate and die, which causes the characteristic vision defects in this disease. At the moment, lowering the intraocular pressure is the therapy of choice but this is not effective in all cases. A neuroprotective approach that directly prevents ganglion cell death and that could be combined with pressure-lowering drugs would be welcome. The retina and optic nerve contain supporting cells (astrocytes) that normally carry out important tasks to help ganglion cell function. In case of injury, such as an elevation of intraocular pressure, astrocytes become reactive. This is at least initially a protective response that aims to save retinal ganglion cells and their axons from degeneration. We have studied the molecular mechanisms of astrocyte reactivity and identified a transcription factor (Runx1) that is up-regulated in reactive astrocytes. Our hypothesis is that Runx1 directs the expression of other genes in the astrocytes that support ganglion cell survival. We want to identify these genes and the proteins they encode. Ultimately, the goal is to provide neuroprotective factors therapeutically in glaucoma.

Rachel Kuchtey, MD, PhD
Vanderbilt Eye Institute

Project: Investigation of Ocular Biomechanical Defects in Mice with Microfibril and Elastic Fiber Defects

Summary: Age-related changes in the stiffness of ocular tissue have been well recognized in glaucoma, though how these alterations take place is not fully understood. Two challenges are lack of good models and sophisticated tools to measure tissue mechanical properties. We and others have reported causative genetic mutations in ADAMTS10 and ADAMTS17 in dogs with inherited glaucoma. Because both genes encode microfibril-associated proteins, we have focused on well-characterized and well-established mouse models with microfibril defects caused by Fbn1 mutations. Fbn1 mutant mice have multi-organ biomechanical defects, and we recently reported ocular findings related to glaucoma in those mice. We believe those mice could be a good model to study ocular biomechanical properties. We further hypothesize that disruption of an additional key component of elastic fibers would result in more severe ocular biomechanical defects. We will use Atomic Force Microscopy to address the other challenge, lack of tools for measuring small tissues, such as mouse aqueous outflow and optic nerve.

Herbert Lachman, MD
Albert Einstein College of Medicine
Funded by the Edward Joseph Daly Foundation

Project: Gene Expression Profiling in Trabecular Meshwork Cells derived from Induced Pluripotent Stem Cells made from Patients with Lowe Syndrome, a Genetic Disorder that causes Cataracts and Glaucoma

Summary: Glaucoma is one of the most common causes of blindness. Although researchers have been studying its underlying biological basis for decades, blindness still occurs far too often. New scientific breakthroughs have provided unique opportunities to study the disorder from a fresh perspective, with the ultimate goal of discovering novel therapies. One breakthrough is the development of induced pluripotent stem cell (iPSC) technology, which allows investigators to turn white blood cells or skin cells into virtually any other cell type in the body – including eye tissue. Another breakthrough is the ability to analyze the expression pattern of every gene in a cell, which provides a window into the function of that cell, and how that function goes awry in disease. In this proposal, we plan on generating eye tissue from iPSCs made from individuals with Lowe Syndrome, a rare genetic disorder that leads to congenital cataracts and glaucoma. We will study the gene expression pattern in eye tissue derived from the iPSCs to find novel pathways involved in the development of glaucoma.

Matthew B. Veldman, PhD
Medical College of Wisconsin
Funded through a special gift from Akorn, Inc. and their employees

Project: Zebrafish Retinal Ganglion Cell Survival in the Context of Pro-Apoptotic Bax Signaling

Summary: In glaucoma, elevated internal eye pressure or other events such as inflammation cause damage to the nerve sending signals from the eye to the brain. Following this injury, the cells that directly connect the eye to the brain die resulting in loss of vision. This is also true in non-human, mammalian models of the disease, however in animals such as fish the injured cells survive, and vision recovers. How these animals recover after nerve injury is not well understood, but the genes and proteins involved are highly evolutionarily conserved with mammals and people. Therefore, it is likely that mechanisms supporting cell survival and regeneration in fish will be translatable to glaucoma patients and might suggest therapeutic targets. Cell death in glaucoma models and likely patients is dependent upon the protein BAX which is part of the programed cell death pathway. This gene and pathway are conserved in fish, yet injured cells do not die following injury. The goal of this project is to establish a system for studying this gene and pathway in the eye of the zebrafish model organism and determine how it remains turned off following nerve injury.

Trent A. Watkins, PhD
Baylor College of Medicine

Project: Highly Parallel Assessment of RGC Regenerative and Neuroprotective Targets

Summary: The loss of vision in glaucoma is a consequence of the disconnection of the nerve fibers that connect neurons in the eye with neurons in the brain. Our lab is developing a system to substantially increase our capacity to evaluate potential therapeutic interventions to preserve and repair these connections. Our proposed system tackles, in parallel, three interrelated biological processes that contribute to irreversible vision loss in glaucoma: (1) the degeneration, or “die-back,” of optic nerve fibers, such that they are no longer available to deliver visual information to the brain; (2) the failure of these fibers, unlike those of peripheral nerves, to regenerate and re-establish functional communication; and (3) the tendency of diseased retinal neurons to commit cellular suicide, precluding any hope for later repair. Current evidence suggests that successfully enable preservation and restoration of vision will require a combination of interventions that influence each of these processes. We propose that utilizing the remarkable sensitivity of molecular barcoding will allow for testing multiple candidates in parallel and in combination for their impacts on preserving and restoring connections between the retina and the brain.

2019 Shaffer Grants

Steven Barnes, PhD
Doheny Eye Institute
Funded by Roberta and Robert H. Feldman

Project: Functional Resilience of Retinal Ganglion Cells During Mitochondrial Dysfunction

View and download Dr. Barnes’s final project research poster.

Summary: The production of energy in retinal ganglion cells is accompanied by metabolic byproducts, many of which are damaging to cellular function. The issue of how these byproducts modulate the excitability of retinal ganglion cells (RGCs) bears heavily on the development, impact, and early detection of optic neuropathies, including glaucoma. Our goals in undertaking this project were to test novel hypotheses about how the metabolic milieu of RGCs affects their electrical signaling of visual information. Our investigations identified the characteristics and biophysical origins of changes to the physiological properties of RGCs due to oxidizing byproducts in the retina. This new knowledge will increase understanding of both normal retinal physiology as well as the pathophysiology of glaucoma. Our novel observations support an emerging model of early stages of glaucoma where increases in oxidizing chemical species during energy production, but not necessarily bioenergetic failure, leads to preferential degeneration of certain subtypes of retinal ganglion cells, resulting in loss of specific aspects of visual capabilities.

Adnan Dibas, PhD
North Texas Eye Research Institute
Funded by the Edward Joseph Daly Foundation

Project: Endothelin Converting Enzyme Knockdown is Neuroprotective in Glaucomatous Neuropathy

Summary: Very little is known about how and why the optic nerve is progressively damaged in glaucoma. Conditions known to cause glaucoma such as elevated intraocular pressure (IOP), hypoxia, and ischemia were found to be associated with changes in the expression of aquaporin water channels in non-ocular tissues (e.g. brain, neurons, and muscle). Conditions known to cause glaucoma such as elevated IOP are associated with increased small proteins known as endothelins. Data obtained in the current study suggests that elevation of IOP in rats resulted in the activation of endothelin-producing enzymes known as ECE. Therefore, reduction of ECE activities may be a novel therapeutic in the treatment of glaucoma. Currently glaucoma medication involves only pressure lowering medication; however, vision loss continues. Therefore, the identification of additional mechanisms that continue to promote vision loss will assist in the development of combination therapy of lowering pressure and preventing vision loss in glaucoma.

Daniel M. Lipinski, PhD
Medical College of Wisconsin
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Development of rAAV Vector Technologies to Facilitate Topical Gene Delivery to the Cornea

View and download Dr. Lipinski’s final project research poster.

Summary: New genetic material can be delivered to cells of the cornea via injection of a viral vector into in to the front chamber of the eye. Whilst this method is largely effective, it substantially increases the risk of patients developing complications, including eye infections, such as endophthalmitis or uveitis. Furthermore, injecting extra fluid into the eye leads to a transient, but dramatic, spike in intraocular pressure that may potentially accelerate vision loss in patients with glaucoma. In this project we explored an alternative gene delivery approach wherein the virus vector was attached to the inside of a contact lens, rather than being injected, and then the contact lens placed on the front of the eye for a brief period (30 minutes). Our main findings were that virus vectors immobilized on to a contact lens remain active, they stay in close contact with the cornea for long periods of time, and are able to penetrate the cornea and successfully deliver new genetic material effectively. These findings represent a positive step towards the development of a non-invasive gene therapy treatment for glaucoma.

Biraj Mahato, PhD
University of North Texas Health Science Center
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Chemically Reprogrammed Retinal Ganglion Cell Therapy to Treat Glaucomatous Neuropathy

View and download Dr. Mahato’s final project research poster.

Summary: Retinal ganglion cell (RGC) death is the hallmark for glaucomatous damage leading to irreversible vision loss and no curative treatments are available. RGC replacement therapy has the advantage of being applied in patients to restore vision. We have identified a set of five chemicals (5C) that can convert fibroblasts into chemically induced RGC-like cells (CiRGCs). These CiRGCs express cardinal genes and function in dish similar to their native counterparts. When transplanted into a rodent glaucoma model, CiRGCs can restore retinal function. CiRGC carries extraordinary translational potential and offer hope for patients with severe, advanced glaucoma who do not have enough residual functional RGCs to be a suitable candidate for traditional gene therapy.

