Use of induced pluripotent stem cells to study mechanims of familial and sporadic

摘要 ALS是一种进行性神经退行性疾病，其特征在于上皮层和下皮层的丧失。 下运动神经元，最终导致肌肉萎缩和呼吸衰竭死亡 [1-3]中所述。大多数ALS病例是散发性的，90%的患者表现为疾病 没有ALS家族史的症状。其余10%的ALS患者 被诊断为家族性ALS [1-3]。大约25%的ALS家族病例是 由显性遗传的超氧化物歧化酶基因突变引起 (SOD)[4]的文件。SOD 1致病等位基因的鉴定导致了 用于ALS研究的转基因小鼠和大鼠模型[5-8]。生产过剩 小鼠和大鼠中的致病性人SOD 1蛋白导致迟发性、进行性 神经退行性疾病[5，6，8]。对SOD 1动物模型的研究导致了 肌萎缩侧索硬化症运动神经元内在致病特性鉴定与研究 包括蛋白质聚集体的形成、细胞骨架异常、蛋白酶体 功能障碍和对细胞死亡信号的敏感性增加[1，5]。虽然很多 在这些家族性疾病的动物模型中了解到， 由于缺乏合适的体内系统而导致散发性疾病。我们建议 利用干细胞研究的最新进展[9-13]来产生和研究人类 运动神经元来源于散发性ALS患者捐赠的成纤维细胞。 这些运动神经元将用于长期细胞培养， 散发性疾病的病理生理学特征，包括蛋白质分析 聚集体，形态改变的线粒体，电生理特性，基因 表达分析和检查前体mRNA可变剪接的模式。如果 成功的，这些研究可能会提供重要的新的机制见解散发 ALS，并提供基于细胞的药物筛选试验。

Abstract ALS is a progressive neurodegenerative disease characterized by the loss of upper and lower motor neurons, culminating in muscle wasting and death from respiratory failure [1-3]. The majority of ALS cases are sporadic, with 90% of patients presenting disease symptoms with no family history of ALS. The remaining 10% of ALS patients are diagnosed with familial ALS [1-3]. Approximately 25% of the familial cases of ALS are caused by mutations in the dominantly inherited gene encoding super oxide dismutase (SOD) [4]. Identification of pathogenic alleles of SOD1 has led to the production of transgenic mouse and rat models for the study of ALS [5-8]. Overproduction of pathogenic human SOD1 protein in mice and rats leads to late onset, progressive neurodegenerative disease [5, 6, 8]. Studies of the SOD1 animal models have led to the identification and study of intrinsic pathogenic characteristics of ALS motor neurons including the formation of protein aggregates, cytoskeletal abnormalities, proteasome dysfunction and increased sensitivity to cell death signals [1, 5]. Although much has been learned in these animal models of the familial disease, very little is known about the sporadic disease because of the lack of a suitable in vivo system. We propose to make use of recent advances in stem cell research [9-13] to generate and study human motor neurons in culture derived from fibroblasts donated by sporadic ALS patients. These motor neurons will be used in long term cell cultures to examine pathophysiological hallmarks of the sporadic disease, including analysis of protein aggregates, morphologically altered mitochondria, electrophysiological properties, gene expression analyses, and examination of patterns of alternative pre-mRNA splicing. If successful, these studies may provide important new mechanistic insights into sporadic ALS, and provide cell-based assays for drug screening.