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

使用诱导多能干细胞研究家族性和散发性的机制

• 批准号: 7692283
• 负责人: THOMAS P MANIATIS
• 金额: $ 83.7万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7692283
• 关键词: Alleles; Animal Model; Biological Assay; Cell Culture Techniques; Cell Death Signaling Process; Cells; Cessation of life; Characteristics; Diagnosis; Disease; Familial disease; Family history of; Fibroblasts; Functional disorder; Gene Expression; Genes; Human; Inherited; Learning; Mitochondria; Motor Neurons; Mus; Muscle; Mutation; Neurodegenerative Disorders; Oxides; Patients; Pattern; Preclinical Drug Evaluation; Production; Property; Proteins; RNA Splicing; Rattus; Respiratory Failure; Stem Cell Research; Study models; Symptoms; System; Transgenic Mice; abstracting; base; in vivo; induced pluripotent stem cell; insight; mRNA Precursor; multicatalytic endopeptidase complex; protein aggregate; wasting

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.

抽象的 ALS是一种进行性神经退行性疾病，其特征是上层和 较低的运动神经元，最终导致肌肉浪费和呼吸衰竭死亡 [1-3]。大多数ALS病例是零星的，有90％的患者出现疾病 没有ALS家族史的症状。其余10％的ALS患者是 被诊断为家族性ALS [1-3]。大约25％的ALS病例是 由主要遗传基因编码超级氧化物歧化酶的突变引起 （SOD）[4]。 SOD1病原等位基因的鉴定导致产生 用于研究ALS的转基因小鼠和大鼠模型[5-8]。过量生产 小鼠和大鼠的致病性人类SOD1蛋白会导致发作迟到，进行性 神经退行性疾病[5，6，8]。 SOD1动物模型的研究导致了 ALS运动神经元内在致病特性的识别和研究 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]。虽然有很多 在这些家族疾病的动物模型中被学到了，对 由于缺乏合适的体内系统，这种零星疾病。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 源自零星ALS患者捐赠的成纤维细胞的培养中运动神经元。 这些运动神经元将用于长期细胞培养物检查 零星疾病的病理生理标志，包括蛋白质的分析 聚集体，形态学改变的线粒体，电生理特性，基因 表达分析和替代前MRNA剪接模式的检查。如果 成功，这些研究可能会为零星提供重要的新机械见解 ALS，并提供基于细胞的药物筛查测定法。