PROJECT SUMMARY Glaucoma blinds through degeneration of retinal ganglion cells (RGCs) and their axons in the optic projection through sensitivity to intraocular pressure (IOP). Many patients continue to lose vision despite efforts to manage IOP. Thus, an unmet clinical need is a treatment that addresses RGC degeneration directly. Our long-term goal is to identify new therapeutic targets based on neuronal repair, protection, and restoration. In the previous grant cycle, we leveraged transgenic mouse strains to discern interplay between RGC dendritic pruning, axon degeneration, and astrocyte glia. We discovered two novel forms of adaptive remodeling that boost and preserve RGC signaling and slow progression. With unilateral IOP elevation, metabolic redistribution transfers metabolites from the unstressed optic nerve to the retina and nerve challenged by IOP elevation through astrocyte networks. Conditional knock-out of the gap junction protein connexin 43 (Cx43) uncouples this network and prevents redistribution. Finally, for individual RGCs exposed to elevated IOP, enhanced excitability amplifies the light response, even as dendritic complexity diminishes, through reorganization of voltage-gated sodium channels (NaV) in the unmyelinated axon segment. Both phenomena occur early and are transient, as are their protective effects. Our objective in this competitive renewal is to build upon these important results to discern how enhanced excitability and metabolic redistribution mechanistically relate to axonal and dendritic degeneration and whether they can be enhanced to extend RGC survival. As a corollary, we will test whether the transient nature of both forms of adaptation arises from metabolic and oxidative stress to the astrocyte network and if boosting resources exogenously reduces this stress and extends visual function. This hypothesis is supported by new preliminary data showing a dietary metabolite (pyruvate) increases astrocyte glycogen in the optic nerve and enhances nerve excitation in response to elevated IOP, suggesting that the two forms of adaptive remodeling may be linked. In our inducible glaucoma models, we will utilize a cross-disciplinary approach that combines electrophysiological, cellular and in vivo imaging, and transgenic tools. Aim 1 will determine the dependence of adaptive remodeling on axonopathy and dendritic pruning. Aim 2 will characterize the interdependence between metabolic redistribution and enhanced excitability and whether metabolic redistribution through astrocyte networks maps retinotopically to spatial sectors of intact RGC axon and dendritic function. Finally, Aim 3 will test whether boosting metabolic resources reduces astrocyte stress, extends adaptive remodeling, and slows progression in mouse and non- human primate models of glaucoma. Building from results in the prior grant period, our innovative strategy will elucidate how two novel, intrinsically compensatory adaptive processes utilize metabolic resources to promote RGC survival in glaucoma. By translating results to our non-human primate model, we will test the therapeutic value of targeting enhanced excitability and metabolic redistribution as clinical interventions.

Public Health Relevance Glaucoma is the leading age-related cause of irreversible blindness worldwide and will afflict an estimated 112 million people by 2040. The disease causes retinal and optic nerve degeneration through sensitivity to ocular pressure, but lowering pressure does not always stop progression. The work proposed here will test how adaptive mechanisms in the retina and optic nerve could slow degeneration and preserve vision in glaucoma by distributing energy resources where needed to enhance signaling from the retina to the brain.