Aim 1. Establish interventions for early disease in the transgenic rhodopsin P347L (TgP347L) rabbit, a model of human retinitis pigmentosa (RP). TgP347L (1-30 month old) rabbits (1-30 month old) will receive subretinal injections of shRNA-adeno-associated virus (AAV) to suppress mutant opsin production. Analyses include eye imaging, electroretinograms, visual evoked po- tentials, behavior, immunocytochemistry, Western blots, automated transmission electron micro- scope (ATEM) imaging, and electrophysiology by whole cell and multielectrode arrays. Signifi- cance. This is a test of early intervention for human RP in a large-eye animal model.
Aim 2. Establish optogenetic interventions for late disease in RP. TgP347L rabbits >12mos of age will receive (1) subretinal injections of Channelrhodopsin2 (ChR2)-AAV targeting bipolar cells (BCs) or AII amacrine cell (ACs). Analyses will follow those established in Aim 1. Signifi- cance. This is a test of late intervention for human RP in a large-eye animal model.
Aim 3. Build pathoconnectomes for early and late stage TgP347L rabbit BCs and AII cells. BCs are known to undergo severe rewiring and reprogramming in RP, but the nature of retargeting is completely unknown. The extent of AII cell remodeling is similarly unknown. Methods. ATEM imaging will be used to characterize remodeling networks. Significance. This is the first attempt to quantitatively define rewiring in any neural system in retina or brain and has profound implica- tions for all neurodegenerative disorders.
Aim 4. Evaluate retinoic acid and retinoid receptor-based interventions to attenuate corruptive remodeling and reprogramming. TgP347L rabbits will be treated with retinoid X receptor an- tagonists to block pathologic neuritogenesis. Analysis includes ATEM imaging and molecular profiling as in Aim 1. Significance. This is a test of a potential clinical intervention for remodel- ing RP in a large-eye animal model.

Public Health Relevance

EY015128 New RNA inhibition and optogenetics interventions offer potential cures for retinitis pigmentosa (RP), even in the presence neuronal remodeling that impedes other therapies. Discovery of the mechanisms remodeling are facilitating anti-remodeling drug testing that may enhance the ef- fectiveness of gene therapies.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY015128-09
Application #
8451298
Study Section
Special Emphasis Panel (ZRG1-CB-G (02))
Program Officer
Shen, Grace L
Project Start
2003-12-01
Project End
2016-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
9
Fiscal Year
2013
Total Cost
$536,248
Indirect Cost
$180,340
Name
University of Utah
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
009095365
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
Jones, B W; Pfeiffer, R L; Ferrell, W D et al. (2016) Retinal remodeling in human retinitis pigmentosa. Exp Eye Res 150:149-65
Lauritzen, J Scott; Sigulinsky, Crystal L; Anderson, James R et al. (2016) Rod-cone crossover connectome of mammalian bipolar cells. J Comp Neurol :
Woodell, Alex; Jones, Bryan W; Williamson, Tucker et al. (2016) A Targeted Inhibitor of the Alternative Complement Pathway Accelerates Recovery From Smoke-Induced Ocular Injury. Invest Ophthalmol Vis Sci 57:1728-37
Jones, Bryan W; Pfeiffer, Rebecca L; Ferrell, William D et al. (2016) Retinal Remodeling and Metabolic Alterations in Human AMD. Front Cell Neurosci 10:103
Merriman, Dana K; Sajdak, Benjamin S; Li, Wei et al. (2016) Seasonal and post-trauma remodeling in cone-dominant ground squirrel retina. Exp Eye Res 150:90-105
Loizos, Kyle; RamRakhyani, Anil Kumar; Anderson, James et al. (2016) On the computation of a retina resistivity profile for applications in multi-scale modeling of electrical stimulation and absorption. Phys Med Biol 61:4491-505
Pfeiffer, Rebecca L; Marc, Robert E; Kondo, Mineo et al. (2016) Müller cell metabolic chaos during retinal degeneration. Exp Eye Res 150:62-70
Bernstein, Paul S; Li, Binxing; Vachali, Preejith P et al. (2016) Lutein, zeaxanthin, and meso-zeaxanthin: The basic and clinical science underlying carotenoid-based nutritional interventions against ocular disease. Prog Retin Eye Res 50:34-66
Molnár, Tünde; Yarishkin, Oleg; Iuso, Anthony et al. (2016) Store-Operated Calcium Entry in Müller Glia Is Controlled by Synergistic Activation of TRPC and Orai Channels. J Neurosci 36:3184-98
Jiang, Li; Wei, Yuxiao; Ronquillo, Cecinio C et al. (2015) Heterotrimeric kinesin-2 (KIF3) mediates transition zone and axoneme formation of mouse photoreceptors. J Biol Chem 290:12765-78

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