Expansion of Cardiac and Hematopoietic Pregenitors by Wnt and Notch

Wnt 和 Notch 扩增心脏和造血祖细胞

• 批准号: 7939768
• 负责人: IRWIN D BERNSTEIN
• 金额: $ 119.73万
• 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: 2009-09-30 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7939768
• 关键词: Address; Adult; Affect; Behavioral; Biological Preservation; Cardiac; Cell Count; Cells; Chromatin; Clinical; Clinical Trials; Commit; DNA Methylation; Derivation procedure; Development; Embryo; Endothelium; Engineering; Ensure; Epigenetic Process; Generations; Genes; Goals; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Human; Link; Malignant - descriptor; Methods; Modeling; Moon; Mus; Mutation; Notch and Wnt Signaling Pathway; Pathway interactions; Pear; Physiological; Population; Role; Signal Transduction; Staging; Stem cells; Stimulus; Testing; Therapeutic; Therapeutic Uses; Therapy Clinical Trials; Time; Umbilical Cord Blood; cell type; embryonic stem cell; human embryonic stem cell; in vivo; induced pluripotent stem cell; method development; mutant; notch protein; progenitor; self-renewal; stem; stem cell fate specification; stem cell population

DESCRIPTION (provided by applicant): The generation of desired cell types for therapeutic purposes is becoming a reality with the development of methods for deriving such cells from embryonic stem cells (ESC), induced pluripotent stem cells (iPSC), as well as from isolated adult stem/progenitor cells or differentiated cells that are directly "reprogrammed" into lineage-specific stem/progenitor cells. Realizing this goal, however, will require methods for deriving therapeutically useful numbers of cells that avoid inducing permanent genetic alterations, and ensure the behavioral fidelity of derived lineage-committed stem/progenitor cells. To address this issue, we propose to test our hypotheses that pathways regulating normal development can be manipulated to direct differentiation and expansion of populations of cell types that reflect normal developmental states. As a model for this approach, we focus on the Notch and Wnt pathways and the well-characterized hematopoietic system to generate hematopoietic stem cells (HSC). The feasibility of expanding therapeutically useful stem/progenitor cells is demonstrated by our expansion of cord blood-derived stem/progenitor cells and by our successful application of these cells in a clinical setting. Specifically, we will examine the requirement for and timing of Notch and Wnt signaling in generating the first HSCs in the embryo, to guide our efforts to produce these cells ex vivo. We will generate ES- and iPS-derived HSC by enhancing differentiation towards hemogenic endothelial precursors of definitive hematopoietic stem/progenitor cells, and by promoting selfrenewal of these multipotent stem/progenitor populations (Project 1). To assess the therapeutic usefulness of human ES- and iPS-derived stem/progenitor cells, we will determine their preservation of the transcriptional, chromatin and DNA methylation and functional landscapes (Project 2). These studies will interface with those described in the collaborative linked application on the role of Wnt and Notch in expansion and proper differentiation of cardiac stem cells.

描述（由申请人提供）： 随着从胚胎干细胞（ESC）、诱导多能干细胞（iPSC）以及从分离的成体干/祖细胞或直接“重编程”为谱系特异性干/祖细胞的分化细胞中衍生出此类细胞的方法的开发，用于治疗目的的所需细胞类型的产生正在成为现实。然而，实现这一目标将需要获得治疗上有用的细胞数量的方法，以避免诱导永久性遗传改变，并确保衍生的谱系定向干/祖细胞的行为保真度。为了解决这个问题，我们建议测试我们的假设，即可以操纵调节正常发育的途径来指导反映正常发育状态的细胞类型群体的分化和扩增。作为这种方法的模型，我们重点关注 Notch 和 Wnt 通路以及特征明确的造血系统，以生成造血干细胞 (HSC)。我们对脐带血干细胞/祖细胞的扩增以及这些细胞在临床环境中的成功应用证明了扩增具有治疗用途的干/祖细胞的可行性。具体来说，我们将检查在胚胎中生成第一个 HSC 时 Notch 和 Wnt 信号传导的要求和时间，以指导我们在体外生产这些细胞的努力。我们将通过增强定向造血干/祖细胞向造血内皮前体的分化，并促进这些多能干/祖细胞群的自我更新，产生 ES 和 iPS 衍生的 HSC（项目 1）。为了评估人类 ES 和 iPS 衍生的干/祖细胞的治疗用途，我们将确定它们对转录、染色质和 DNA 甲基化以及功能景观的保存（项目 2）。这些研究将与协作链接申请中描述的关于 Wnt 和 Notch 在心脏干细胞扩增和正确分化中的作用的研究相结合。