A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington's Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities.
A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington's Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities.
复制标题
DOI：
10.3390/jpm10040215
复制
发表时间：
2020-11-09
影响因子：
--
通讯作者：
Malakhova A
中科院分区：
文献类型：
--
作者：
Malankhanova T;Suldina L;Grigor'eva E;Medvedev S;Minina J;Morozova K;Kiseleva E;Zakian S;Malakhova A
亨廷顿病(HD)是一种严重的神经退行性疾病，由HTT基因第一外显子CAG三联体扩张引起。在这里，我们报道了通过CRISPR/Cas9介导的同源重组将HD突变引入健康人胚胎成纤维细胞的基因组中。我们验证了创建的HTT编辑系统的特异性，并证实在非靶点没有不必要的基因组修改。我们发现，通过对成纤维细胞克隆重新编程而获得的突变和对照等基因诱导的多能干细胞(IPSCs)都可以分化为纹状体介质中的棘神经元。接下来，我们展示了突变的IPSC来源的神经细胞的表型异常，包括神经花环的形成受损和对生长因子撤除的敏感性增加。此外，利用电子显微镜分析，我们在突变的神经元中发现了一系列的超微结构缺陷，这些缺陷不包含Huntingtin聚集体，表明这些缺陷在HD发育的早期就出现了。因此，我们的研究描述了一个新的基于IPSC的等基因细胞系统的创建，该系统模拟HD并概括突变细胞中HD特定的干扰，包括首次实现的一些超微结构特征。
Huntington’s disease (HD) is a severe neurodegenerative disorder caused by a CAG triplet expansion in the first exon of the HTT gene. Here we report the introduction of an HD mutation into the genome of healthy human embryonic fibroblasts through CRISPR/Cas9-mediated homologous recombination. We verified the specificity of the created HTT-editing system and confirmed the absence of undesirable genomic modifications at off-target sites. We showed that both mutant and control isogenic induced pluripotent stem cells (iPSCs) derived by reprogramming of the fibroblast clones can be differentiated into striatal medium spiny neurons. We next demonstrated phenotypic abnormalities in the mutant iPSC-derived neural cells, including impaired neural rosette formation and increased sensitivity to growth factor withdrawal. Moreover, using electron microscopic analysis, we detected a series of ultrastructural defects in the mutant neurons, which did not contain huntingtin aggregates, suggesting that these defects appear early in HD development. Thus, our study describes creation of a new isogenic iPSC-based cell system that models HD and recapitulates HD-specific disturbances in the mutant cells, including some ultrastructural features implemented for the first time.
登录
查看更多内容
影响因子：
25
作者：
Lo Sardo, Valentina;Zuccato, Chiara;Cattaneo, Elena
通讯作者：
Cattaneo, Elena
影响因子：
5.7
作者：
Dent EW;Merriam EB;Hu X
通讯作者：
Hu X
影响因子：
12.6
作者：
Liu Y;Zhu X
通讯作者：
Zhu X
影响因子：
5.3
作者：
Arrasate, Montserrat;Finkbeiner, Steven
通讯作者：
Finkbeiner, Steven
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle