Manning Kassie S, Rao Ashish N, Castro Miguel, Cooper Thomas A
Bio-Synthesis, Inc. , 612 East Main Street, Lewisville, Texas 75057, United States.
ACS Chem Biol. 2017 Oct 20;12(10):2503-2509. doi: 10.1021/acschembio.7b00416. Epub 2017 Sep 5.
Myotonic dystrophy type 1 (DM1) is a multisystemic disease caused by an expanded CTG repeat in the 3' UTR of the dystrophia myotonica protein kinase (DMPK) gene. Short, DNA-based antisense oligonucleotides termed gapmers are a promising strategy to degrade toxic CUG expanded repeat (CUG) RNA. Nucleoside analogs are incorporated to increase gapmer affinity and stability; however, some analogs also exhibit toxicity. In this study, we demonstrate that the 2',4'-BNA[NMe] (BNA) modification is a promising nucleoside analog with high potency similar to 2',4'-LNA (LNA). BNA gapmers targeting a nonrepetitive region of the DMPK 3' UTR show allele-specific knockdown of CUG RNA and revert characteristic DM1 molecular defects including mis-splicing and accumulation of RNA foci. Notably, the BNA gapmers tested in this study did not induce caspase activation, in contrast to a sequence matched LNA gapmer. This study indicates that BNA gapmers warrant further study as a promising RNA targeting therapeutic.
1型强直性肌营养不良症（DM1）是一种多系统疾病，由肌强直性营养不良蛋白激酶（DMPK）基因3'非翻译区的CTG重复序列扩增引起。短的、基于DNA的反义寡核苷酸（称为gapmer）是降解有毒CUG扩增重复序列（CUG）RNA的一种有前景的策略。掺入核苷类似物以增加gapmer的亲和力和稳定性；然而，一些类似物也表现出毒性。在本研究中，我们证明2'，4'-BNA[NMe]（BNA）修饰是一种有前景的核苷类似物，其效力与2'，4'-锁核酸（LNA）相似。靶向DMPK 3'非翻译区非重复区域的BNA gapmer显示CUG RNA的等位基因特异性敲低，并逆转包括剪接错误和RNA病灶积累在内的特征性DM1分子缺陷。值得注意的是，与序列匹配的LNA gapmer相比，本研究中测试的BNA gapmer未诱导半胱天冬酶激活。本研究表明，BNA gapmer作为一种有前景的RNA靶向治疗药物值得进一步研究。
