Author:
Kandoi Sangeetha,Martinez Cassandra,Chen Kevin Xu,Reddy L Vinod K.,Mehine Miika,Mansfield Brian C.,Duncan Jacque L.,Lamba Deepak A.
Abstract
AbstractRetinitis 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 ∼25% 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) inRHOas a novel cause of adRP. We (1) show advanced retinal degeneration in a male patient (60-70 year old) harboring four transcriptionally active intact copies of rhodopsin, (2) recapitulated the clinical phenotypes using retinal organoids, and (3) assessed the utilization of a small molecule, Photoregulin3 (PR3), as a clinically viable strategy to target and modify disease progression in RP patients associated withRHO-CNV. Patient retinal organoids showed photoreceptors dysgenesis, with rod photoreceptors displaying stunted outer segments with occasional elongated cilia-like projections (microscopy); increasedRHOmRNA expression (qRT-PCR and bulk RNA-sequencing); and elevated levels and mislocalization of rhodopsin protein (RHO) within the cell body of rod photoreceptors (western blotting and immunohistochemistry) over the extended (300-days) culture time period when compared against control organoids. Lastly, we utilized PR3 to targetNR2E3, an upstream regulator ofRHO, to alterRHOexpression and observed a partial rescue of RHO protein localization from the cell body to the inner/outer segments of rod photoreceptors in patient organoids. These results provide a proof-of-principle for personalized medicine and suggest thatRHOexpression requires precise control. Taken together, this study supports the clinical data indicating that adRP due toRHO-CNV develops due protein overexpression overloading the photoreceptor post-translational modification machinery.
Publisher
Cold Spring Harbor Laboratory
Cited by
1 articles.
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