Song Guoxu, Ma Yuying, Gao Xing, Zhang Xuewen, Zhang Fei, Tian Chunhong, Hou Jiajia, Liu Zheng, Zhao Zixin, Tian Yong
Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Life Med. 2022 Jun 29;1(1):27-44. doi: 10.1093/lifemedi/lnac020. eCollection 2022 Aug.
The neurodegenerative disease spinocerebellar ataxia type 3 (SCA3; also called Machado-Joseph disease, MJD) is a trinucleotide repeat disorder caused by expansion of the CAG repeats in the gene. Here, we applied a CRISPR/Cas9-mediated approach using homologous recombination to achieve a one-step genetic correction in SCA3-specific induced pluripotent stem cells (iPSCs). The genetic correction reversed disease-associated phenotypes during cerebellar region-specific differentiation. In addition, we observed spontaneous ataxin-3 aggregates specifically in mature cerebellar neurons differentiated from SCA3 iPSCs rather than in SCA3 pan-neurons, SCA3 iPSCs or neural stem cells, suggesting that SCA3 iPSC-derived disease-specific and region-specific cerebellar neurons can provide unique cellular models for studying SCA3 pathogenesis . Importantly, the genetically corrected cerebellar neurons did not display typical SCA3 aggregates, suggesting that genetic correction can subsequently reverse SCA3 disease progression. Our strategy can be applied to other trinucleotide repeat disorders to facilitate disease modeling, mechanistic studies and drug discovery.
神经退行性疾病3型脊髓小脑共济失调（SCA3；也称为马查多-约瑟夫病，MJD）是一种由该基因中CAG重复序列扩增引起的三核苷酸重复序列疾病。在此，我们应用了一种基于同源重组的CRISPR/Cas9介导方法，在SCA3特异性诱导多能干细胞（iPSC）中实现一步基因校正。这种基因校正逆转了小脑区域特异性分化过程中与疾病相关的表型。此外，我们观察到ataxin-3聚集体自发地特异性出现在由SCA3 iPSC分化而来的成熟小脑神经元中，而非SCA3全神经元、SCA3 iPSC或神经干细胞中，这表明SCA3 iPSC来源的疾病特异性和区域特异性小脑神经元可为研究SCA3发病机制提供独特的细胞模型。重要的是，经基因校正的小脑神经元未显示出典型的SCA3聚集体，这表明基因校正随后可逆转SCA3疾病进展。我们的策略可应用于其他三核苷酸重复序列疾病，以促进疾病建模、机制研究和药物发现。
