Optimization of trans-Splicing for Huntington's Disease RNA Therapy.
Optimization of trans-Splicing for Huntington's Disease RNA Therapy.
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DOI：
10.3389/fnins.2017.00544
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发表时间：
2017
影响因子：
4.3
通讯作者：
Mattis VB
中科院分区：
文献类型：
--
作者：
Rindt H;Tom CM;Lorson CL;Mattis VB
亨廷顿病 (HD) 是一种毁灭性的神经退行性疾病，由亨廷顿 (HTT) 基因外显子 1 的聚谷氨酰胺 (polyQ) 扩增引起。我们之前已经证明，剪接体介导的反式剪接是特异性减少和修复突变 HTT (mtHTT) 的可行分子策略。在这里，HTT 中前 mRNA 反式拼接模块 (PTM) 的靶向束缚功效得到了优化。靶向 HTT 内含子 1 3' 端或内含子 1 分支点的各种 PTM 被显示反式剪接成 HTT 小基因以及内源性 HTT 前 mRNA。专门针对内源内含子 1 分支点的 PTM 将反式剪接效率从 1-5% 提高到 10-15%。此外，人类 HD 患者 iPSC 衍生的神经培养物中 PTM 的慢病毒表达显着逆转了先前建立的两种 PolyQ 长度依赖性表型。这些结果表明，mtHTT 的前 mRNA 修复可以保持治疗效果，并且它展示了在 HD 背景下纠正 mtHTT 等位基因产生的 mRNA 产物的替代平台。
Huntington's disease (HD) is a devastating neurodegenerative disorder caused by a polyglutamine (polyQ) expansion in exon 1 of the Huntingtin (HTT) gene. We have previously demonstrated that spliceosome-mediated trans-splicing is a viable molecular strategy to specifically reduce and repair mutant HTT (mtHTT). Here, the targeted tethering efficacy of the pre-mRNA trans-splicing modules (PTM) in HTT was optimized. Various PTMs that targeted the 3′ end of HTT intron 1 or the intron 1 branch point were shown trans-splice into an HTT mini-gene, as well as the endogenous HTT pre-mRNA. PTMs that specifically target the endogenous intron 1 branch point increased the trans-splicing efficacy from 1–5 to 10–15%. Furthermore, lentiviral expression of PTMs in a human HD patient iPSC-derived neural culture significantly reversed two previously established polyQ-length dependent phenotypes. These results suggest that pre-mRNA repair of mtHTT could hold therapeutic benefit and it demonstrates an alternative platform to correct the mRNA product produced by the mtHTT allele in the context of HD.
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