Regulation of Stem Cell Development

干细胞发育的调控

• 批准号: 7676633
• 负责人: IRWIN D BERNSTEIN
• 金额: $ 4.4万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-15 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7676633
• 关键词: Address; Adult; Affect; Biological Process; CD34 gene; Cells; Collaborations; Development; Embryo; Epigenetic Process; Evolution; Fred Hutchinson Cancer Research Center; Gene Expression; Generations; Genome; Goals; Heart; Hematopoietic; Hematopoietic stem cells; Human; In Vitro; Ligands; Lung; Methods; Modeling; Molecular Profiling; Mutation; Pathway interactions; Population; RNA Interference; Reagent; Regulation; Research Design; Signal Transduction; Stem Cell Development; Stem cells; Technology; Testing; Therapeutic; Umbilical Cord Blood; Universities; Washington; base; cell type; clinical application; experience; in vivo; induced pluripotent stem cell; member; multidisciplinary; notch protein; progenitor; self-renewal; stem; stem cell biology

DESCRIPTION (provided by applicant): Therapeutic application of cells derived from embryonic stem (ES) or "reprogrammed" stem cells may require, for certain cells types such as hematopoietic stem cells (HSC), expansion of these populations without introducing genetic alterations to induce self-renewal. To address this issue, we propose to test our hypotheses that pathways that regulate normal development can be manipulated to generate expanded populations of cell types that reflect normal, developmental states. As a model for this approach, we focus on the Notch and Wnt pathways with which we have extensive experience. Thus, ES and induced pluripotent stem (IPS) cells will be assessed for development of enhanced numbers of HSC by manipulation of the Notch (project 1) and Wnt (project 2) pathways. As part of each of these projects, global assessment of gene expression and epigenetic state is planned to respectively determine key similarities or differences between in vitro derived hematopoietic cells and isolated normal counterparts. Based on our current generation and clinical application of human CD34+ stem/progenitor cells using the Notch ligand Delta1, we will also test the in vivo biological function of a well characterized in vitro generated stem/progenitor population and assess the significance of potential deviations from "normal" molecular profiles (Project 3). To further understand mechanisms involved in Notch- and Wnt-induced affects on self-renewal and differentiation, we will perform genome-scale, "multiplexed" RNAi screens in expanding populations of ES/IPS and cord blood derived CD34+ cells (project 4). The assembled team represents multidisciplinary expertise at the Fred Hutchinson Cancer Research Center (FHCRC) and the University of Washington (UW) in hematopoietic stem cell biology, Wnt and Notch signaling, epigenetics, evolution of gene expression in development, genome-scale RNAi technology, and requisite experience in ES and IPS. We anticipate providing the expertise, as well as unique reagents for modulating Notch and Wnt signaling, in collaborations with other members of the stem cell consortium evaluating heart and lung as well as hematopoietic progenitors.

描述（由申请人提供）：对于某些细胞类型如造血干细胞（HSC），来自胚胎干（ES）或“重编程”干细胞的细胞的治疗应用可能需要扩增这些群体而不引入遗传改变以诱导自我更新。为了解决这个问题，我们建议测试我们的假设，即调节正常发育的途径可以被操纵，以产生反映正常发育状态的细胞类型的扩增群体。作为这种方法的模型，我们专注于Notch和Wnt途径，我们拥有丰富的经验。因此，将通过操作Notch（项目1）和Wnt（项目2）途径评估ES和诱导多能干细胞（IPS）的HSC数量增加。作为这些项目的一部分，计划对基因表达和表观遗传状态进行全球评估，以分别确定体外衍生的造血细胞和分离的正常对应物之间的关键相似性或差异。基于我们目前使用Notch配体Delta 1的人CD 34+干/祖细胞的产生和临床应用，我们还将测试良好表征的体外产生的干/祖细胞群体的体内生物学功能，并评估与“正常”分子谱的潜在偏离的意义（项目3）。为了进一步了解Notch和Wnt诱导的自我更新和分化影响的机制，我们将在ES/IPS和脐带血来源的CD 34+细胞的扩增群体中进行基因组规模的“多重”RNAi筛选（项目4）。该团队代表了Fred哈钦森癌症研究中心（FHCRC）和华盛顿大学（UW）在造血干细胞生物学，Wnt和Notch信号传导，表观遗传学，发育中基因表达的演变，基因组规模的RNAi技术以及ES和IPS方面的必要经验。我们期望与评估心脏和肺以及造血祖细胞的干细胞联盟的其他成员合作，提供专门知识以及用于调节Notch和Wnt信号传导的独特试剂。