Pierre Mattar, MSc, PhD
Ottawa Hospital Research Institute
Funded by Carolyn and Richard Sloane

Project: Programming and Reprogramming for Retinal Ganglion Cell Replacement Therapy

Summary: Although glaucoma is frequently treatable with medication or surgery, a large proportion of afflicted individuals are diagnosed too late to prevent the death of key retinal neurons, which are called retinal ganglion cells (RGCs). As RGCs are the only neurons that transmit information from the eye to the brain, their loss results in permanent vision impairment. Although RGCs cannot be naturally regenerated, they can be produced artificially from retinal stem cell cultures. Stem cell cultures potentially offer the promise of recovery for vision loss. However, in order to bring this approach to the clinic, we will need to greatly improve our ability to produce these cells efficiently. We have been developing an approach that will allow RGC production to be enhanced. We have identified several genetic modifications that enhance RGC production, using genes that have been shown to perform this function during the natural development of the eye. Applying these genetic modifications to retinal stem cell models is expected to facilitate the production and transplantation of RGCs for glaucoma therapy.

Lauren Katie Wareham, PhD
Vanderbilt University Medical Center
Funded by Dr. James and Elizabeth Wise

Project: Investigating the Role of the NO-GC-1-cGMP Signaling Pathway in Glaucoma

Summary: The GC1 murine model of glaucoma aligns well with the pathophysiology seen in primary open angle glaucoma patients; the disease progresses with age, and mice have moderately elevated IOP, leading to degeneration of the optic nerve. This mouse model is novel, and the work completed in this proposal has already highlighted significant transcriptional changes that may underlie the pathology observed in these mice, and thus may eventually translate to human subjects. In the short term, investigating further the role of cGMP in glaucoma will highlight pertinent pathways in glaucoma pathology that may be targeted with pharmaceuticals to delay progression of the disease. We have already established that an FDA-approved drug, tadalafil, prevents neurodegeneration in two mouse models of glaucoma. We now aim to investigate the mechanisms behind this finding in order to highlight additional downstream drug targets. We are attempting to undertake the first step in this vision; by augmenting inflammatory pathways with drugs in murine chow, we aim to investigate whether cGMP neuroprotection is dependent on inflammatory modulation, and whether this leads to increased degeneration/neuroprotection of retinal ganglion cells. In the long term, this work furthers the exciting finding that modulating cGMP signaling increases retinal ganglion cell survival. The RNA sequencing analysis will provide a wealth of data, which will be published and an open resource for mining, which may highlight other relevant gene targets for future investigation by us and other groups.

Pete A. Williams, PhD
Karolinska Institutet
Funded by the Dr. Henry A. Sutro Family Grant for Research

Project: Targeting Neuronal Mitochondria for Neuroprotection in Glaucoma

View and download Dr. Williams’s final project research poster.

Summary: Current glaucoma treatment strategies only target IOP, the principal treatable risk factor. Neuroprotective treatments for glaucoma are of great therapeutic need. We have recently discovered a role for nicotinamide (a form of vitamin B3) in protecting the optic nerve in glaucoma by targeting these metabolic processes that change in the retina early during glaucoma. Nicotinamide is well tolerated, inexpensive, and robustly protective and thus may be an ideal therapeutic candidate for human glaucoma patients. To this, recent evidence demonstrate that nicotinamide is low in the sera of primary open angle glaucoma patients and we have been part of a multi-national collaborative group that has demonstrated that oral nicotinamide can increase visual function in existing glaucoma patients. Given nicotinamide’s obvious clinical utility, more research is warranted on understand that exact roles of nicotinamide in retinal ganglion cell and optic nerve health in aging and disease. In this project we have demonstrated that mitochondria rapidly change in morphology during retinal ganglion cell injury at time points prior to gross cell death. In both mouse and rat systems that mimic glaucoma-related insults we report that nicotinamide prevents these early mitochondrial changes and protects from retinal ganglion cell neurodegeneration, extending our knowledge about nicotinamide’s role in glaucoma.

Dr. Williams was awarded the 2021 Shaffer Prize for Innovative Glaucoma Research. The Shaffer Prize, presented annually by Glaucoma Research Foundation, recognizes a researcher whose project best exemplifies the pursuit of innovative ideas in the quest to better understand glaucoma.

2018 Shaffer Grants

Monica M. Jablonski, PhD
University of Tennessee Health Science Center
Funded by the Edward Joseph Daly Foundation

Project: Extended Release IOP-Lowering Formulation

View and download Dr. Jablonski’s final project research poster.

Summary: Glaucoma is the leading cause of irreversible blindness in the world, which is projected to affect about 6.3 million Americans by 2050, and intraocular pressure (IOP) is a leading contributor to glaucoma. Our project addresses the major limitations of current glaucoma therapy by advancing a sustained-release IOP-lowering formulation that coordinates multiple targets to normalize IOP. Minimum performance metrics for the novel formulation will position us to move toward clinical trials and eventually to commercialization.

Mary J. Kelley, PhD
Oregon Health & Sciences University
Funded by Dr. James and Elizabeth Wise

Project: Trabecular Meshwork Stem Cells and the Identification of the Laser Factor

View and download Dr. Kelley’s final project research poster.

Summary: A healthy normally functioning eye is dependent upon a certain number of cells for good vision. The primary risk factor for glaucoma is elevated pressure inside the eye (intraocular pressure). The trabecular meshwork or TM is responsible for controlling this pressure, but with age and glaucoma the TM loses cells and sometimes can no longer perform its regulatory function. Glaucoma can be treated for some limited amount of time with a laser, which will burn holes in the TM, and restore the pressure to normal. Previously, we found that when human cadaver eyes are treated with a laser, there is increased cell division and migration to the areas burned. These laser-treated eyes were put into organ culture with growth medium, and when this medium was later collected, it now had molecules from the cells of the laser-treated eyes and is called “conditioned media”. When this conditioned medium was placed on non-laser treated human cadaver eyes, cell division and migration also increased in these untreated eyes, suggesting that the growth medium contained secreted substances from the laser treated TM cells. Laser-treated cells of the first eye produced a “laser factor” into the medium which increased TM cell division and migration. This project involves the isolation and identification of this factor. If we can identify what this substance is, it can be chemically synthesized and added to eye drops for the patient. Treatment with such eye drops could stimulate the stem cells of the TM in glaucomatous eyes to increase cell division and migrate to the areas needed to restore function to the eye. Our results indicate that a growth factor, PDGFbb, is a molecule important in cell division, and caused the largest amount of increase in cells. However, TNF-alpha and IL-1 seem promising as well, and there may be other factors that are important to this process. Further studies are ongoing on cell migration and on affirming our initial results with cell division.

David Krizaj, PhD
University of Utah
Funded by the Dr. Miriam Yelsky Memorial Research Grant

Project: Regulation of Tensile Homeostasis in the Trabecular Meshwork

View and download Dr. Krizaj’s final project research poster.

Summary: Given that pressure-lowering eye drops are by far the most common treatment for glaucoma it is astounding that we know so little about how the effects of pressure are sensed and transduced in the eye. The project supported by GRF identified and studied new classes of ion channels that are specialized for pressure transduction. Our studies showed why and how pressure is such a potent regulator of ocular physiology and also identified pressure-sensing ion channels as targets for a novel generation of anti-glaucoma drugs. The take-home message is that trabecular meshwork cells, which play the central role in intraocular pressure regulation, exist in the state of “tensile balance” that is maintained by dynamic activation of at least three different pressure sensitive channels. Our results suggest that the loss of pressure regulation in glaucoma may occur in part due to the disturbed balance between these pressure sensors.

Yvonne Ou, MD
University of California, San Francisco
Funded by Roberta and Robert H. Feldman

Project: Ganglion Cell Dysfunction in Glaucoma

View and download Dr. Ou’s final project research poster.

Summary: Glaucoma is an irreversible blinding disease in which the cells that comprise the optic nerve, the retinal ganglion cells (RGCs), are damaged and die. A major gap in taking care of glaucoma patients is that we do not have an objective test that measures how well the RGCs are functioning. There are actually over 30 types of RGCs, and our laboratory has recently identified specific types of RGCs that are more vulnerable in glaucoma. Taking advantage of this knowledge, we are developing novel methods to assess the function or health of RGCs that are more vulnerable versus more resistant to damage. A more sensitive and objective test of RGC function and health will greatly improve our ability to take care of glaucoma patients and their vision.

Padmanabhan Pattabiraman, PhD
Case Western Reserve University
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Anti-fibrogenic Matricellular Protein CCN1 as a Novel Therapeutic Target to Lower Intraocular Pressure

Summary: Critical barriers in the development of efficient IOP lowering therapies could be overcome if there were a better mechanistic understanding governing ECM homeostasis, TM stiffness and the pathobiological basis of altered ECM deposition in the aqueous humor outflow pathway leading to increased stiffness and outflow resistance. Our research shows that targeting CCN1 can be beneficial in increasing aqueous humor outflow. The data we find indicates that increasing CCN1 expression is able to attenuate the TGFβ2-mediated increase in TM stiffness by lowering pro-fibrogenic activity in the HTM cells caused by TGFβ2. We believe that since the CCN1 lowers the tissue stiffness and attenuates TGFβ2-mediated effects on HTM cells, CCN1 is a therapeutic target to lower IOP.

