Mapping human hematopoietic stem cell development

绘制人类造血干细胞发育图谱

• 批准号: 10633115
• 负责人: Hanna Katri Annikki Mikkola
• 金额: $ 30.11万
• 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 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10633115
• 关键词: Address; Adult; Anatomy; 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; 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; Tissues; Transplantation; Umbilical Cord Blood; Validation; Yolk Sac; blood treatment; design; endothelial stem cell; 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; model organism; molecular marker; progenitor; programs; self-renewal; single-cell RNA sequencing; stem cell function; stem cells; stemness; timeline; 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）是推导HLA匹配或患者特异性造血茎的理想来源 细胞（HSC）治疗血液疾病。但是，产生自我更新的所有努力 来自PSC的造血干细胞（HSC）由于我们对机制的有限理解而失败了 在发展人类HSC中管理“茎”。尽管驱动血液规范的转录因子 相对理解，我们缺乏对定义HSC自我更新的程序的了解，以及为什么这些程序 PSC衍生的造血细胞中的程序失败。这，我们的目的是创建一个单元的转录组图 人体HSC在体内和ESC衍生的细胞体外产生的细胞。比较人类HSC的能力 指定HSC（AGM，可能是Pleceta和蛋黄）和蛋黄囊）和 在概念（胎儿肝脏和胎儿骨髓）中扩展并成熟到功能齐全的HSC中，将使我们能够 确定对HSC自我更新至关重要的途径，并区分人类HSC在体内发展 那些在体外发展的人。该分析将查明关键缺陷，这是ESC衍生功能不佳的基础 造血细胞，并为克服这些分子屏障提供新的溶液。由于缺乏HSC 可以可靠地预测人类HSC功能的表面标记，以帮助监测正确的分化 指定的HSC，我们为关键HSC调节因子HOXA5，MLLT5和HLF创建了HESC报告线 其表达在自我更新的人类HSC中高度丰富。然后，我们将比较体外得出的 候选HSC到人类造血组织的单细胞血管血管谱图随着HSC的出现 并在体内扩展。这种Shine-II RO1旨在解决体外HSC生成的关键障碍步骤。 这些研究可以扩展到机械研究，以了解人类HSC调节，并最终 转化研究以改善HSC移植。