Mapping human hematopoietic stem cell development

绘制人类造血干细胞发育图谱

• 批准号: 10435434
• 负责人: Hanna Katri Annikki Mikkola
• 金额: $ 30.42万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10435434
• 关键词: Address; Adult; Anatomy; Animal Model; Blood; Blood Vessels; Bone Marrow; CD34 gene; Cell Lineage; Cell Maturation; Cell Ontogeny; Cell surface; Cells; Code; Communities; Conceptus; Critical Pathways; Data; Defect; Development; EPS15 gene; Embryo; Endothelium; Engraftment; Fetal Liver; First Pregnancy Trimester; Generations; Genes; Goals; Growth; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Specification; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; Human Biology; Human Development; In Vitro; Knowledge; Lead; Liver; Location; MLLT3 gene; Maps; Medial; Molecular; Molecular Profiling; Monitor; Mus; Organ; Patients; Placenta; Pluripotent Stem Cells; Process; Protocols documentation; Regenerative Medicine; Regulation; Reporter; Resolution; Resources; Safety; Second Pregnancy Trimester; Source; Specific qualifier value; Stem Cell Development; Surface; Time; TimeLine; Tissues; Transplantation; Umbilical Cord Blood; Validation; Yolk Sac; design; fetal; hematopoietic stem cell fate; hematopoietic stem cell self-renewal; hematopoietic tissue; hemogenic endothelium; human embryonic stem cell; human fetus tissue; human pluripotent stem cell; improved; in vivo; induced pluripotent stem cell; migration; molecular marker; progenitor; programs; self-renewal; single-cell RNA sequencing; stem cell function; stem cell therapy; stemness; tissue stem cells; transcription factor; transcriptome; translational study

SUMMARY Pluripotent stem cells (PSC) are an ideal source for deriving HLA-matched or patient specific hematopoietic stem cells (HSC) for the treatment of blood disorders. However, all the efforts in producing self-renewing hematopoietic stem cells (HSC) from PSC have failed due to our limited understanding of the mechanisms that govern “stemness” in developing human HSC. Although the transcription factors that drive blood specification are relatively well understood, we lack knowledge of the programs that define HSC self-renewal, and why these programs fail in PSC derived hematopoietic cells. Thus, we aim to create a single cell transcriptome map of human HSC ontogeny in vivo and ESC derived cells generated in vitro. The ability to compare human HSC from the critical stages of ontogeny when HSCs are specified (AGM, and possibly the placenta and yolk sac) and expand and mature into fully functional HSC in the conceptus (fetal liver and fetal bone marrow) will allow us to identify pathways that are critical for HSC self-renewal, and that distinguish human HSCs developing in vivo from those that develop in vitro. This analysis will pinpoint key defects that underlie the poor function of ESC derived hematopoietic cells, and offer new solutions for overcoming these molecular barriers. As there is a lack of HSC surface markers that would reliably predict human HSC function, to help monitor the differentiation of properly specified HSCs, we created hESC reporter lines for critical HSC regulatory factors HOXA5, MLLT5 and HLF whose expression is highly enriched in self-renewing human HSCs. We will then compare the in vitro derived candidate HSC to the single cell hemato-vascular lineage map of human hematopoietic tissues as HSCs emerge and expand in vivo. This SHINE-II RO1 is designed to address critical hurdle steps in in vitro HSC generation. These studies can be expanded to mechanistic studies to understand human HSC regulation, and ultimately to translational studies to improve HSC transplantation.

概括 多能干细胞 (PSC) 是获得 HLA 匹配或患者特异性造血干细胞的理想来源 细胞（HSC）用于治疗血液疾病。然而，所有的努力都是为了产生自我更新 由于我们对造血干细胞 (HSC) 的机制了解有限，因此从 PSC 中提取造血干细胞 (HSC) 失败了。 控制人类 HSC 发育的“干性”。尽管驱动血液规格的转录因子 相对来说，我们对 HSC 自我更新的定义程序缺乏了解，以及为什么这些程序 PSC 衍生的造血细胞中的程序失败。因此，我们的目标是创建单细胞转录组图谱 体内人类 HSC 个体发育和体外产生 ESC 衍生细胞。比较人类 HSC 的能力 当 HSC 被指定时，个体发育的关键阶段（AGM，可能还有胎盘和卵黄囊）和 在概念（胎儿肝脏和胎儿骨髓）中扩展并成熟为功能齐全的 HSC 将使我们能够 确定对 HSC 自我更新至关重要的途径，并将人类 HSC 与体内发育的 HSC 区分开来 那些在体外发育的。该分析将查明导致 ESC 衍生功能不佳的关键缺陷 造血细胞，并为克服这些分子障碍提供新的解决方案。由于缺乏HSC 能够可靠地预测人类 HSC 功能的表面标记，以帮助监测正确的分化 针对特定的 HSC，我们针对关键 HSC 调节因子 HOXA5、MLLT5 和 HLF 创建了 hESC 报告基因系 其表达在自我更新的人类 HSC 中高度富集。然后我们将比较体外衍生的 随着 HSC 的出现，候选 HSC 进入人类造血组织的单细胞血管谱系图谱 并在体内扩展。 SHINE-II RO1 旨在解决体外 HSC 生成中的关键障碍步骤。 这些研究可以扩展到机制研究，以了解人类 HSC 的调节，并最终 改善 HSC 移植的转化研究。