Allele-selective reduction of P23H-mutant rhodopsin with stereopure oligonucleotides rescues phenotype associated with retinitis pigmentosa in preclinical models

Abstract

Purpose : We tested whether a stereopure antisense oligonucleotide that selectively targets the P23H mutation in Rhodopsin (RHO) but that spares expression of healthy RHO transcripts can address phenotypes associated with the autosomal dominant form of retinitis pigmentosa (adRP) that is caused by this mutation.

Methods : We applied PRISM^TM, Wave’s proprietary discovery and drug development platform, to generate stereopure antisense oligonucleotides that selectively target the RHO P23H mutation. We evaluated allele-selective activity of the oligonucleotides against RHO alleles with a luciferase reporter assay in Cos7 cells and in human retinal pigment epithelial (ARPE-19) cells that overexpress RHO. To assess target engagement and efficacy of the oligonucleotides in vivo, we evaluatedtheir activity in mouse and pig models for RHO P23H-induced adRP. Animals received intravitreal (IVT) injections, and eyes were evaluated for RHO P23H expression and expression of rod and cone cell markers by immunohistochemistry.

Results : We demonstrate that stereopure oligonucleotides selectively deplete expression of RHO P23H transcripts in vitro. After a single IVT eye injection of a stereopure oligonucleotide, expression of RHO P23H transcripts are decreased by at least 50% in mouse and pig models for RHO P23H-induced adRP. In the pig model, a single IVT injection led to the retention of rod outer segments and cone pedicles compared with untreated eyes 16-weeks post-injection.

Conclusions : These results confirm that stereopure oligonucleotides can selectively deplete expression of RHO P23H transcripts. These oligonucleotides deplete RHOP23H transcripts in vivo in multiple models and rescue phenotypes associated with adRP. Together, these data suggest stereopure oligonucleotides may provide a viable therapeutic opportunity for addressing RHO P23H-dependent adRP.

This is a 2021 ARVO Annual Meeting abstract.