Giuliano Scarcelli, PhD
University of Maryland
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Noncontact Mechanical Mapping of the Optical Nerve Head with Brillouin Microscopy

View and download Dr. Scarcelli’s final project research poster.

Summary: The mechanism by which elevated IOP levels induce degradation of the optical nerve head (ONH), the initial site of injury in glaucoma, is not yet understood but it is widely suspected that a main culprit is the lack of mechanical balance between IOP-induced strains and the stiffness of ONH and surrounding sclera. However, we do not have viable technology to test the stiffness of the back of the eye. We have developed a potential solution to this need, an all-optical approach to mechanical measurements using Brillouin light scattering. Our technology is already in human clinical trials for transparent ocular tissues (cornea, lens). In this grant we extended the reach of Brillouin microscopy to the back of the eye by using Adaptive Optics (AO). We have demonstrated that our new AO-Brillouin microscope enables imaging opaque tissues such as ONH and sclera, we have validated the instrument to show we can measure the expected mechanical differences at the back of the eye. Thus, this pilot grant provided proof-of-principle for the use of this new technology in glaucoma animal models to measure novel functional/mechanical parameters of the sclera and ONH.

Dorota Skowronska-Krawczyk, PhD
University of California, San Diego
Funded by the R. David Sudarsky Charitable Testamentary Trust

Project: Eliminate to Protect

View and download Dr. Skowronska-Krawczyk’s final project research poster.

Summary: Glaucoma is a group of optic neuropathies characterized by slow, progressive loss of retinal ganglion cells (RGCs), degeneration of the optic nerve and, consequently, loss of vision. Although the main risk factors associated with the development of the disease are elevated intraocular pressure and aging, genetic studies have described a number of loci in the genome that further increase the risk of glaucoma. Our previous work has indicated that upon intraocular pressure elevation RGCs become senescent and affect surrounding cells. In our project, we proposed to remove early senescent RGCs in the glaucomatous eye as a way to protect neighboring RGCs from death. Using mouse model of hypertension in the eye we were able to show that early removal of affected-senescent cells is beneficial for the eye in two ways: i) Fewer ganglion cells are lost in the retina; ii) Remaining cells in the retina are functional and overall visual functions are preserved. Therefore, our project provided a solid foundation for future studies on potential applications of senolytic drugs in glaucoma patients as a way to prevent the glaucoma progression.

Dr. Dorota Skowronska-Krawczyk was awarded the 2020 Shaffer Prize for Innovative Glaucoma Research. The Shaffer Prize, presented annually by Glaucoma Research Foundation, recognizes a researcher whose project best exemplifies the pursuit of innovative ideas in the quest to better understand glaucoma.

Trent A. Watkins, PhD
Baylor College of Medicine
Funded by the Dr. Henry A. Sutro Family Grant for Research

Project: Elucidating the Dynamics of the Neuronal Stress Response in Driving the Death of Retinal Ganglion Cells

View and download Dr. Watkins’s final project research poster.

Summary: In this study, we have developed a new tool for understanding how the efforts of retinal neurons to repair themselves in glaucoma can ultimately contribute to their demise. Previous research had revealed that one potential means by which vision is lost is through a series of events that results in neuronal suicide. Retinal neurons essential for vision are challenged by disease-related changes, and their efforts to adapt to these challenges include cellular signaling that promotes repair but can also lead to cell death. We have established a tool that allows us to stimulate this signaling at various intensities and patterns to probe how and when it goes from beneficial to detrimental. Our findings so far suggest that modest stimulation may, when combined with other interventions, improve neuronal repair and survival.

2017 Shaffer Grants

Adriana Di Polo, PhD
University of Montreal
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Regeneration of Retinal Ganglion Cell Dendrites: Stimulating Connections to Restore Vision in Glaucoma

View and download Dr. Di Pilo’s final project research poster.

Summary: Loss of vision in glaucoma results from the irreversible death of retinal ganglion cells (RGCs). A crucial step towards circuit repair in glaucoma is to promote damaged RGCs to regenerate not only axons, but also dendrites to successfully reconnect with their synaptic partners. In this study, we tested the hypothesis that insulin will stimulate dendrite regeneration and the reestablishment of synaptic connections thus improving survival and function in injured RGCs. Using a range of genetic, pharmacological, imaging, and electrophysiological in vivo approaches, we show that insulin promotes striking RGC dendrite and synapse regeneration in injured RGCs. Importantly, insulin promoted robust neuronal survival and rescued light-triggered retinal responses.

Our study reveals that adult RGCs are endowed with the ability to effectively regenerate dendrites and synapses once they have been lost, and identifies insulin as a powerful strategy to restore dendritic morphology and enhance the function and survival of these neurons. Collectively, our data support the rationale for using insulin and its analogues as proregenerative therapeutic targets to counter progressive RGC neurodegeneration and vision loss in glaucoma. In summary, our findings are innovative and a major scientific advancement in the field.

This work was accepted for publication in the prestigious journal Brain (July 2018 issue).

Dr. Adriana Di Polo was awarded the 2019 Shaffer Prize for Innovative Glaucoma Research. The Shaffer Prize, presented annually by Glaucoma Research Foundation, recognizes a researcher whose project best exemplifies the pursuit of innovative ideas in the quest to better understand glaucoma.

John G. Flanagan, OD, PhD
University of California Berkeley
Funded by Dr. James and Elizabeth Wise

Project: The Role of Lipoxins in Neuroprotection: A Pathway to Understanding Glaucoma

Summary: Glaucoma is a leading cause of blindness and is associated with degeneration of nerves in the retina of the eye. We have discovered that in the normal eye small molecules called lipoxins, are released by cells that support and maintain the nerves. Under stress, as happens in glaucoma, these cells appear to stop producing enough of the neuroprotective lipoxins and the neural cells and their axons start to die. We propose to study the role of lipoxins in protecting the nerves of the eye and how they might be involved in the development of glaucoma. To do this we will use a newly developed rodent model that enables the pressure in the eye to be moderately raised over several months. We will also use specially bred mice that are unable to normally use the lipoxin molecules. This will allow us to understand the pathways and mechanisms by which lipoxins can protect the eye, and potentially develop new approaches to the treatment of glaucoma.

Brad Fortune, OD, PhD
Devers Eye Institute, Portland, OR
Funded by the Dr. Miriam Yelsky Memorial Research Grant

Project: Axonal Transport of Mitochondria: Developing an In Vivo Imaging Assay for Glaucoma Research

View and download Dr. Fortune’s final project research poster.

Summary: Despite astounding recent advances in technological capabilities that enable earlier and more accurate diagnosis of glaucoma, the fundamental events that lead to progressive axon degeneration in glaucoma remain incompletely understood. Thus, even when glaucoma is accurately diagnosed at an early stage, there is still a significant risk for progressive loss of vision, which can be severe in some cases despite successful treatment to lower intraocular pressure. A fuller understanding of the sequence of physiological events that lead to axon damage should provide new targets for both diagnostic technology and therapeutic intervention to be applied during a stage when axons are susceptible, prior to the stages of irreversible degeneration upon which current diagnostic paradigms are based. In this project, we plan to develop an assay of axon transport of mitochondria that can be applied in the living eye to study early pathological events in experimental models of glaucoma. Mitochondria are the motile power plants that supply the fundamental source of energy all along each axon for maintenance of its basic functions, most importantly, conduction of electrical signals to the brain. Evidence suggests that abnormalities of mitochondrial function and transport are among the earliest events after axon injury. There is even evidence from clinical studies suggesting that human beings with better mitochondrial function are less susceptible to glaucoma. Thus, having a reliable assay of mitochondrial transport will be beneficial for future studies to investigate the role of this critical function in the early sequence of glaucomatous axon damage.

Markus H. Kuehn, PhD
The University of Iowa
Funded by the 2017 Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: A New Look at the Role of Microglia in Glaucoma

Summary: The retina and the optic nerve are populated by microglia, a cell type supporting neurons. In glaucoma activation of these cells is known to result in the production of toxic molecules that lead to neuronal destruction. However, our preliminary data suggest that suppressing the activity of these cells may not be a beneficial therapeutic strategy. We propose that the response of microglia to glaucoma damage may have two stages. There is clear evidence that activity of microglia can induce damage in glaucoma, but we propose that this is only true in late-stage disease and that during the early stages of the disease microglia exert a protective effect. We will also determine the level of pro-inflammatory cytokines during this process.

Alan L. Robin, MD
University of Maryland School of Medicine
Funded by the Glaucoma Research Foundation Board of Directors

Project: Meducation: A Randomized Controlled Trial of an Online Educational Video Intervention to Improve Technique and Adherence to Glaucoma Eye Drops

View and download Dr. Robin’s final project research poster.

