Identification of novel regulators of HSC specification and maturation

HSC 规范和成熟的新型调节剂的鉴定

• 批准号: 10665070
• 负责人: Teresa V Bowman
• 金额: $ 29.21万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-23 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665070
• 关键词: Biological Assay; Blood; CRISPR/Cas technology; Candidate Disease Gene; Cell Separation; Cells; Clinical; Competence; Coupled; Data; Dedications; Development; Developmental Biology; Dissection; Embryo; Embryonic Development; Endothelial Cells; Environment; Gene Expression; Generations; Genes; Genetic; Goals; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic Cell Production; Hematopoietic Stem Cell Specification; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; Immunologics; In Situ; In Vitro; Kinetics; Knowledge; Laboratories; Larva; Measures; Mediating; Modeling; Mus; Mutagenesis; Natural regeneration; Pathway interactions; Patients; Positioning Attribute; Process; Production; Property; Public Health; RNA Processing; Regulation; Replacement Therapy; Research; Specific qualifier value; Stem Cell Development; System; Testing; Time; Transplantation; Zebrafish; experience; hematopoietic stem cell formation; hematopoietic stem cell self-renewal; hematopoietic transplantation; hemogenic endothelium; improved; in vivo; induced pluripotent stem cell; novel; novel strategies; progenitor; programs; regenerative; self-renewal; single-cell RNA sequencing; stem cells; zebrafish development

Identification of novel regulators of HSC specification and maturation Project Abstract Hematopoietic stem cells (HSCs) are defined by self-renewal and multilineage differentiation capacities. HSCs sustain lifelong hematopoiesis and are exploited for hematopoietic transplantation to replace damaged or diseased blood systems in patients with hematologic disorders. Despite the widespread usage, transplantation remains unattainable for many due to the paucity of appropriate immunologically matched donors. De novo production of HSCs from patient-specific induced pluripotent stem cells (iPSCs) has been proposed as a means to circumvent this problem, but it remains an extremely inefficient process. Current approaches result in the abundant creation of hematopoietic progenitors that possess limited differentiation abilities and lack sufficient transplantation capacity. During development, there is simultaneous de novo production of these limited-capacity progenitors alongside HSCs. One of the major limitations in the field is the inability to distinguish these cells at the time of formation, which makes it difficult to define the factors distinctly critical for HSC generation. Through single-cell RNA-sequencing of nascent hematopoietic stem and progenitor cells from developing zebrafish, we identified genes that discriminate HSCs from progenitors at the time of their formation. We also developed new in situ lineage-tracing and regenerative assays in zebrafish larvae to define functional differences between the differentiation kinetics and self-renewal of these cell subsets. Here, we plan to exploit the genetic malleability of zebrafish coupled with our novel assays to discover new regulators uniquely required for HSC formation. The SHINE-II mechanism (PAS-18-730) is dedicated to supporting new research directions in their early stages. Consistent with this aim, this project takes our laboratory in a new direction away from our focus on RNA processing in hematopoiesis and into the realm of understanding de novo production of bona fide HSCs. Our experience in zebrafish genetics and developmental biology along with our establishment of new assays to functionally distinguish nascent HSCs from HSC-independent progenitors make us well-positioned to accomplish our goal of identifying novel factors that uniquely regulate HSC specification and functional maturation. Identifying this program will provide fundamental knowledge on the acquisition of self-renewal and multipotency attributes and could provide guidance to improve ex vivo HSC production from mammalian iPSCs.

HSC特化和成熟的新调控因子的鉴定 项目摘要 造血干细胞（HSCs）具有自我更新和多向分化的能力。HSCs 维持终生造血，并用于造血移植以替代受损或 血液系统疾病患者的血液系统。尽管移植被广泛使用， 由于缺乏适当的免疫匹配的供体，许多人仍然无法获得。从头 已经提出从患者特异性诱导多能干细胞（iPSC）产生HSC作为一种手段 来规避这个问题，但这仍然是一个极其低效的过程。目前的方法导致 造血祖细胞的大量产生，其具有有限的分化能力并且缺乏足够的 移植能力。在开发过程中，同时从头生产这些有限容量的 与造血干细胞一起。该领域的主要限制之一是无法区分这些细胞， 形成的时间，这使得很难定义对HSC生成明显关键的因素。通过 对来自发育中斑马鱼的新生造血干细胞和祖细胞进行单细胞RNA测序，我们 确定了在造血干细胞形成时将其与祖细胞区分开来的基因。我们还开发了新的 在斑马鱼幼鱼中进行原位谱系追踪和再生试验，以确定 这些细胞亚群的分化动力学和自我更新。在这里，我们计划利用基因的可塑性， 斑马鱼结合我们的新的测定，以发现HSC形成独特需要的新的调节剂。的 SHINE-II机制（PAS-18-730）致力于在早期阶段支持新的研究方向。 与此目标一致，该项目将我们的实验室带到一个新的方向，远离我们对RNA的关注 在造血过程中的处理，并进入理解真正的造血干细胞的从头生产的领域。我们 沿着我们建立了新的检测方法， 在功能上区分新生的HSC和HSC独立的祖细胞使我们能够很好地完成 我们的目标是确定新的因素，独特的调节HSC的特化和功能成熟。 确定这一计划将提供基本知识的收购自我更新和多能性 这些特性可以为改善哺乳动物iPSC的离体HSC生产提供指导。