Disease modeling and pharmacological rescue of autosomal dominant Retinitis Pigmentosa associated with RHO copy number variation.

Abstract

Retinitis pigmentosa (RP), a heterogenous group of inherited retinal disorder causes slow progressive vision loss with no effective treatments available. Mutations in the rhodopsin gene (RHO), account for ~40% cases of autosomal dominant RP (adRP). In this study, we describe the disease characteristics of the first ever reported mono-allelic copy number variation (CNV) in RHO as a novel cause of adRP. We (1) show advanced retinal degeneration in a male patient (~age in years, 60s) harboring four transcriptionally active intact copies of rhodopsin, (2) recapitulate the clinical phenotypes using retinal organoids, and (3) assess the utilization of a small molecule, Photoregulin3 (PR3), as a clinically viable strategy to target and modify disease progression in RP patients associated with RHO-CNV. Patient retinal organoids showed outer segment developmental defects (microscopy), increased RHO mRNA levels (qRT-PCR and bulk RNA-sequencing), along with elevated expression and mislocalization of rhodopsin protein (RHO) within the cell body of rod photoreceptors (western blotting and immunohistochemistry). Lastly, by targeting the upstream regulator of RHO, NR2E3, we effectively altered RHO expression, leading to the partial rescue of RHO protein localization from the soma to the inner/outer segments of rod photoreceptors and providing a proof-of-principle for personalized medicine. Taken together, this study supports the clinical data indicating that adRP due to rhodopsin CNV develops due to a dominant negative gain of function.

Competing Interest Statement

Commercial relationships disclosures: Sangeetha Kandoi - None Cassandra Martinez - None Kevin Xu Chen - None Brian Mansfield - None Jacque L. Duncan - None Deepak A. Lamba - None

Funding Statement

The research presented here is supported by NEI R01 EY032197 (DAL), U24 EY029891 (DAL and JLD), UCSF Vision Core NIH/NEI P30 EY002162, Foundation Fighting Blindness (DAL), All May See Foundation post-doctoral fellowship (SK), and an unrestricted grant from Research to Prevent Blindness, New York, NY

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Institutional Review Board (IRB) of the University of California, San Francisco gave ethical approval for this work.

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Data Availability

All data produced in the present study will be available upon reasonable request to the authors.

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