Role and Regulation of a Novel, Developmentally Restricted Hematopoietic Stem Cell

新型发育受限造血干细胞的作用和调节

• 批准号: 10224171
• 负责人: CAMILLA FORSBERG
• 金额: $ 45.53万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10224171
• 关键词: Activities of Daily Living; Adult; Award; Biological Assay; Biological Models; Blood; CRISPR interference; Cell Lineage; Cells; Chromatin; Clonal Hematopoietic Stem Cell; Development; Disease; Epigenetic Process; Exposure to; Fetal Development; Fetal Liver; Gene Expression Profile; Genes; Genetic Transcription; Goals; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell subsets; Hematopoietic stem cells; Immune; Immune System Diseases; In Situ; Informal Social Control; Inherited; Life; Link; Longevity; Lymphoid; Lymphoid Cell; Malignant Neoplasms; Mediating; Molecular; Molecular Profiling; Output; Population; Prevention; Property; Regulation; Research; Role; Stress; System; Testing; Transgenic Model; Transplantation; Work; base; cancer therapy; cell type; epigenetic memory; experimental study; fetal; fetus cell; hematopoietic stem cell differentiation; hematopoietic stem cell fate; hematopoietic stem cell self-renewal; improved; in vivo; insight; molecular dynamics; mouse model; multipotent cell; novel; novel strategies; post-transplant; preservation; reconstitution; self-renewal; single-cell RNA sequencing; stem cell biology; stem cell fate; stem cell function; stem cell population; stem cells; tool; transcriptome

PROJECT SUMMARY/ABSTRACT Our long-term research goals are to understand the mechanisms that regulate stem cell fate decisions. In this first renewal application of this award, we propose to pursue the regulation of self-renewal and lineage potential of the developmentally restricted hematopoietic stem cells (drHSCs) that we discovered in the current award period. Although this population fulfills the most stringent criteria for functional HSC, their life-span during normal development is restricted to a limited developmental window. A functional HSC that does not persist into adulthood had never been observed before and therefore defines a novel wave of definitive hematopoiesis with a distinct endpoint. Here, we focus on understanding the contradictory regulation of drHSC self-renewal and multipotency: upon transplantation, drHSC self-renewal is induced, whereas their intrinsic lineage bias is preserved. Amazingly, the latter – lymphoid bias and exceptional B1a reconstitution potential – is maintained over many months even upon the repeated stress of serial transplantation. In contrast, a single, short-term exposure to stress induces the ability of drHSCs to persist long-term. We propose to pursue the epigenetic mechanisms governing this paradox. We will perform comprehensive molecular and cellular comparisons of drHSCs, co-existing fetal liver HSCs, and adult HSCs, and pursue rigorous functional analysis in competitive reconstitution assays. Importantly, we will couple single-cell transcriptional profiles with functional HSC capacity in efficient yet rigorous in vivo assays. Using CRISPRi/a-mediated transcriptional manipulation, we will directly test the requirements for reprogramming HSC self-renewal and lineage potential in vivo. Our transgenic models are uniquely suited for understanding how the core properties of HSCs – self-renewal and multilineage potential - are established during development and maintained for life and we are excited to put these powerful tools to work to pioneer developmental hematopoietic fate decisions.

项目摘要/摘要 我们的长期研究目标是了解调节干细胞脂肪决策的机制。在这个 我们提出的首次续签适用该奖项，我们建议追求自我更新和血统潜力的监管 我们在当前奖励中发现的开发受限制的造血干细胞（DRHSC） 期间。尽管该人口达到了功能性HSC的最严格标准，但在正常情况下它们的寿命 开发仅限于有限的发展窗口。一个不持续到的功能性HSC 以前从未观察过成年，因此定义 独特的端点。在这里，我们专注于理解DRHSC自我更新和 多能力：移植后，诱导了DRHSC自我更新，而它们的内在谱系偏见是 保留。令人惊讶的是，后者 - 淋巴偏见和杰出的B1A重建潜力 - 即使在连续移植的重复应力下，也有多个月的时间。相反，一个短期 暴露于压力会导致DRHSC长期持续存在的能力。我们建议追求表观遗传学 控制这个悖论的机制。我们将进行全面的分子和细胞比较 DRHSC，共存的胎儿肝HSC和成人HSC，以及竞争性的纯粹功能分析 重组测定法。重要的是，我们将将单细胞转录曲线与功能性HSC容量相结合 在有效但严格的体内测定中。使用CRISPRI/A介导的转录操作，我们将直接 测试在体内重编程HSC自我更新和谱系潜力的要求。我们的转基因模型 独特地适合理解HSC的核心特性（自我更新和多节势）如何 在开发过程中建立并维持生命，我们很高兴能将这些强大的工具放在 努力开拓发展性造血命运决定。