Artificial miRNAs Reduce Human Mutant Huntingtin Throughout the Striatum in a Transgenic Sheep Model of Huntington's Disease.
Artificial miRNAs Reduce Human Mutant Huntingtin Throughout the Striatum in a Transgenic Sheep Model of Huntington's Disease.
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DOI：
10.1089/hum.2017.199
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发表时间：
2017-12
影响因子：
4.2
通讯作者：
E. Pfister;Natalie DiNardo;E. Mondo;F. Borel;Faith Conroy;Cara K. Fraser;Gwladys Gernoux;Xin Han;Danjing Hu;Emily S. Johnson;Emily S. Johnson;Lori A. Kennington;Pengpeng Liu;S. Reid;E. Sapp;P. Vodička;T. Kuchel;A. Morton;D. Howland;R. Moser;M. Sena-Esteves;G. Gao;Christian Mueller;M. Difiglia;N. Aronin
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文献类型：
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作者：
E. Pfister;Natalie DiNardo;E. Mondo;F. Borel;Faith Conroy;Cara K. Fraser;Gwladys Gernoux;Xin Han;Danjing Hu;Emily S. Johnson;Emily S. Johnson;Lori A. Kennington;Pengpeng Liu;S. Reid;E. Sapp;P. Vodička;T. Kuchel;A. Morton;D. Howland;R. Moser;M. Sena-Esteves;G. Gao;Christian Mueller;M. Difiglia;N. Aronin
亨廷顿病（HD）是由亨廷顿蛋白（HTT）基因中CAG重复区的遗传扩增引起的致命性神经退行性疾病。在HD小鼠模型中的研究表明，人工miRNA可以减少突变HTT，但缺乏在大型动物中有效性和安全性的证据。HD转基因绵羊表达具有73个CAG重复的全长人HTT。使用AAV 9将靶向人HTT mRNA外显子48的人工miRNA单侧递送至HD绵羊纹状体，所述人HTT mRNA外显子48在两个替代启动子U6或CβA的控制下。在注射后1个月和6个月，该处理使纹状体中的人突变体（m）HTT mRNA和蛋白质减少50-80%。在尾状核和壳核中都可以检测到沉默。内源性绵羊HTT蛋白水平不受影响。在治疗后6个月，DARPP 32或NeuN标记的神经元没有显著损失，在对照水平检测到Iba 1阳性小胶质细胞。结论是，通过直接递送携带人工miRNA的AAV，可以在大型动物脑中实现并维持人mHTT蛋白的安全有效沉默。
Huntington's disease (HD) is a fatal neurodegenerative disease caused by a genetic expansion of the CAG repeat region in the huntingtin (HTT) gene. Studies in HD mouse models have shown that artificial miRNAs can reduce mutant HTT, but evidence for their effectiveness and safety in larger animals is lacking. HD transgenic sheep express the full-length human HTT with 73 CAG repeats. AAV9 was used to deliver unilaterally to HD sheep striatum an artificial miRNA targeting exon 48 of the human HTT mRNA under control of two alternative promoters: U6 or CβA. The treatment reduced human mutant (m) HTT mRNA and protein 50-80% in the striatum at 1 and 6 months post injection. Silencing was detectable in both the caudate and putamen. Levels of endogenous sheep HTT protein were not affected. There was no significant loss of neurons labeled by DARPP32 or NeuN at 6 months after treatment, and Iba1-positive microglia were detected at control levels. It is concluded that safe and effective silencing of human mHTT protein can be achieved and sustained in a large-animal brain by direct delivery of an AAV carrying an artificial miRNA.