Aim 1. Establish interventions for early disease in the transgenic rhodopsin P347L (TgP347L) rabbit, a model of human retinitis pigmentosa (RP). TgP347L 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. Significance. This is a test of early intervention for human RP in 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. Significance. 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 implications 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 and- 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 remodeling RP in a large-eye animal model.

Public Health Relevance

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 effectiveness of gene therapies.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY015128-08
Application #
8294249
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
2012-04-01
Budget End
2013-03-31
Support Year
8
Fiscal Year
2012
Total Cost
$627,106
Indirect Cost
$187,466
Name
University of Utah
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
009095365
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
Marc, Robert E; Anderson, James R; Jones, Bryan W et al. (2014) The AII amacrine cell connectome: a dense network hub. Front Neural Circuits 8:104
Marc, Robert; Pfeiffer, Rebecca; Jones, Bryan (2014) Retinal prosthetics, optogenetics, and chemical photoswitches. ACS Chem Neurosci 5:895-901
Sher, Alexander; Jones, Bryan W; Huie, Philip et al. (2013) Restoration of retinal structure and function after selective photocoagulation. J Neurosci 33:6800-8
Lauritzen, J Scott; Anderson, James R; Jones, Bryan W et al. (2013) ON cone bipolar cell axonal synapses in the OFF inner plexiform layer of the rabbit retina. J Comp Neurol 521:977-1000
Enriquez-Algeciras, Mabel; Ding, Di; Mastronardi, Fabrizio G et al. (2013) Deimination restores inner retinal visual function in murine demyelinating disease. J Clin Invest 123:646-56
Marc, Robert E; Jones, Bryan W; Watt, Carl B et al. (2013) Retinal connectomics: towards complete, accurate networks. Prog Retin Eye Res 37:141-62
Lin, Yanhua; Jones, Bryan W; Liu, Aihua et al. (2012) Rapid glutamate receptor 2 trafficking during retinal degeneration. Mol Neurodegener 7:7
Ross, Jason W; Fernandez de Castro, Juan P; Zhao, Jianguo et al. (2012) Generation of an inbred miniature pig model of retinitis pigmentosa. Invest Ophthalmol Vis Sci 53:501-7
Lin, Yanhua; Jones, Bryan W; Liu, Aihua et al. (2012) Retinoid receptors trigger neuritogenesis in retinal degenerations. FASEB J 26:81-92
Marc, Robert E; Jones, Bryan W; Lauritzen, J Scott et al. (2012) Building retinal connectomes. Curr Opin Neurobiol 22:568-74

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