Generation and Characterization of Amyotrophic Lateral Sclerosis

肌萎缩侧索硬化症的产生和特征

• 批准号: 8488511
• 负责人: KEVIN C EGGAN
• 金额: $ 79万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8488511
• 关键词: Amyotrophic Lateral Sclerosis; Applications Grants; Astrocytes; Biological Assay; Biology; Cell Communication; Cell Line; Cells; Characteristics; Collaborations; Collection; Complement; Data; Development; Disease; Ensure; Evaluation; Fibroblasts; Functional disorder; Future; Generations; Genes; Genetic; Genotype; Grant; Human; Human Cell Line; In Vitro; Individual; Informatics; Lead; Life; Modeling; Monitor; Motor Neurons; Mutation; Pathogenesis; Pathway Analysis; Patients; Preclinical Drug Evaluation; Principal Investigator; Process; Protocols documentation; Quality Control; Reporter; Research; Research Personnel; Specificity; System; Therapeutic; Validation; Work; Zinc Fingers; assay development; base; cell type; deep sequencing; disease phenotype; drug discovery; genetic analysis; high standard; human embryonic stem cell; in vivo; induced pluripotent stem cell; insight; meetings; new therapeutic target; novel; novel therapeutics; pluripotency; relating to nervous system; screening; tool; tool development

Induced Pluripotent Cells (IPS) from ALS patients could provide an exceptional tool by which we can Disease pathophysiology and druggable targets. 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- provides unprecedented tools to 1) study cell interactions responsible for disease pathophysiology and 2) provide critical tools for drug discovery and genetic pathway analysis. We have generated iPS cell lines, under identical conditions, from various familial ALS (fALS) mutations and sporadic ALS (sALS) patients to determine how representative they are for cell type specificity and functional biology. The first set of cells now generated are ready for use in disease phenotyping paradigms. The overall proposal will involve four principal investigators, working in tight collaboration, to generate and evaluate fALS and sALS cell lines and to use them to develop cell specific phenotypic assays. Project 1, lead by Dr. Eggan will generate new critical IPS line- isogenic liens from selected fALS mutations as well as non-integrating IPS lines- to complement our retroviral based IPS collection. In addition Project 1 will fully evaluate the pluripotency of the lines using a novel genetic scorecard system. 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, thru sequential analyses. In addition, both groups will generate Zinc-finger based cell specific reporter cell lines for future use in drug discovery assays. Cell lines that meet final criteria (as compared to human ES cell and prior work on human astroglia) will undergo genetic analysis in the Project 4 lab, lead by Tom Maniatis. Finally, to develop useful tools for therapeutics and pathogenesis, Cores 1 and 2 will generate motor neuron and astroglial disease phenotyping assays.

来自 ALS 患者的诱导多能细胞 (IPS) 可以提供一种特殊的工具，我们可以通过它来研究疾病病理生理学和药物靶标。最近对 ALS 病理生理学的许多见解都来自对该疾病家族形式的研究。实际上拥有人类细胞系（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）的能力提供了前所未有的工具：1）研究负责疾病病理生理学的细胞相互作用，2）为药物发现和遗传途径分析提供关键工具。我们在相同条件下从各种家族性 ALS (fALS) 突变和散发性 ALS (sALS) 患者中生成了 iPS 细胞系，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。现在生成的第一组细胞已准备好用于疾病表型分析范例。总体提案将涉及四名主要研究人员，紧密合作，生成和评估 fALS 和 sALS 细胞系，并使用它们开发细胞特异性表型测定。由 Eggan 博士领导的项目 1 将产生新的关键 IPS 系——来自选定的 fALS 突变的等基因留置权以及非整合 IPS 系——以补充我们基于逆转录病毒的 IPS 系列。此外，项目 1 将使用新型遗传记分卡系统全面评估品系的多能性。具有神经/胶质细胞特征的 iPS 细胞系将​​被送往由 Chris Henderson 领导的项目 2 运动神经元生物学实验室和项目 3 实验室。星形胶质细胞——由 Jeffrey Rothstein 领导。这两个项目/实验室将通过顺序分析确定哪些 fALS IPS 细胞系具有运动神经元和星形胶质细胞的适当特征。此外，两个小组将生成基于锌指的细胞特异性报告细胞系，以供将来用于药物发现分析。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）将在由 Tom Maniatis 领导的 Project 4 实验室进行遗传分析。最后，为了开发治疗和发病机制的有用工具，核心 1 和 2 将生成运动神经元和星形胶质细胞疾病表型分析。