Stereopure oligonucleotides achieve allele-selective reduction of P23H mutant rhodopsin

Abstract

Purpose : PRISM^TM is Wave Life Sciences’ proprietary discovery and drug development platform that enables us to generate stereopure oligonucleotides targeting genetically defined diseases. PRISM combines our unique ability to construct stereopure oligonucleotides with a growing knowledge of how the interplay among oligonucleotide sequence, chemistry and backbone stereochemistry impacts key pharmacologic properties. Here, we report the application of PRISM to generate a stereopure oligonucleotide that promotes RNase H-mediated allele-selective degradation of mutant rhodopsin (RHO) transcripts, which encode a toxic gain-of-function protein (RHO P23H) and causes autosomal dominant retinitis pigmentosa (adRP).

Methods : Stereopure oligonucleotides were generated using PRISM. For all experiments, RNA levels were quantified by qPCR. We evaluated allele-selective activity against RHO alleles with a luciferase reporter assay in Cos7 cells. Potencies for stereopure oligonucleotides targeting RHOP23H were also evaluated in vitro in human retinal pigment epithelial (ARPE-19) cells overexpressing RHO. To assess target engagement, we quantified MALAT1, a proof of concept RNA, or RHOP23H transcripts in cellular and animal models. Animals received stereopure oligonucleotides or PBS as bilateral intravitreal (IVT) injections.

Results : Using MALAT1, we demonstrate that a single IVT injection of stereopure oligonucleotide leads to 50% knockdown in the posterior mouse eye for 9 months and >90% knockdown in the non-human primate (NHP) retina for at least 4 months. We also demonstrate that our stereopure oligonucleotide targeting RHO yields allele-selective degradation of RHOP23H RNA in biochemical and cellular experiments. Finally, we demonstrate target engagement in animal models of RHO P23H-induced adRP.

Conclusions : These results demonstrate that stereopure oligonucleotides induce potent and durable knockdown of MALAT1 in the mouse and NHP eye. We also show that stereopure oligonucleotides can achieve allele-selective knockdown of RHOP23H, suggesting that they may provide a viable therapeutic opportunity for addressing RHO P23H-dependent adRP.

This is a 2020 ARVO Annual Meeting abstract.