Identification of novel regulators of HSC specification and maturation

HSC特异性和成熟度新调节子的鉴定 项目摘要 造血干细胞(HSCs)具有自我更新和多向分化的能力。肝星状细胞 维持终身造血，并用于造血移植，以取代受损的或 血液病患者的血液系统病变。尽管广泛使用，但移植 由于缺乏适当的免疫匹配捐赠者，许多人仍然无法获得。从头开始 从患者特异性的诱导多能干细胞(IPSCs)中生产HSCs已被提出作为一种手段 以绕过这一问题，但这仍然是一个效率极低的过程。目前的方法导致了 造血祖细胞分化能力有限而缺乏足够的造血祖细胞 移植能力。在开发过程中，这些有限的产能同时从头开始生产。 造血干细胞旁的祖细胞。该领域的主要限制之一是无法区分这些细胞 形成时间，这使得很难明确地定义对HSC产生至关重要的因素。穿过 斑马鱼发育中的新生造血干和祖细胞的单细胞RNA测序 在HSCs形成时识别了区分它们与祖细胞的基因。我们还开发了新的 斑马鱼幼体的原位谱系追踪和再生分析以确定两者之间的功能差异 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延展性 斑马鱼与我们的新检测相结合，以发现形成HSC所独特需要的新的调节剂。这个 SIRE-II机制(PAS-18-730)致力于在早期阶段支持新的研究方向。 与这个目标一致，这个项目将我们的实验室带向一个新的方向，远离我们对rna的关注。 在造血过程中的过程，并进入理解真正的造血干细胞的从头产生的领域。我们的 在斑马鱼遗传学和发育生物学方面的经验以及我们建立的新的检测方法 在功能上区分新生的HSC和独立于HSC的祖细胞，使我们处于有利地位，能够实现 我们的目标是确定独特调节HSC规格和功能成熟的新因素。 确定该计划将提供有关获得自我更新和多功能性的基础知识 并可为提高哺乳动物IPSCs体外培养的HSC产量提供指导。

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.