Enhanced splicing modulation by NMA-modified antisense oligonucleotides

Abstract

Aberrant RNA splicing contributes to many human diseases, and splice-switching antisense oligonucleotides (SSOs) are ideally suited as a therapeutic strategy to modulate splicing and restore normal gene expression. Nusinersen (Spinraza) has revolutionized the treatment of spinal muscular atrophy. It is a splice-switching oligonucleotide (SSO) that is modified with 2-O-methoxyethyl (MOE) modifications. Here, we introduce a next-generation ribose modification, 2'-O-[2-(methyl-amino)-2-oxoethyl] (NMA), which enhances the pharmacological properties of SSOs. We identified a long-lasting NMA-modified human candidate SSO, salanersen, that is 3-4-fold more potent than nusinersen in human SMN2 transgenic mice. To evaluate the generality of the NMA chemistry, we applied it to modulation of SCN1A exon 20N splicing, a therapeutic strategy for Dravet syndrome. An NMA-modified SSO is 3.5-fold more potent than STK-001, a MOE-modified SSO currently in clinical trials. Our data establish the NMA chemistry as a broadly applicable ribose modification that markedly improves the pharmacological profile of SSOs, supporting its development as a next-generation platform for splicing modulation therapies. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=69 SRC="FIGDIR/small/680653v1_ufig1.gif" ALT="Figure 1"> View larger version (16K): org.highwire.dtl.DTLVardef@122e426org.highwire.dtl.DTLVardef@1b0a56eorg.highwire.dtl.DTLVardef@3ce4e8org.highwire.dtl.DTLVardef@1d86378_HPS_FORMAT_FIGEXP M_FIG C_FIG