Potent and Selective Antisense Oligonucleotides Targeting Single-Nucleotide Polymorphisms in the Huntington Disease Gene/Allele-Specific Silencing of Mutant Huntingtin
Potent and Selective Antisense Oligonucleotides Targeting Single-Nucleotide Polymorphisms in the Huntington Disease Gene/Allele-Specific Silencing of Mutant Huntingtin
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DOI：
10.1038/mt.2011.201
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发表时间：
2011-12-01
影响因子：
12.4
通讯作者：
Hayden, Michael R.
中科院分区：
文献类型：
--
作者：
Carroll, Jeffrey B.;Warby, Simon C.;Hayden, Michael R.
亨廷顿病 (HD) 是一种常染色体显性神经退行性疾病，由亨廷顿基因 (HTT) 中的 CAG 扩展引起，导致突变型亨廷顿蛋白 (mHTT) 的功能出现毒性增强。因此，减少 mHTT 的表达是治疗 HD 的一种有吸引力的疗法。然而，野生型 HTT 蛋白对于发育至关重要，并且在维持神经元健康方面发挥着关键作用。因此，减少野生型 HTT 的 HD 疗法可能会产生意想不到的负面后果。我们已经在人类 HD 人群中确定了单核苷酸多态性 (SNP) 靶点，用于 HTT 基因的疾病特异性靶向。使用 HD 患者的原代细胞以及转基因 YAC18 和 BACHD 小鼠系，我们开发了反义寡核苷酸 (ASO) 分子，可有效且选择性地沉默外显子和内含子 SNP 位点的 mHTT。用 S-约束乙基 (cET) 基序对这些 ASO 进行修饰可显着提高效力，同时在体外选择性地保持等位基因。所开发的 ASO 在递送至小鼠大脑后对体内 mHTT 具有有效和选择性。我们证明，在体外和体内使用 ASO 可以在治疗相关的 SNP 位点实现 mHTT 蛋白的有效和选择性等位基因特异性敲低。 2011 年 5 月 25 日收稿； 2011 年 8 月 24 日接受； 2011 年 10 月 4 日在线发布。doi: 10.1038/mt.2011.201
Huntington disease (HD) is an autosomal dominant neurodegenerative disorder caused by CAG-expansion in the huntingtin gene (HTT) that results in a toxic gain of function in the mutant huntingtin protein (mHTT). Reducing the expression of mHTT is therefore an attractive therapy for HD. However, wild-type HTT protein is essential for development and has critical roles in maintaining neuronal health. Therapies for HD that reduce wild-type HTT may therefore generate unintended negative consequences. We have identified single-nucleotide polymorphism (SNP) targets in the human HD population for the disease-specific targeting of the HTT gene. Using primary cells from patients with HD and the transgenic YAC18 and BACHD mouse lines, we developed antisense oligonucleotide (ASO) molecules that potently and selectively silence mHTT at both exonic and intronic SNP sites. Modification of these ASOs with S-constrainedethyl (cET) motifs significantly improves potency while maintaining allele selectively in vitro. The developed ASO is potent and selective for mHTT in vivo after delivery to the mouse brain. We demonstrate that potent and selective allele-specific knockdown of the mHTT protein can be achieved at therapeutically relevant SNP sites using ASOs in vitro and in vivo. Received 25 May 2011; accepted 24 August 2011; published online 04 October 2011. doi: 10.1038/mt.2011.201