Modulation of nuclear REST by alternative splicing: a potential therapeutic target for Huntington's disease

Abstract

Huntington's disease (HD) is caused by a genetically mutated huntingtin (mHtt) protein with expanded polyQ stretch, which impairs cytosolic sequestration of the repressor element-1 silencing transcription factor (REST), resulting in excessive nuclear REST and subsequent repression of neuronal genes. We recently demonstrated that REST undergoes extensive, context-dependent alternative splicing, of which exon-3 skipping (∆E₃ )-a common event in human and nonhuman primates-causes loss of a motif critical for REST nuclear targeting. This study aimed to determine whether ∆E₃ can be targeted to reduce nuclear REST and rescue neuronal gene expression in mouse striatal-derived, mHtt-expressing STHdh^Q111/Q111 cells-a well-established cellular model of HD. We designed two morpholino antisense oligos (ASOs) targeting the splice sites of Rest E₃ and examined their effects on ∆E₃ , nuclear Rest accumulation and Rest-controlled gene expression in STHdh^Q111/Q111 cells. We found that (1) the ASOs treatment significantly induced ∆E₃ , reduced nuclear Rest, and rescued transcription and/or mis-splicing of specific neuronal genes (e.g. Syn1 and Stmn2) in STHdh^Q111/Q111 cells; and (2) the ASOs-induced transcriptional regulation was dependent on ∆E₃ induction and mimicked by siRNA-mediated knock-down of Rest expression. Our findings demonstrate modulation of nuclear REST by ∆E₃ and its potential as a new therapeutic target for HD and provide new insights into environmental regulation of genome function and pathogenesis of HD. As ∆E₃ is modulated by cellular signalling and linked to various types of cancer, we anticipate that ∆E₃ contributes to environmentally tuned REST function and may have a broad range of clinical implications.

Keywords: Huntington's disease; PPARγ; REST/NRSF; Stmn2; Syn-1; alternative splicing; antisense oligos; gene therapy; nuclear translocation.