CRISPR/Cas9 mediated gene correction ameliorates abnormal phenotypes in spinocerebellar ataxia type 3 patient-derived induced pluripotent stem cells

作者：Lang He, Shang Wang, Linliu Peng, Huifang Zhao, Shuai Li, Xiaobo Han, Jean de Dieu Habimana, Zhao Chen, Chunrong Wang, Yun Peng, Huirong Peng, Yue Xie, Lijing Lei, Qi Deng, Linlin Wan, Na Wan, Hongyu Yuan, Yiqing Gong, Guangdong Zou, Zhiyuan Li, Beisha Tang, Hong Jiang · 发表于：Translational Psychiatry · 年份：2021 · DOI：10.1038/s41398-021-01605-2 · 被引用次数：48 · 研究领域：Genetic Neurodegenerative Diseases、CRISPR and Genetic Engineering、Pluripotent Stem Cells Research

Abstract Spinocerebellar ataxia type 3/Machado–Joseph disease (SCA3/MJD) is a progressive autosomal dominant neurodegenerative disease caused by abnormal CAG repeats in the exon 10 of ATXN3 . The accumulation of the mutant ataxin-3 proteins carrying expanded polyglutamine (polyQ) leads to selective degeneration of neurons. Since the pathogenesis of SCA3 has not been fully elucidated, and no effective therapies have been identified, it is crucial to investigate the pathogenesis and seek new therapeutic strategies of SCA3. Induced pluripotent stem cells (iPSCs) can be used as the ideal cell model for the molecular pathogenesis of polyQ diseases. Abnormal CAG expansions mediated by CRISPR/Cas9 genome engineering technologies have shown promising potential for the treatment of polyQ diseases, including SCA3. In this study, SCA3-iPSCs can be corrected by the replacement of the abnormal CAG expansions (74 CAG) with normal repeats (17 CAG) using CRISPR/Cas9-mediated homologous recombination (HR) strategy. Besides, corrected SCA3-iPSCs retained pluripotent and normal karyotype, which can be differentiated into a neural stem cell (NSCs) and neuronal cells, and maintained electrophysiological characteristics. The expression of differentiation markers and electrophysiological characteristics were similar among the neuronal differentiation from normal control iPSCs (Ctrl-iPSCs), SCA3-iPSCs, and isogenic control SCA3-iPSCs. Furthermore, this study proved that the phenotypic abnormalities ...