Establishment of In Vitro FUS-Associated Familial Amyotrophic Lateral Sclerosis Model Using Human Induced Pluripotent Stem Cells.
Establishment of In Vitro FUS-Associated Familial Amyotrophic Lateral Sclerosis Model Using Human Induced Pluripotent Stem Cells.
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DOI：
10.1016/j.stemcr.2016.02.011
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发表时间：
2016-04-12
影响因子：
5.9
通讯作者：
Okano H
中科院分区：
文献类型：
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作者：
Ichiyanagi N;Fujimori K;Yano M;Ishihara-Fujisaki C;Sone T;Akiyama T;Okada Y;Akamatsu W;Matsumoto T;Ishikawa M;Nishimoto Y;Ishihara Y;Sakuma T;Yamamoto T;Tsuiji H;Suzuki N;Warita H;Aoki M;Okano H
肌萎缩侧索硬化症(ALS)是一种晚发型运动神经元疾病。虽然其神经病理已被很好地理解，但由于目前可用的人类遗传数据的限制，其细胞和分子机制尚未阐明。在本研究中，我们通过基因组编辑技术从两名携带FUS H517D杂合突变的融合肉瘤(FUS)基因错义突变的家族性ALS(FAL)患者中获得了诱导多能干细胞(IPSC)，并利用基因组编辑技术获得了携带FUS H517D突变的等基因IPSC。这些细胞来源的运动神经元模仿了几种神经退化的表型，包括在应激条件下FUS错误定位于胞浆和应激颗粒，以及细胞脆弱性。此外，使用运动神经元前体细胞(MPC)结合CLIP-SEQ数据集的外显子阵列分析显示FALS MPC中的基因表达和/或剪接模式异常。这些结果表明，IPSC来源的运动神经元是分析人类运动神经元疾病发病机制的有用工具。利用IPSC来源的运动神经元在FAL运动神经元前体细胞中异常基因表达和/或剪接建立家族性ALS(FALS)模型突变的FUS蛋白在FAL运动神经元中的错误定位增加FAL运动神经元的凋亡Okano和Yano及其同事从两名具有FUS基因错义突变的家族性ALS患者中建立了ipSCs，并将其分化为运动神经元，作为一种新的ALS体外模型。此外，该模型能够通过患者来源的运动神经元的内源性突变来追求各种细胞生物学现象中的相关性和分子病理生理学的新发现。
Amyotrophic lateral sclerosis (ALS) is a late-onset motor neuron disorder. Although its neuropathology is well understood, the cellular and molecular mechanisms are yet to be elucidated due to limitations in the currently available human genetic data. In this study, we generated induced pluripotent stem cells (iPSC) from two familial ALS (FALS) patients with a missense mutation in the fused-in sarcoma (FUS) gene carrying the heterozygous FUS H517D mutation, and isogenic iPSCs with the homozygous FUS H517D mutation by genome editing technology. These cell-derived motor neurons mimicked several neurodegenerative phenotypes including mis-localization of FUS into cytosolic and stress granules under stress conditions, and cellular vulnerability. Moreover, exon array analysis using motor neuron precursor cells (MPCs) combined with CLIP-seq datasets revealed aberrant gene expression and/or splicing pattern in FALS MPCs. These results suggest that iPSC-derived motor neurons are a useful tool for analyzing the pathogenesis of human motor neuron disorders. Modeling familial ALS (FALS) using iPSC-derived motor neurons Aberrant gene expression and/or splicing in FALS motor neuron precursor cells Mis-localization of mutant FUS protein in FALS motor neurons Increased apoptosis of FALS motor neurons Okano, Yano, and colleagues established iPSCs from two familial ALS patients with the missense mutation in the FUS gene and differentiated them into motor neurons as a novel in vitro model for ALS. Moreover, this model enables the pursuit of correlations and new discovery of molecular pathophysiology in various cell biological phenomena by endogenous mutation in patient-derived motor neurons.