The fused in sarcoma protein forms cytoplasmic aggregates in motor neurons derived from integration-free induced pluripotent stem cells generated from a patient with familial amyotrophic lateral sclerosis carrying the FUS-P525L mutation
The fused in sarcoma protein forms cytoplasmic aggregates in motor neurons derived from integration-free induced pluripotent stem cells generated from a patient with familial amyotrophic lateral sclerosis carrying the FUS-P525L mutation
复制标题
肉瘤蛋白中的融合蛋白在运动神经元中形成细胞质聚集体，这些运动神经元源自携带 FUS-P525L 突变的家族性肌萎缩侧索硬化症患者产生的无整合诱导多能干细胞
DOI：
10.1007/s10048-015-0448-y
复制
发表时间：
2015-07-01
期刊：
影响因子：
2.2
通讯作者：
Deng, Min
中科院分区：
文献类型：
--
作者：
Liu, Xinxiu;Chen, Jiayu;Deng, Min
肌萎缩侧索硬化症（ALS）是一种致命的神经退行性疾病，主要影响运动神经元（MNs），目前尚无有效的治疗方法。融合肉瘤（FUS）基因突变和FUS蛋白异常聚集已在ALS中报道。然而，ALS的发病机制尚不清楚。由于缺乏适当的疾病模型，包括无法获得ALS患者的MNs，临床药物试验失败了。来自ALS患者的诱导多能干细胞（iPS）为体外机制研究和未来患者特异性细胞治疗提供了不可或缺的资源。先前的报告表明，从白种人群体的成纤维细胞中产生的基于病毒的ALS-iPS细胞是基础研究的理想选择；然而，ALS-iPS细胞被排除在基于细胞的治疗应用之外，因为将病毒序列整合到基因组中存在风险，并且与皮肤活检相关的不便。为了建立用于临床应用的模型，我们使用episomal载体，从携带FUS-P525L突变的家族性ALS （FALS）患者和健康对照的外周血单个核细胞（PBMCs）中获得了一株无整合的iPS细胞系。此外，我们成功地将ALS患者特异性iPS细胞分化为MNs，并随后检测到由于FUS- p525l突变导致的细胞质错定位和MNs中FUS蛋白聚集体的形成。我们的发现为进一步阐明ALS的发病机制提供了一个基于细胞的疾病模型，并为探索基因修复与细胞替代疗法的结合提供了一个工具。
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that primarily affects motor neurons (MNs) and has no effective treatment. Mutations in the fused in sarcoma (FUS) gene and abnormal aggregation of FUS protein have been reported in ALS. However, the mechanisms involved in ALS are poorly understood. Clinical drug trails have failed due to a lack of appropriate disease models, including a lack of access to MNs from ALS patients. Induced pluripotent stem (iPS) cells derived from patients with ALS provide an indispensable resource for in vitro mechanistic studies and for future patient-specific cell-based therapies. Previous reports demonstrated that viral-based ALS-iPS cells generated from fibroblasts harvested from Caucasian populations are ideal for basic research; however, ALS-iPS cells are precluded from cell-based therapeutic applications because of the risks associated with the integration of viral sequences into the genome and inconvenience associated with dermal biopsies. To establish a model for use in clinical applications, using episomal vectors, we generated an integration-free iPS cell line from peripheral blood mononuclear cells (PBMCs) harvested from a familial ALS (FALS) patient carrying the FUS-P525L mutation and a healthy control. Furthermore, we successfully differentiated ALS patient-specific iPS cells into MNs and subsequently detected cytoplasmic mislocalization and formation of FUS protein aggregates in MNs due to the FUS-P525L mutation. Our findings offer a cell-based disease model for use in further elucidating ALS pathogenesis and provide a tool for exploring gene repair coupled with cell replacement therapy.