Summary: Glaucoma patients rarely report receiving instruction on eye drop technique from their doctors, and doctors have little time to instruct patients on eye drop technique. A short educational video that can be watched online at home or on mobile devices may help patients learn correct eye drop technique more easily. This study will be the first randomized trial of an educational video for improving eye drop technique. The Meducation® eye drop technique video from Polyglot Systems instructs patients on all the critical steps of proper eye drop technique in easily understandable language with animations to demonstrate each step. This project will be significant because patients who successfully learn better eye drop technique can have a greatly improved chance of performing the crucial skill of eye drop instillation correctly, without added burden to overworked clinicians. By learning better eye drop technique, patients can avoid vision loss and blindness, as well as painful medication side effects and eye infections from contaminated eye drop bottles. The video is easy to disseminate to patients nationwide and does not take any clinician time to deliver.

This work was accepted for publication in the journal Patient Education and Counseling (December 2018 issue).

Gulgun Tezel, MD
Columbia University, New York, NY
Funded by the Dr. Henry A. Sutro Family Grant for Research

Project: Autophagy in Neurodegeneration and Neuroinflammation in Glaucoma

View and download Dr. Tezel’s final project research poster.

Summary: Glaucoma is a leading cause of blindness affecting millions of Americans. However, current treatment strategies are not sufficient to prevent progressive injury to specific nerve cells and continuous loss of visual function. To better understand and treat this blinding disease, our proposed project specifically aims to determine the disease-causing importance of a specific molecular process (named autophagy) in experimental glaucoma models. For this purpose, we will model glaucoma in specific mouse strains lacking the activity of specific molecules in nerve cells or glia (another important cell type that are adjacent to nerve cells and play diverse roles to support nerve cells or contribute to inflammatory nerve injury) and analyze specific responses of neurons and glia using up-to-date analysis techniques. We expect that this project will enable us to value whether therapeutic manipulation of autophagy provides protection against nerve inflammation and injury in glaucoma. The new information should help develop new treatment possibilities for patients suffering from this disease.

Carol B. Toris, PhD
Case Western Reserve University, Cleveland, OH
Funded by The Alcon Foundation

Project: Lowering of IOP by Improved Drainage through the Ciliary Muscle

View and download Dr. Toris’s final project research poster.

Summary: Our proposed research sought to understand how movement of the muscle within the fluid drainage pathway of the eye (ciliary muscle) affects the eye pressure. This muscle has two functions; it allows us to change our focus to clearly see near or far objects, and it is a pathway for fluid drainage from the eye (uveoscleral outflow). We can change our focus at will, which means we have conscious control over this muscle. Our study supported the idea that the more we move the muscle (change our focus), the more fluid is squeezed out of the eye and the lower the eye pressure. This idea was tested in three groups of adult volunteers aged 20-25 years, 40-49 years and 60-69 years. We tested how much the eye pressure changed when staring at a distance, when focusing up close, or when alternating between near and far vision. This was done for 10 minutes per test with a 20-minute rest in between tests. The results showed that alternating accommodation lowered eye pressure significantly and surprisingly the age of the person did not make a difference. This project helped to better understand how the ciliary muscle drains eye fluid and controls eye pressure. In a future study we will investigate glaucoma patients with high pressure with the ultimate goal of finding better treatments for this blinding disease.

Tara Tovar-Vidales, MS, PhD
University of North Texas Health Science Center, Fort Worth, TX
Funded by The Alcon Foundation

Project: Role of microRNAs (miRNAs) in Pathologic Fibrosis in the Glaucomatous Optic Nerve Head

View and download Dr. Tovar-Vidales’s final project research poster.

Summary: In glaucoma, there is extracellular matrix (ECM) remodeling of the optic nerve head (ONH). The ONH astrocytes and lamina cribrosa cells synthesize ECM proteins to support the ONH. However, in glaucoma, these cells cause the detrimental changes to the ONH. We want to understand the regulation involved in glaucomatous ECM remodeling of the ONH. In this study, we examined microRNAs (miRNAs) which are small molecules that silence gene expression. In this project, we treated human astrocytes and lamina cribrosa cells with or without the profibrotic cytokine TGFβ2 to compare miRNA profiles. Our results identified profibrotic and anti-fibrotic miRNAs that are dysregulated in both astrocytes and lamina cribrosa cells with TGFβ2 treatment. We also observed by using small molecules that act as miRNA mimics, we can block the effects of TGFβ2 induced ECM expression or ECM related proteins that are associated with glaucoma. We believe miRNAs are of interest to help us understand the cellular mechanisms that occur in the glaucomatous ONH and may provide a novel therapy to treat glaucoma patients.

2016 Shaffer Grants

Kevin Park, PhD
University of Miami Miller School of Medicine, Miami, FL
Funded by The Melza M. and Frank Theodore Barr Foundation, Inc.

Project: Axon-astroglial Interaction and its Effects on Optic Nerve Repair

View and download Dr. Park’s final project research poster.

Summary: In glaucoma, the optic nerve which sends visual information from eye to brain gets damaged. Once damaged, the optic nerve does not regrow back to the brain, resulting in permanent blindness. Therefore, to restore visual function in glaucoma patients, it might be necessary to promote injured optic nerves to regenerate and reconnect to their original targets.

In the last several years, researchers have identified gene therapies that can promote optic nerve regeneration. However, there is a still major problem. Optic nerves are mostly incapable of growing straight back to the brain, and often fail to reach the brain. In our research, we seek to understand the cellular and genetic factors that prevent these nerves to correctly find their targets. Towards this goal, we first discovered that optic nerves regenerate physically on the surface of astrocytes which are the support cells in the optic nerve. Therefore, we reveal that optic nerve regeneration and navigation are in fact shaped by astrocytes.

Second, we discovered that certain genes, namely Ncad expressed in the astrocytes, are important for optic nerve interaction with astrocytes, and for optic nerve regeneration. Our study identified the key cellular and genetic players that shape optic nerve regeneration and navigation. Ultimately, our research will help elucidate factors that prevent proper optic nerve regeneration and guidance, and to find strategies that promote reconnection of damaged optic nerves and restore visual functions following optic nerve damage.

Ian Pitha, MD, PhD
Johns Hopkins University, Wilmer Eye Institute, Baltimore, MD
Funded by Dr. James and Elizabeth Wise

Project: Neuroprotection through Altered Scleral Biomechanics

View and download Dr. Pitha’s final project research poster.

Summary: To date, the only way to stop vision loss from glaucoma is intraocular pressure (IOP) reduction by daily medication use, laser procedure, or incisional surgery. In some patients IOP reduction is difficult to accomplish or glaucomatous vision loss occurs despite substantial IOP reduction.

These clinical situations highlight the need for development of IOP-independent, glaucoma treatment strategies otherwise known as neuroprotection. One promising neuroprotective therapeutic for glaucoma treatment is the blood pressure medication losartan. Losartan’s protective activity is due to prevention of remodeling processes that occur in the wall of the eye (the sclera) during glaucoma.

In these studies we have shown that losartan treatment targets specific cells within the sclera called fibroblasts. Fibroblasts exposed to losartan are prevented from becoming “activated” and remodeling the scleral tissue. In addition, we have developed long-lasting, drug releasing microparticles to prevent scleral remodeling in glaucoma.

Carla J. Siegfried, MD
Washington University School of Medicine, St. Louis, MO
Funded by The Alcon Foundation

Project: Pathological Alterations in the Trabecular Meshwork Following Vitrectomy and Lens Extraction: A Model of Oxidative Stress

View and download Dr. Siegfried’s final project research poster.

Summary: Elevation of pressure in the eye is the only risk factor for glaucoma that can be modified. Improved understanding of how the eye’s natural drain is damaged can provide insights to new treatments and prevention of this blinding condition.

We have measured oxygen levels inside the eyes of patients undergoing eye surgery with a small probe and found increased oxygen levels in patients who have had removal of the gel in the back of the eye, a procedure performed for various retinal diseases. Patients who have had this procedure nearly always require cataract surgery and this combination of procedures lead to an increased risk of glaucoma. This excess oxygen may be the source of molecules that cause damage to the cells of the natural drain of the eye. In addition, the level of antioxidants, compounds that protect the cells from this damage, are decreased following this combination of surgeries.

By performing these two procedures (removal of gel and then lens removal), we predicted increased oxygen levels in the front of the eye in the area of the natural drain in a glaucoma model. We were unable to duplicate these findings in this model, but did enhance our techniques utilizing a laser to dissect these specific cells in the drain of the eye to study changes associated with damage and then study the how these cells may have altered programming of their genetic code. In this manner, we can now learn more precisely how these cells are damaged and potentially identify patients who are at risk for damage and new ways to treat glaucoma.

For her research project exploring the role of oxygen and antioxidant levels in the eye, Dr. Siegfried was awarded the 2018 Shaffer Prize for Innovative Glaucoma Research. The Shaffer Prize, presented annually by Glaucoma Research Foundation, recognizes a researcher whose project best exemplifies the pursuit of innovative ideas in the quest to better understand glaucoma.

