RNAI for Huntington's Disease

RNAI 治疗亨廷顿病

• 批准号: 6897080
• 负责人: Beverly L. Davidson
• 金额: $ 26.91万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-12-01 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6897080
• 关键词: Huntington' s disease; RNA interference; alleles; biotechnology; complementary DNA; disease /disorder model; gene delivery system; gene induction /repression; gene mutation; genetic polymorphism; genetic transduction; genetically modified animals; laboratory mouse; microarray technology; mutant; nervous system disorder therapy; neurogenetics; neuroprotectants; nonhuman therapy evaluation; therapy design /development; tissue /cell culture

This project is focused on the development of RNAi as therapy for HD. We previously demonstrated the ability of RNAi to silence a disease allele in an animal model of a related neurogenetic disease, Spinocerebellar ataxia type 1, resulting in improved behavior and neuropathology. We will now test several specific hypotheses regarding inhibition of mutant htt expression by RNAi: 1) RNAi can protect, and/or reverse, the neuropathology in mouse models of human Huntington's disease. Earlier studies in an HD mouse model with an inducible mutant allele demonstrate that if expression of the disease allele is abrogated, behavior and pathology improve. RNAi for silencing could yield a similar, promising benefit. 2) RNAi targeted to htt cDNA polymorphisms can be utilized for allele-specific silencing of mutant htt. One published disease-linked polymorphism and several novel ones will be tested for their utility to specifically silence the disease allele in cell culture studies. 3) Regulated RNAi can be achieved in vivo, and can be used to address duration of efficacy. HD usually takes decades to develop. If RNAi is beneficial it is unlikely that lifelong suppression of expression is required for sustained benefit. Thus we will take advantage of our recently developed regulated RNAi vectors to determine how long RNAi-induced improvements last and whether any problems from expressing RNAi in HD brain resolve once vector-expressed RNAi is reduced. 4) The off target effects of RNAi are minimal and can be resolved by cessation of RNAi. Earlier microarray analyses on HD mice brain from various models have shown that transcriptional changes induced by mutant htt are confined to a circumscribed set of genes. Regulatable vectors allow us to test if off-target effects resolve when shRNAs are no longer expressed.

该项目的重点是RNAi作为HD疗法的发展。我们先前证明了RNAi在相关神经遗传学疾病，脊髓脑性共济失调1型动物模型中静音的能力，从而改善了行为和神经病理学。现在，我们将测试有关RNAi突变HTT表达抑制突变体HTT表达的几个特定假设： 1）RNAi可以保护和/或反向人类亨廷顿氏病小鼠模型中的神经病理学。在具有诱导性突变等位基因的HD小鼠模型中的早期研究表明，如果疾病的表达被废除，行为和病理学会有所改善。用于沉默的RNAi可能会产生类似的有希望的好处。 2）针对HTT cDNA多态性的RNAi可以用于突变体HTT的等位基因特异性沉默。一种发表的疾病连接多态性和几种新颖的多态性将被测试，以使其在细胞培养研究中特别使疾病等位基因的效用进行测试。 3）可以在体内实现受调节的RNAi，可用于解决疗效持续时间。高清通常需要数十年才能发展。如果RNAi是有益的，则不太可能需要终生抑制表达才能持续受益。因此，我们将利用我们最近开发的调节的RNAi载体来确定RNAI诱导的持续时间的长时间，以及一旦减少了载体表达的RNAi，在HD脑中表达RNAi的任何问题是否降低。 4）RNAi的关闭目标效应很小，可以通过停止RNAi解决。对来自各种模型的HD小鼠大脑的早期微阵列分析表明，突变体HTT诱导的转录变化仅限于一组受限制的基因集。可调节的向量使我们能够测试不再表达shRNA时脱离靶向效应的解决方案。