Generation and Characterization of Amyotrophic Lateral Sclerosis iPS cells

描述（由申请人提供）：尽管我们不完全知道培养皿中细胞的疾病研究是否可以完全模拟人类成人神经退行性疾病（如ALS）的发育过程，但新描述的从ALS患者中产生诱导多能细胞（iPS）的技术能力为我们探索这些问题提供了一个特殊的工具。最近对ALS病理生理学的许多见解来自对这种疾病的家族形式的研究。实际拥有人类细胞系的能力——在最相关的细胞类型中代表自然疾病——运动神经元和星形胶质细胞——将提供前所未有的工具：1)研究导致疾病病理生理学的细胞-细胞相互作用；2)为药物发现和遗传途径分析提供关键工具。最终，这些肌萎缩侧索硬化症细胞系也将用于比较肌萎缩侧索硬化症和其他神经退行性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，关于产生的细胞的可靠性，他们真正反映人类细胞生物学的能力，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。在我们开始将这些细胞广泛用于基础/转化研究之前，至关重要的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fALS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。总体提案将涉及四位主要研究人员，他们密切合作，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患者和对照患者那里获得皮肤活检，生成成纤维细胞并最终生成初始iPS细胞系。我们将利用生物技术公司iZumi的帮助，成为统一协议iPS细胞生成的中心站点。”具有神经/胶质特征的iPS细胞系将被送到由克里斯·亨德森（Chris Henderson）领导的项目2实验室——运动神经元生物学，以及由杰弗里·罗斯坦（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的序列分析，确定哪些fALS 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（与人类胚胎干细胞和先前在人类星形胶质细胞上的研究相比）才会在Tom Maniatis领导的Project 4实验室进行最终的基因分析。

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. )