W. Daniel Stamer, PhD
Duke University Eye Center, Durham, NC
Funded by The Alcon Foundation

Project: Role of Exosomes in Glaucomatous Lamina Cribrosa Remodeling

Summary: In this project we have optimized techniques to isolate and characterize exosomes from lamina cribrosa cells. Exosomes are small vesicles that are released by cells to perform a variety of functions. In the lamina cribrosa, like the trabecular meshwork we hypothesize that exosomes participate in the turnover of extracellular matrix and homeostatic signaling with cell neighbors, particularly in response to elevations in intraocular pressure/pulsations. In the present study we mimicked pressure pulsations by cyclically stretching lamina cribrosa cells and collecting/purifying released exosomes from the cell culture media. We observed that exosomes from glaucomatous lamina cribrosa cells were different than exosomes from normal cells. These differences hold the potential to provide information about abnormal remodeling of the optic nerve head in early stages of glaucoma.

David T. Stark, MD, PhD
Stein Eye Institute, David Geffen School of Medicine at UCLA, Los Angeles, CA
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Endocannabinoids in Retinal Ganglion Cell Regeneration

View and download Dr. Stark’s final project research poster.

Summary: Many optic nerve diseases result in permanent loss of vision. This occurs in part because the intrinsic growth capacity of retinal ganglion cells rapidly declines after birth, and injured central nervous system axons fail to regenerate. The scientific community has learned a great deal about molecular signals that can support regeneration of damaged neural connections, but there is an urgent need to identify as many pro-regenerative signals as possible because it is not clear which of these might actually translate to use in patients.

Generous support from the Glaucoma Research Foundation allowed us to develop a strategy to comprehensively assess an entire class of biomolecules called lipids for differences that occur during optic nerve regeneration. We hope to use this approach to identify candidate molecules that might represent previously unknown pro-growth signals.

Findings from this research study were published in the January 2018 issue of IOVS (Investigative Ophthalmology & Visual Science), “Optic Nerve Regeneration After Crush Remodels the Injury Site: Molecular Insights From Imaging Mass Spectrometry.”

Evan B. Stubbs, Jr., PhD
Edward Hines, Jr. VA Hospital, Hines, IL
Funded by The Alcon Foundation

Project: Mitochondrial-specific Antioxidant XJB-3-151 as a Novel Therapeutic Strategy to Lower Elevated Intraocular Pressure

View and download Dr. Stubbs’ final project research poster [1 of 2].

View and download Dr. Stubbs’ final project research poster [2 of 2].

Summary: Glaucoma is a silent disease that, over time, kills the nerve cells of the retina leading to irreversible blindness. Current treatment options are restricted to non-specific interventions aimed at lowering intraocular pressure (IOP). For many glaucomatous patients, however, pharmacological and surgical management of IOP does not always help. The development of targeted therapeutic strategies directed at the cause of elevated IOP is critical for the advanced management of glaucoma. The cause of elevated IOP most likely involves a molecule called transforming growth factor-β2 (TGF-β2). Funding support from the Glaucoma Research Foundation Shaffer Grant has allowed our lab to advance our understanding of exactly how TGF-β2, a multifunctional cytokine, promotes increases in IOP of patients with POAG. We found that specific cells in the eye, called TM cells, constitutively express and secrete TGF-β2, highlighting the TM as a viable targetable source of TGF-β2. This molecule was further found to elicit harmful and pronounced oxidative stress to the TM. Our findings are consistent with other studies also reporting elevated levels of oxidative stress markers in the eyes of POAG patients, along with altered expression of antioxidant defenses in the TM. Results from this Shaffer Grant study also show that targeting antioxidants such as XJB-5-131 to the TM significantly attenuates expression and release of TGF-β2 from cultured human TM cells. Of equal importance, XJB-5-131 protected human primary TM cells against TGF-β2 mediated changes in expression of specific extracellular matrix proteins. These exciting findings are being put to the challenge to see if targeting antioxidants to the TM in porcine and human eyes will lower IOP. To do this, we have encapsulated small beads, called nanoparticles, with various test agents. We are seeing, for the first time, that these nanoparticles can markedly reduce IOP by reducing endogenous expression of TGF-β2. Collectively, our findings support targeted disruption of constitutive TGF-β2 expression within the eye using antioxidant-encapsulating nanoparticles and raises enthusiasm that this strategy will be a clinically useful and effective new therapy by which to better manage IOP in patients with POAG.

David A. Sullivan, MS, PhD, FARVO
Co-investigator: Louis R. Pasquale, MD
Schepens Eye Research Institute, Massachusetts Eye and Ear, Harvard Medical School, Boston, MA
Funded by the Dr. Henry A. Sutro Family Grant for Research

Project: Estrogen & Glaucoma

View and download Dr. Sullivan’s final project research poster.

Summary: Glaucoma is characterized by a gradual loss of retinal ganglion cells (RGCs), which leads to a loss of vision. The most common form of glaucoma, occurring in 70 to 90% of patients, is primary open angle glaucoma (POAG).

One of the most compelling epidemiological features of POAG is that its incidence shows a striking sex-related difference. Women have a significantly lower incidence of POAG, as compared to men, until the age of 80 years. This sex-related difference has been linked to the extent of lifetime estrogen exposure. Indeed, there is a strong assocation between increased estrogen exposure and a reduced POAG risk. Conversely, studies have shown that a decreased exposure (i.e. early loss of estrogens), confers an increased risk of POAG. We hypothesize that an early estrogen deficiency accelerates the aging of the optic nerve and predisposes this nerve to glaucomatous damage.

To test our hypothesis we determined whether early estrogen deficiency is associated with heightened intraocular pressure, RGC loss and glaucoma in an animal model. Our results demonstrate that estrogen deprivation does promote the development of glaucoma in female mice. To continue these studies, we seek to determine whether estrogen administration will serve as a novel preventive treatment for glaucoma, and in particular, POAG. If so, our research will have significantly advanced our understanding of the role of estrogen in the pathophysiology of glaucoma.

Frank Talke, PhD
University of California, San Diego
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Development of an Optical-based Intraocular Pressure Sensor

View and download Dr. Talke’s final project research poster.

Summary: We have developed an intraocular pressure sensor based on the principle of interferometry. The sensor is comprised of a diaphragm and a glass substrate. By directing monochromatic light towards the active sensing region and applying pressure, one can observe interference fringes as the diaphragm deflects. Using the principle of interferometry, we were able to back calculate pressure using the images captured by a camera. From our studies, we have found the best results using silicon nitride as the diaphragm material. The sensor read-out can be further optimized by coating a thin layer of silicon nitride onto the glass substrate. In order to further demonstrate proof of concept, our sensor was also tested ex-vivo using a rabbit eye model. Thus far, we have achieved 0.8 mmHg resolution. Our results show that the sensor response is repeatable and agree with mathematical models.

2015 Shaffer Grants

Donald L. Budenz, MD, MPH
University of North Carolina, Chapel Hill, NC
Funded by the Dr. Henry A. Sutro Family Grant for Research

Project: Incidence of Glaucoma and Glaucoma Progression in an Urban West African Population

Summary: Glaucoma is the leading cause of irreversible blindness worldwide and disproportionately affects people of African descent because it occurs more frequently, has a younger age of onset, and a more aggressive course than other people groups. These findings are largely based on epidemiology studies performed outside of Africa, specifically the US and the Caribbean. Studies in East Africa and South Africa have found a much lower prevalence of glaucoma than those performed in the US and Caribbean, perhaps because people of African descent residing in the US and Caribbean are descendants of West Africans. Recently, we performed the first properly designed glaucoma prevalence study in West Africa (Ghana) and found a prevalence much more similar to the US and Caribbean populations. We also collected over 1,200 blood sample for genetic analysis, creating the largest genetic database in people of African descent in the world, in an attempt to identify the genes for glaucoma in this people group. The current study is designed to do three things: determine the number of new cases per year (incidence) of glaucoma in this population (no glaucoma incidence studies have been performed in sub-Saharan Africa), determine the rate of progression of glaucoma in the 362 people identified with glaucoma in the original study, and to add to the genetic material in our quest to identify the genes involved in glaucoma in people of African descent.

Paul L. Kaufman, MD
University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research
Co-funded by The Alcon Foundation

Project: Gene Therapy for Glaucoma

Summary: Glaucoma is often associated with elevated intraocular pressure (IOP). At present, the only effective approach to treat glaucoma is to reduce IOP. IOP rises beyond what the eye can tolerate because of increased resistance to fluid leaving the eye in the outflow pathways. There are two main outflow pathways: uveoscleral and trabecular. Prostaglandins, the most commonly prescribed class of glaucoma therapeutics, target the uveoscleral pathway. Compounds are in development to effectively and safely decrease resistance in the trabecular pathway. Self-administration of one or more daily topical medications by patients may affect IOP control due to poor adherence to therapy. Surgical treatments may result in complications and eventual loss of effectiveness, resulting in a return to topical drop therapy. Delivery of therapeutic genes to the eye is a promising strategy to provide long term IOP control, removing the patient from the drug delivery system. In this project, we aim to develop viral vector based therapeutic constructs that target the cytoskeleton of the trabecular meshwork ™, the key structure of the main drainage route. The vectors will be designed to express cytoskeleton-modulating proteins (caldesmon and C3) known to increase open spaces in the TM, thus increasing fluid flow from the eye and reducing IOP. Vectors will be tested in an organ culture system to measure effectiveness. Fluorescent proteins and other markers will be attached to the vectors to enable identification of cell types transfected by the vectors. Successful IOP lowering by the vectors will facilitate development of gene therapy for glaucoma patients.

