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: 8107525
• 负责人: IRWIN D BERNSTEIN
• 金额: $ 120.96万
• 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/8107525
• 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）： 用于治疗目的的所需细胞类型的产生正在成为现实，它通过开发从胚胎干细胞（ESC），诱导的多能干细胞（IPSC）以及从孤立的成人干/祖细胞或分化细胞中直接“重新编程”到素质的干型干/祖细胞中的方法而成为现实。但是，实现这一目标将需要用于避免诱导永久遗传改变的治疗用数量的细胞，并确保衍生谱系合作的茎/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以直接分化和扩展反映正常发育状态的细胞类型的种群。作为这种方法的模型，我们专注于缺口和WNT途径以及特征良好的造血系统，以产生造血干细胞（HSC）。通过扩大脐带血的茎/祖细胞以及我们在临床环境中成功应用这些细胞，可以证明扩大治疗有用的茎/祖细胞扩展的可行性。具体而言，我们将研究Notch和Wnt信号传导在胚胎中产生第一个HSC的要求和时机，以指导我们在体内产生这些细胞的努力。我们将通过增强对确定造血的茎/祖细胞的血液生成内皮前体的分化以及促进这些多能茎/祖细胞种群的自我生产（项目1）来产生ES和IPS来源的HSC。为了评估人类ES和IPS衍生的茎/祖细胞的治疗有用性，我们将确定它们保留转录，染色质和DNA甲基化以及功能性景观（项目2）。这些研究将与有关Wnt和Notch在扩展和正确分化心脏干细胞的作用的协作链接应用中所描述的研究。