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Vinod Vathipadiekal, Lankai Guo, Yuan Yin, Qiong Shen, Hailin Yang, Richard Looby, Lauren Norwood, James Fransen, archana jalligampala, Jennifer Noel, Chandra Vargeese, Maureen A McCall, Michael Byrne; Stereopure oligonucleotides achieve allele-selective reduction of P23H mutant rhodopsin. Invest. Ophthalmol. Vis. Sci. 2020;61(7):1912.
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© ARVO (1962-2015); The Authors (2016-present)
PRISMTM 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).
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.
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.
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.
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