Richard T. Libby, PhD
University of Rochester Medical School, Rochester, NY
Funded by The Alcon Foundation

Project: Understanding Axonal Degeneration Pathways in Glaucoma

Summary: Loss of vision in glaucoma is caused by the death of a specific type of neuronal cell, the retinal ganglion cell (the neuron that sends information to the brain). Presently there are no treatments aimed at neuroprotection for glaucoma patients. Unfortunately, this means that for many patients, physicians are left with no treatment options to prevent the progression of vision loss. This project aims to determine the molecular signaling pathways responsible for killing retinal ganglion cells in glaucoma. In this application, we concentrate on defining the molecular pathways that control axonal degeneration in retinal ganglion cells after glaucoma-relevant injuries, including ocular hypertension. Specifically, using genetic resources, we will determine whether two molecules that are important for axonal degeneration, prevent retinal ganglion cell death after axonal injury. Given the importance of axonal insult and degeneration in glaucoma, the experiments proposed in this study have the potential to define key therapeutic targets for developing neuroprotective treatments for glaucoma that target early pathological events.

For his research project to explore a novel idea in the field of neurodegeneration, defining the molecular cascade that controls axon degeneration, which is a key early event in glaucoma, Dr. Richard Libby was awarded the 2017 Shaffer Prize from Glaucoma Research Foundation. The Shaffer Prize, presented annually by Glaucoma Research Foundation, recognizes a researcher whose project best exemplifies the pursuit of innovative ideas in the quest to better understand glaucoma.

Paloma Liton, PhD
Duke University Eye Center, Durham, NC
Funded by Dr. James and Elizabeth Wise

Project: Lysosomal Enzymes, Glycosaminoglycans and Outflow Pathway Physiology

Summary: Glaucoma is a group of eye diseases that lead to damage to the optic nerve and can result in irreversible blindness. In the most common form of the disease, damage to the optic nerve is caused by elevated pressure inside the eye, due to a resistance of the aqueous humor to exit the eye. The exact mechanisms leading to that resistance to aqueous humor outflow are not known, but it has been speculated that blockage of the outflow channels might be a contributing factor. Accordingly, patients affected with glaucoma often present a build up of amorphous material in the outflow channels. Genetic studies showed lower amounts of alpha L-Iduronidase (IDUA) in the glaucomatous outflow pathway. IDUA is a lysosomal enzyme that is needed to break down sugars known as glycosaminoglycans (GAGs). These sugars are used to build tissues, but if not properly degraded, they accumulate in the body. IDUA deficiency causes mucopolysaccharidosis, a disease characterized by the accumulation of GAGs inside and outside the cells, gradually leading to tissues and organ dysfunction, with eventual cell death. Patients affected by this disease often develop ocular hypertension and glaucoma. Here, we propose (1) to clarify whether accumulation of GAGs causes elevated ocular pressure by characterizing the functionality of the outflow channels in a murine model of human mucopolysaccharidosis; and (2) test whether exogenous supplementation of IDUA improves outflow pathway tissue function. These studies have the potential to develop novel therapeutic strategies for the treatment of ocular hypertension and glaucoma.

Lyne Racette, PhD
Indiana University, Indianapolis, IN
Funded by the Dr. Miriam Yelsky Memorial Research Grant

Project: Early Detection of Glaucoma Progression using Structural and Functional Data Jointly

Summary: The presence and rate of progression in glaucoma influence clinical decisions, yet the methods currently available to monitor progression are imprecise and do not allow clinicians to make accurate assessments of their patients. We recently developed an innovative model to detect and monitor glaucoma progression. This dynamic structure-function model jointly uses information from the structure and function of the eye to determine whether the disease is progressing. The model is also individualized to each patient to improve its ability to tease out true progression from variability. This is crucial because the large differences that exist between patients can mask the presence of change. The objective of this project is to test our model in the earliest stages of glaucoma. Detecting early changes is crucial to minimize vision loss. Using data from the large Ocular Hypertension Treatment Study, we will assess the specificity and sensitivity of our model in identifying conversion from ocular hypertension to glaucoma. We will also determine whether our model is able to detect this conversion at an earlier point in time. At the conclusion of this study, clinicians will have a powerful method to detect glaucoma progression, leading to improved patient care and preservation of sight.

Matthew A. Smith, PhD
University of Pittsburgh, Pittsburgh, PA
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Measuring the In-vivo Effects on the Optic Nerve Head of Acute Variations in Cerebrospinal Fluid Pressure

Summary: Glaucoma is a leading cause of blindness and visual morbidity worldwide, and yet the pathophysiology of the glaucomatous process is still lacking fundamental understanding even considering recent advancements in imaging technology and genetics. The deleterious effects to the eye of elevated intraocular pressure (IOP) have been known for long and are now considered the main risk factor for glaucoma. A critical barrier for improving glaucoma diagnosis and treatment has been the lack of a complete understanding of the role of IOP in the eye and the causes underlying the range of patient sensitivities to IOP. Eyes that demonstrate similar clinical features may react differently to changes in IOP. The reason for these differences is mostly unknown. Our global hypothesis is that both IOP and the pressure inside the brain (cerebrospinal fluid pressure – CSFP) are significant contributors to the biomechanical environment within the optic nerve head. Hence, the sensitivity to IOP of a particular subject can be better predicted by considering CSFP. Our research aims to measure and manipulate IOP and CSFP in vivo in an animal model in order to uncover the factors that drive different sensitivity to IOP in different eyes. Our work will establish the fundamental principles by which the pressures inside the eye and the brain interact, and provide an avenue for understanding and eventually treating glaucoma by taking all properties of each individual eye into account.

Shandiz Tehrani, MD, PhD
Oregon Health & Science University, Portland, OR
Funded by The Alcon Foundation

Project: Local Drug Delivery to the Optic Nerve Head as a Novel Treatment in Experimental Glaucoma

Summary: Glaucomatous damage to axons occurs at the optic nerve head (ONH). Support cells within the ONH, called astrocytes, provide multiple functions to protect axons. However, early ONH astrocyte activation has been identified as a potential source of axonal injury in glaucoma. Strategies acting to maintain normal astrocyte function may lead to preservation of ONH axons and therefore reduce glaucomatous damage. The development of targeted drug-delivery strategies to sustain ONH astrocyte structure and function is an important area of research. ONH astrocytes have cellular extensions, which ensheath axons. We have shown that these cellular extensions are rich in a cytoskeletal protein called actin and re-orient prior to axonal injury in a rat model of glaucoma. The objective of this proposal is to locally deliver small molecules to the ONH in a rat model of glaucoma, with a specific aim of determining if local ONH actin modulation in vivo will alter ONH axonal survival. Our research is based upon the overall hypothesis that normal ONH actin-rich astrocyte extensions are necessary for axon survival, and disruption of astrocyte actin assembly will be detrimental to axons. Through our work, we will identify a novel ONH drug delivery method which can be used to assay other molecular pathways that may be involved in axon injury, and to test local axon-protective effects of small molecules in experimental glaucoma.

Gülgün Tezel, MD
Columbia University, New York, NY
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Molecular Biomarkers of Glaucoma

Summary: Glaucoma is a leading cause of blindness affecting millions of Americans. However, current treatment strategies are not sufficient to prevent disease progression and no specific blood test is available for early diagnosis and better follow-up of this blinding disease. To accomplish better management of glaucoma, our experimental research aims to characterize disease-causing molecular alterations and identify molecules that can be used for clinical testing. Our recent studies have indicated four specific molecules (apoptosis-inducing factor, CREB-binding protein, ephrin type-A receptor, and huntingtin protein) that can be measured in blood samples and exhibit increased levels in patients with glaucoma. The proposed project aims to determine the value of these molecules for clinical testing in glaucoma. We will therefore analyze the presence and abundance of these molecules (called “oecandidate biomarkers”) in blood and aqueous humor (intraocular fluid that fills the space between the cornea and the iris) samples collected from larger groups of patients with or without glaucoma and age-matched controls, and determine their predictive value for the initiation and progression of glaucoma. We expect that this new project will provide important information about specific molecular markers (called “biomarkers”) to diagnose glaucoma early, predict its prognosis, and monitor disease progression and treatment responses in patients with glaucoma. Prediction and early diagnosis of glaucoma will allow early treatment to halt disease progression, and monitoring the disease progression and treatment responses will facilitate the efforts ongoing to develop new and improved treatments for glaucoma.

In September 2015, IOVS (vol. 56 no. 10) published results from this research project in “Proteomics Analysis of Molecular Risk Factors in the Ocular Hypertensive Human Retina.” The published paper concluded that” “molecular alterations detected in the ocular hypertensive human retina as opposed to previously detected alterations in human donor retinas with clinically manifest glaucoma suggest that proteome alterations determine the individual threshold to tolerate the ocular hypertension-induced tissue stress or convert to glaucomatous neurodegeneration when intrinsic adaptive/protective responses are overwhelmed.”

