Generation and Characterization of Amyotrophic Lateral Sclerosis iPS cells

描述(申请人提供)：尽管我们不完全知道培养皿中细胞的疾病研究是否能完全模拟人类成年神经退行性疾病如ALS的发育进程，但新描述的从ALS患者中产生诱导多能细胞(IPS)的技术能力为我们提供了一个特殊的工具，通过它我们可以探索这些问题。最近对肌萎缩侧索硬化症的病理生理学的许多见解来自于对这种疾病的家族性形式的研究。真正拥有人类细胞系的能力--在最相关的细胞类型--运动神经元和星形胶质细胞中代表自然疾病--将提供前所未有的工具，用于1)研究导致疾病病理生理学的细胞与细胞之间的相互作用，2)为药物发现和遗传路径分析提供关键工具。最终，这些ALS细胞系也将有助于比较ALS和其他神经退行性iPS模型之间常见和不常见的通路。但是-iPS细胞生物学是非常新的，我们还没有足够的信息关于所产生的细胞的可靠性，它们是否能够真实地反映人类细胞生物学，概括天然运动神经元和星形胶质细胞的蛋白质、遗传和功能特征。在我们开始广泛使用这些细胞进行基础/翻译研究之前，关键是要产生一系列细胞系，这些细胞系都是在相同的条件下从不同的FAL突变中产生的，以确定它们在细胞类型特异性和功能生物学方面的代表性。整个提案将涉及四名主要研究人员，密切合作，以产生和评估家族性ALS(FALS)iPS细胞系。由Eggan博士领导的项目1将从FALS和对照患者那里获得皮肤活检，产生成纤维细胞，并最终产生最初的iPS系。我们将利用出水公司，一家生物技术公司，作为统一方案iPS细胞产生的中心站点。具有神经/神经胶质特征的iPS细胞系将被送往由Chris Henderson领导的项目2实验室-运动神经元生物学，以及由Jeffrey Rothstein领导的项目3实验室-星形胶质细胞。这两个项目/实验室将通过一系列顺序分析，确定哪些FAL iPS细胞系具有运动神经元和星形胶质细胞的适当特征。只有那些符合最终标准的细胞系(与人类ES细胞和之前对人类星形胶质细胞的研究相比)才会在Tom Maniatis领导的Project 4实验室进行最终的基因分析。 公共卫生相关性：了解ALS的病理生理学和新疗法的开发一直是一个巨大的挑战。真正拥有人类细胞系的能力--在最相关的细胞类型--运动神经元和星形胶质细胞中代表自然疾病--将提供前所未有的工具，用于1)研究导致疾病病理生理学的细胞与细胞之间的相互作用，2)为药物发现和遗传路径分析提供关键工具。最终，这些ALS细胞系也将有助于比较ALS和其他神经退行性iPS模型之间常见和不常见的通路。)

DESCRIPTION (provided by applicant): Although we do not fully know if disease study of cells in Petri dishes can fully emulate the developmental progression that occurs in human adult neurodegenerative disease like ALS, new described technical ability to generate Induced Pluripotent Cells (iPS) from ALS patients provides an exceptional tool by which we can explore these issues. Many recent insights into the pathophysiology of ALS come from the study of familial forms of this disease. The ability to actually have human cell lines- representing the natural disease in the most relevant cell types- motor neurons and astrocytes- will provide unprecedented tools to 1) study cell- cell interactions responsible for disease pathophysiology and 2) provide critical tools for drug discovery and genetic pathway analysis. Eventually these ALS cell lines will also be useful to compare common and uncommon pathways between ALS and other neurodegenerative iPS models. But - iPS cell biology is exceptionally new and we do not yet have sufficient information about the reliability of the cells generated, their ability to truly reflect human cell biology, recapitulate the protein, genetic and functional characteristics of native motor neurons and astroglia. Before we can embark on extensive use of these cells for basic/translational research- it would be critical to generate a series of cell lines- all produced under identical conditions, from different fALS mutations, to determine how representative they are for cell type specificity and functional biology. The overall proposal will involve four principal investigators, working in tight collaboration, to generate and evaluate familial ALS (fALS) iPS cell lines. Project 1, led by Dr. Eggan will obtain the skin biopsies from FALS and control patients, generate the fibroblast and ultimately the initial iPS lines. We will employ the aid of iZumi, a biotech company to be a central site for uniform protocol iPS cell generation. iPS cell lines with neural/glial characteristics will be sent to the Project 2 Lab- Motor neuron biology, lead by Chris Henderson and to Project 3 lab, Astrocytes- lead by Jeffrey Rothstein. These two projects/labs will determine which of the fALS iPS cell lines have the appropriate characteristics of motor neurons and astroglia, through a series of sequential analyses. Only those cell lines that meet final criteria (as compared to human ES cell and prior work on human astroglia) will then go on for final genetic analysis in the Project 4 lab, lead by Tom Maniatis. PUBLIC HEALTH RELEVANCE: Understanding the pathophysiology and development of new therapeutics for ALS has been an enormous challenge. The ability to actually have human cell lines- representing the natural disease in the most relevant cell types- motor neurons and astrocytes- will provide unprecedented tools to 1) study cell- cell interactions responsible for disease pathophysiology and 2) provide critical tools for drug discovery and genetic pathway analysis. Eventually these ALS cell lines will also be useful to compare common and uncommon pathways between ALS and other neurodegenerative iPS models. )