2014 Shaffer Grants

Jeff M. Gidday, PhD
Washington University School of Medicine, St. Louis, Missouri
Funded by the Dr. Miriam Yelsky Memorial Research Grant

Project: Delayed Post-Conditioning for Glaucoma Neuroprotection

Summary: Protection of retinal neurons that die in glaucoma is a fundamental therapeutic strategy, but one that remains elusive. Although basic research documents that these cells die by a multi-factorial process, the vast majority of therapies tested to date, or in development, are likely to fail because they target only a single injury pathway. A novel way of approaching protection-based therapeutics for glaucoma should derive from evidence accumulating over two decades in stroke and cardiac arrest: That simultaneously activating a variety of self-defense responses in cells with stressful “conditioning” stimuli induces the expression of a host of genes that promote cell survival. We plan to test two such “epigenetics”-based therapeutic strategies in a model of glaucoma. If successful, our studies will provide a viable therapeutic strategy for saving vision in patients with glaucoma.

Vikas Gulati, MD
Truhlsen Eye Institute, University of Nebraska Medical Center, Omaha, Nebraska
Funded by The Alcon Foundation

Project: Effect of Vascular Endothelial Growth Factor Blockers on Aqueous Humor Dynamics

Summary: Use of eye injections of drugs aimed at slowing the growth of abnormal blood vessels in the eye is becoming more common for the treatment of individuals with many eye problems including macular degeneration, diabetic retinopathy and retinal artery or vein blockage. Unfortunately, injections of these drugs can cause high eye pressure that may lead to glaucoma in some patients or make the control of high eye pressure more difficult in others. Considering the large number of people over the age of 40 who have glaucoma, many may be at particular risk of permanent vision loss from loss of pressure control when treated with these injections. Eye pressure by itself is determined by a delicate balance of fluid inflow and outflow from inside the eye. The effect of the injected drugs on any of these parameters has never been formally evaluated. The purpose of this study is to determine the effects of these drugs on the inflow and outflow of fluid from the eye and its consequences on eye pressure. Knowledge of the actual physiological effect of these drugs can have many benefits for glaucoma patients. For one it will help the eye doctor choose the optimal therapy to prevent the eye pressure elevation or to treat the high eye pressure if it does occur. Further research also will help determine which if any of these drugs is safer for glaucoma patients who also have abnormal blood vessel growth.

David Krizaj, PhD
Moran Eye Institute, University of Utah, Salt lake City, Utah
Funded by Dr. James and Elizabeth Wise

Project: RGC Mechanotransduction as a Target in Glaucoma

Summary: The major diagnostic criterion for glaucoma is elevated pressure in the eye which identifies a proportion of patients that will develop this blinding disease. However, pressure-reducing drugs often help patients with elevated and normal intraocular pressure (IOP). The mechanism that is responsible for transducing the effects of eye pressure into degeneration of retinal ganglion cells (RGCs) is not known. Our proposal addresses this problem. We found that RGCs are the sole retinal neuron that express TRPV4, a pressure-sensitive channels that is permeable to calcium ions. This is intriguing because calcium is known to induce remodeling of cells and RGC degeneration in glaucoma. We will use molecular, histological, calcium imaging and electrophysiological methods to characterize the sensitivity of these channels to membrane stretch by mimicking the effect of IOP on RGC perikarya and axons. Second, we will characterize the effect of stretch on cells isolated from mouse glaucoma models. Third, we will study the effect of small molecule antagonist drugs to prevent stretch-mediated RGC degeneration and death. The study is strongly supported by preliminary evidence obtained in vitro and in vivo. Thus, the proposed work directly tackles the mechanosensitive disease mechanism in the posterior eye but also aims at developing new neuroprotective strategies.

Yutao Liu, MD, PhD
Medical College of Georgia, Georgia Regents University, Augusta, Georgia
Funded by The Alcon Foundation

Project: Exosomal RNAs and Aqueous Humor Dynamics

Summary: The purpose of this study is to investigate the role of exosomes, small cell-manufactured vesicles secreted into bodily fluids including blood and the fluid in the eye called aqueous humor. Exosomes contain RNA, which is used to regulate cell function. Secreted RNAs have already been shown to be involved in cell-cell communication between different tissues/organs and to serve as biomarkers for human disorders, such as cancer. We will examine the characteristics of the RNA contained in exosomes in the aqueous humor with special emphasis on its function in exfoliation glaucoma, including the most common form of secondary open-angle glaucoma. This study may provide a potential biomarker and therapeutic targets for glaucoma.

Dr. Liu’s research results from this study were published in the January 18, 2015 edition of the peer-reviewed journal Experimental Eye Research, and the February 1, 2018 issue of the journal Human Molecular Genetics.

Stuart J. McKinnon, MD, PhD
Duke University Medical Center, Durham, North Carolina
Funded by Dr. James and Elizabeth Wise

Project: Neuroinflammation: The Role of Lymphocytes in Glaucoma

Summary: In glaucoma, permanent vision loss and blindness occur when retinal ganglion cells (RGCs) that make up the optic nerve are lost. Increasing evidence points to a central role of the immune system in the death of RGCs in glaucoma. A recent finding in our laboratory led to the novel hypothesis that immune system events involving lymphocytes are necessary for RGC cell death and optic nerve axon loss in glaucoma. This project will determine whether specific populations of lymphocytes are required for RGC death in glaucoma. Based on findings from this study, therapies might eventually be designed to modulate the immune system in order to prevent vision loss and blindness in glaucoma patients.

For his research project to determine whether therapies can be designed to modulate the immune system to prevent vision loss and blindness in glaucoma patients, Stuart J. McKinnon, MD, PhD was awarded the 2016 Shaffer Prize for Innovative Glaucoma Research. The Shaffer Prize, presented annually by Glaucoma Research Foundation, recognizes a researcher whose project best exemplifies the pursuit of innovative ideas in the quest to better understand glaucoma.

Robert W. Nickells, PhD
University of Wisconsin, Madison, Wisconsin
Funded by the Dr. Henry A. Sutro Family Grant for Research

Project: Purinergic Signaling of Neuroinflammatory Glial Responses in a Model of Optic Nerve Damage

Summary: The central nervous system (CNS) is made up of neuronal cells and support cells, called glia. Whenever there is damage to the CNS, such as in the retina during glaucoma, nerve cells die and glial cells change their behavior in a process called activation. Although glial activation may have short-term benefits, there is consensus that in the long-term, these cells may contribute to the pathology of damaged neurons in glaucoma. Currently, we know that glia become activated in glaucoma, but we do not know by what process. More and more evidence from other parts of the brain suggest that a molecule called ATP is part of the signaling mechanism where damaged neurons tell the glia that they are in distress. We have preliminary evidence that this is also true in the retina after damage to the optic nerve. This proposal is aimed at determining if dying neurons in the retina (ganglion cells) signal to the retinal glia by releasing ATP through specialized channels made up of a protein called Pannexin1. An increased understanding of how the neurons and glia communicate in glaucoma should lead to new developments on how to “tame” the glia into a non-pathologic role, thus increasing the potential for greater preservation of sight.

Colm O’Brien, MD, FRCS
Mater Misericordiae University Hospital, Dublin, Ireland
Funded by The Alcon Foundation

Project: Caveolins, Calcium Signalling and Fibrosis of Lamina Cribrosa Cells in Glaucoma

Summary: Glaucoma is the second most common cause of vision loss and blindness in the world. Patients with glaucoma present at the clinic with loss of peripheral vision and eye pressure often above normal levels causing compression and damage in a part of the optic nerve called the lamina cribrosa (situated at the back of the eye). We hope to determine if proteins present in lamina cribrosa of glaucoma patients differ to those found in people who do not have the disease; as these proteins may contribute to disease progression. We are interested in three classes of proteins; one which is responsible for the hardening of the cells and their surrounding environment (these are known as fibrotic proteins) in glaucoma and the other which regulates the level of calcium entering and exiting cells. We hypothesise that a third protein group, the caveolin scaffolding proteins, may provide a link between fibrosis and calcium levels and may be responsible for their disregulation in glaucoma. It is our hope that the long-term outcome of this project will be a strategy for relieving the disease burden for sufferers of glaucoma.

Joshua D. Stein, MD, MS
W.K. Kellogg Eye Center, University of Michigan, Ann Arbor, Michigan
Funded by the Glaucoma Research Foundation Board of Directors

Project: A Dynamic, Personalized Glaucoma Monitoring Decision Support Tool

Summary: Our goal is to develop a powerful new type of glaucoma decision support tool to help eye doctors quickly and effectively identify which glaucoma patients are at high risk of getting worse and prevent them from losing more vision. Key features of this innovative technology are that it (1) learns more and more about the stability of the patient’s glaucoma with every measurement of eye pressure or visual field, (2) can be personalized to identify the optimal frequency of testing for each individual patient, and (3) would suggest an eye pressure level specific for that particular patient that the care provider would use in the process of recommending treatment. Preliminary results from an early version of this technology demonstrate that it is capable of identifying patients whose glaucoma is getting worse 57% sooner than existing approaches and it is able to significantly reduce the number of eye pressure and visual field measurements required to check for glaucoma worsening. This grant will enable us to advance and expand our technology so that it (1) determines for each patient the ideal level of eye pressure, and (2) guides eye doctors about how frequently each patient should undergo various glaucoma tests like checking eye pressure or undergoing visual field testing. With Glaucoma Research Foundation support, we plan to create this new methodology for a glaucoma decision support tool and further test it so that it will soon be ready to help eye doctors prevent their patients with glaucoma from getting worse.

2013 Shaffer Grants

Anneke I. den Hollander, PhD
Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
Funded by the Dr. Miriam Yelsky Memorial Research Grant

Project: Dissecting the Genetic Causes of Congenital and Juvenile Glaucoma

Summary: Glaucoma is a leading cause of irreversible blindness affecting 70 million people worldwide. There are various types of glaucoma, and two of them can affect children and young adults: primary congenital glaucoma (PCG) and juvenile open-angle glaucoma (JOAG). PCG and JOAG are hereditary diseases that can be inherited in families. The genetic causes of PCG and JOAG partially overlap with adult-onset primary open-angle glaucoma (POAG), the most common form of glaucoma. We believe that a significant proportion of the genetic causes of POAG can be explained by genetic variants in genes that underlie PCG and JOAG. In this study we aim to identify new genetic causes of PCG and JOAG using the newest genetic technologies (exome sequencing) in families, and to evaluate the role of such new genes in POAG patients. The results of this project will improve our understanding of glaucoma, which will enable the design of new therapies.

John H. Fingert, MD, PhD
University of Iowa, Department of Ophthalmology and Visual Sciences, Iowa City, Iowa
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Molecular Genetic Study of Normal Tension Glaucoma using Transgenic Mice

Summary: There is a critical need to clarify the mechanisms of glaucoma at the molecular level to help provide physicians with tools for early detection and treatment. Recently, we showed that duplication of a gene (TBK1) causes some cases of a form of glaucoma that occurs at low eye pressure. Some patients have glaucoma because they carry an extra copy of TBK1 in their genome. We plan to extend this discovery by developing a model of TBK1 glaucoma that will facilitate studies of the basic mechanisms by which defects in genes cause the disease. The project may also facilitate development and testing of new sight-saving drug therapies for glaucoma.

In February 2015, Ophthalmology Times reported that Dr. Fingert’s continuing research provides strong evidence that mutation of the TBK1 gene can lead to glaucoma and may provide insights into disease mechanisms and future treatments. “Hopefully this will open up a new field for low-pressure glaucoma research and treatment,” Dr. Fingert said. He presented his research results at the 2014 meeting of American Academy of Ophthalmology.

M. Elizabeth Fini, PhD
University of Southern California, Institute for Genetic Medicine, Los Angeles, California
Funded by the Merck Department of Continuing Education

Project: Novel Mucins and Aqueous Outflow

Summary: All forms of glaucoma have in common optic nerve degeneration characterized by typical visual field defects, and are usually associated with elevated intraocular pressure, also known as ocular hypertension (OH). In most instances, OH results from impaired drainage of aqueous humor through the trabecular meshwork ™. Treatment with steroid drugs in the eye can cause OH insusceptible individuals. In preliminary studies, two newly discovered genes encoding sugary molecules called mucins were associated with steroid-induced OH. It is hypothesized that the novel mucins are part of a sugar-rich TM coating known as the glycocalyx, and that their altered production in response to steroids could lead to OH. The purpose of this project is to provide additional supporting data. Behavior of the two novel mucins will be examined in cultured TM cells and in the TM of intact eyes using recombinant DNA, biochemical, and imaging techniques in order to provide clues to function. Glycocalyces are found in all organs and play important roles in health and disease. Recent studies suggest that the glycocalyx in the outflow pathways of the eye may be much more extensive than previously imagined. The idea that mucins might be present in this lining layer and play a role in OH has not been previously considered. If confirmed, the findings will open a new line of research that could ultimately lead to significant innovation, as drugs that control amounts of the novel mucins could be a new treatment paradigm for glaucoma.

Andras M. Komaromy, DrMedVet, PhD
Michigan State University, East Lansing, Michigan
Funded by The Alcon Foundation

Project: Gene Therapy in a Spontaneous Canine Model of Primary Open-Angle Glaucoma

Summary: Primary open-angle glaucoma (POAG) is a leading cause of incurable blindness. Increased pressure inside the eye due to slowed fluid drainage contributes to the disease process in a majority of patients with POAG. Because some families seem to be affected more than others, inherited risk factors are suspected to play an important role in the development of glaucoma. Indeed, several genetic defects have been identified that likely contribute to the pressure increase inside the eye. In this project we intend to show that we can insert healthy copies of a damaged gene into the fluid drainage channels inside the eye and normalize eye pressure. Our project will hopefully provide proof of principle that gene therapy could one day provide lasting control of normal eye pressure in patients with known genetic defects.

For his research on the potential of gene therapy to provide lasting control of intraocular pressure in glaucoma patients with known genetic defects, András Komáromy, DVM, PhD was awarded the 2015 Shaffer Prize for Innovative Glaucoma Research. The Shaffer Prize, presented annually by the GRF Scientific Advisory Committee, recognizes a researcher whose project best exemplifies the pursuit of innovative ideas in the quest to better understand glaucoma. Dr. Komáromy studies the molecular causes of inherited eye diseases in dogs and is working to develop gene therapies to stop vision loss. By identifying and treating gene mutations in dogs, his research moves us closer to gene therapy that could one day be used to manage and prevent glaucoma in humans.

Colleen M. McDowell, PhD
University of North Texas Health Science Center, Fort Worth, Texas
Funded by The Alcon Foundation

Project: Retina Ganglion Cell Subtype Specific Cell Death in a Mouse Model of Human Primary Open-Angle Glaucoma

Summary: The goal of this project is to better understand the mechanisms involved in glaucomatous injury to the eye. We will study specific subtypes of cells in the eye that are more or less susceptible to glaucoma damage. Damage to the visual sensing structures in the eye and brain will be evaluated over time in order to determine onset and extent of damage. This project will help identify pathways that may serve as new targets for the development of effective glaucoma treatments. These experiments also may lead to the discovery of more sensitive ways to diagnose glaucoma and follow glaucoma progression.

Yvonne Ou, MD
University of California San Francisco, Department of Ophthalmology, San Francisco, California
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Investigating Axonal Death Pathways in Glaucoma

Summary: A major deficit in glaucoma management is that a diagnosis is made or treatment is initiated after there is already evidence of optic nerve cell death or visual field loss. Our goal is to investigate the parts of the optic nerve cell, specifically axons and synapses, which may be vulnerable early in the course of the disease. Axons are the long projections of neurons that conduct electrical impulses, and information is transmitted from one neuron to a second neuron across the synapses located at the ends of neurons. Investigation of the effects of elevated eye pressure on the optic nerve cell axons and synapses is critical to our long-term goals of improving diagnosis and treatment for glaucoma patients. We will use a glaucoma model to study whether the gene Sarm1 plays a role in glaucoma-induced axon death and synapse loss in the retina and brain. If Sarm1 plays a role in axon or synapse loss in our model, it would be an attractive drug target for treating glaucoma. This project seeks to uncover a new approach to glaucoma diagnosis and treatment before the optic nerve is irreversibly damaged.

David Sretavan, MD, PhD
University of California San Francisco, San Francisco, California
Funded by the Frank Stein and Paul S. May Grants for Innovative Glaucoma Research

Project: Pathophysiological Progression in Single RGC Axons Following Microscale Compressive Injury

Summary: The debilitating loss of vision associated with advanced forms of glaucoma result directly from the degeneration of Retinal Ganglion Cells (RGC) in the retina. The pattern of RGC loss in patients as well as information obtained from laboratory research all point to the fact that an important site of pathology occurs at the optic nerve head, a region where the axonal cell processes of RGCs exit the eye on their way to the visual centers of the brain. Compressive injury associated with the elevated eye pressures in glaucoma is thought to exert a deleterious effect directly on RGC axons at this site, eventually compromising the normal biological processes required for overall RGC health, and ultimately leading to RGC death. Despite this generally well-accepted idea for how high eye pressure may affect RGC axons, scientists still do not understand the injury mechanisms involved in sufficient detail to begin identifying potential therapeutic targets. A major hurdle in elucidating the progression of axon pathology is the lack of research instrumentation to systematically map the effects of compressive injury on individual nerve cell elements. Our project will utilize two novel microscale technologies originating from our laboratory, namely highly precise molecular micropatterning and miniaturized axon nano-compressors to address this problem. Results from this study may allow us to better understand the injury threshold leading to irreversible RGC degeneration. This in turn can provide insight into the key cellular pathways that are potentially amenable for therapeutic intervention.

Lin Wang, MD, PhD
Devers Eye Institute/Legacy Research Institute, Portland, Oregon
Funded by The Alcon Foundation

Project: Noninvasive Assessment of Dynamic Autoregulation in Optic Nerve Head

Summary: Glaucoma is one of the leading causes of blindness worldwide characterized by irreversible damage in the optic nerve head (ONH). Yet, the causes to the ONH damage remain unclear. One possible theory underlying the mechanism is that the blood supply to the ONH in glaucoma patients becomes insufficient due to impaired “autoregulation” capacity, an intrinsic function of a tissue to maintainconstant blood supply. However, this theory has never been conclusively proven due partially to a lack of effective methods to quantify the performance of autoregulation in the ONH. In this study, a new method to assess the performance of autoregulation in ONH is proposed. It takes advantage of spontaneous fluctuation in blood pressure (BP) and artificially induced BP change. The ultimate goal is to use the methods and analytical techniques to examine the ONH autoregulation capacity and to define the autoregulation abnormalities in glaucoma.