Cellular barcoding of developmental hematopoiesis

发育造血的细胞条形码

• 批准号: 10506135
• 负责人: Sarah Bowling
• 金额: $ 15.38万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10506135
• 关键词: Adult; Anatomy; Aorta; Area; Automobile Driving; Bar Codes; Behavior; Blood; Blood Cells; Blood flow; Bone Marrow Purging; Boston; CRISPR/Cas technology; Catalogs; Cell Lineage; Cells; Clinic; Clinical; Computer Analysis; Cues; DNA; Data; Data Set; Derivation procedure; Development; Developmental Biology; Embryo; Endothelial Cells; Endothelium; Engraftment; Environment; Epigenetic Process; Fetal Liver; Generations; Genes; Genetic Transcription; Goals; Growth; Hematological Disease; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Heterogeneity; In Vitro; Investigation; Knowledge; Life; Light; Location; Maps; Mentors; Mentorship; Molecular; Molecular Profiling; Mus; Names; Natural regeneration; Organism; Outcome; Output; Pediatric Hospitals; Phase; Physiological; Pluripotent Stem Cells; Preparation; Process; Property; Protocols documentation; Publishing; Records; Research; Research Personnel; Resolution; Resources; Role; Route; Site; System; Techniques; Technology; Testing; Therapeutic; Time; Training; Work; base; blood formation; blood treatment; career development; cell behavior; chemotherapy; clinically relevant; driving force; endothelial stem cell; experimental study; hematopoietic stem cell emergence; hematopoietic stem cell formation; hemogenic endothelium; human pluripotent stem cell; improved; in vivo; induced pluripotent stem cell; innovation; leukemia/lymphoma; medical schools; mouse model; multiple omics; next generation; novel strategies; post-doctoral training; progenitor; programs; regenerative biology; skills; stem cell function; stem cells; technology development; tool

PROJECT SUMMARY/ ABSTRACT Hematopoietic stem cells (HSCs) lie at the top of the blood hierarchy and are capable of giving rise to all blood cells of an organism. Consequently, their use has enormous therapeutic potential for the treatment of blood diseases, and generation of HSCs in vitro is a central aim in regenerative biology. Despite this clinical need, we lack protocols that allow us to efficiently generate HSCs in vitro that are capable of long-term engraftment and multi-lineage output. A major hindrance is our incomplete understanding of how HSCs are made in vivo. For instance, although it is established that blood cells develop from endothelial cells in multiple sites throughout the embryo, we still do not know which embryonic sites produce long-term HSCs, nor how site of origin impacts on life-long stem cell function or behavior. Furthermore, we are limited in our understanding of the intrinsic and extrinsic cues driving functional heterogeneity in hemogenic endothelial cells. This project proposes to use powerful next-generation barcoding technology to enrich our understanding of the embryonic origin of HSCs and the hemogenic endothelial cell states that give rise to blood to allow us to harness the process in vitro. Dr. Bowling conducted her graduate work in developmental biology and during her postdoctoral training has focused on the development of next-generation DNA barcoding tools for performing single cell, inducible cell lineage tracing in vivo. Equipped with this skillset, Dr. Bowling plans to use innovative cellular barcoding techniques to, for the first time, catalog the precise endothelial origins of long-lived blood progenitors in the mammalian embryo (Aim 1). Furthermore, she will perform in-depth characterization of the heterogeneous endothelial cell states that give rise to distinct blood cells in the embryo (Aim 2). The knowledge generated from these experiments have the potential to answer major, long-standing questions in the field of developmental hematopoiesis and transform our basic understanding of the steps leading to blood generation, and therefore revolutionize protocols for HSC generation in vitro. Dr. Bowling is supported by a panel of mentors and consultants who are world-class researchers in hematology, developmental biology, and technology development. Her mentors Drs. Fernando Camargo and Leonard Zon have made exceptional contributions to the field of hematopoiesis and also have outstanding track- records for mentorship. Dr. Bowling will gain further scientific training and career development support from her scientific committee: Drs Jay Shendure, Trista North and Berthold Gottgens. Finally, she will benefit from carrying out her research program in the scientifically stimulating and resource-rich environment of Boston Children’s Hospital and Harvard Medical School. The aims in this proposal will allow Dr. Bowling to build on her skills to gain expert knowledge in the computational analysis of sequencing datasets and the use of induced pluripotent stem cells, in preparation for her transition to independence. As a result, she will establish a unique niche for resolving important, clinically-relevant questions in hematopoietic development as an independent researcher.

项目概要/摘要 造血干细胞 (HSC) 位于血液层次结构的顶端，能够产生所有血液 经过检查，它们的用途对于治疗血液具有巨大的治疗潜力。 尽管存在这种临床需求，但体外生成 HSC 是再生生物学的中心目标。 缺乏使我们能够在体外有效生成能够长期移植的 HSC 的方案 多谱系输出的一个主要障碍是我们对体内 HSC 的形成方式不完全了解。 例如，尽管已经确定血细胞是由整个体内多个部位的内皮细胞发育而来的 胚胎中，我们仍然不知道哪些胚胎位点产生长期 HSC，也不知道起源位点如何影响 此外，我们对干细胞的内在和行为的理解有限。 该项目建议使用驱动造血内皮细胞功能异质性的外在线索。 强大的下一代条形码技术可丰富我们对 HSC 胚胎起源的理解 产生血液的造血内皮细胞状态使我们能够在体外利用这一过程。 Bowling 博士在发育生物学和博士后培训期间进行了她的研究生工作 专注于开发下一代 DNA 条形码工具，用于执行单细胞、诱导细胞 配备了这套技能后，鲍林博士计划使用创新的细胞条形码。 首次对长寿血液祖细胞的精确内皮起源进行分类的技术 此外，她还将对异质性进行深入的表征。 内皮细胞状态在胚胎中产生不同的血细胞（目标 2）。 这些实验有可能回答发育领域长期存在的重大问题 造血作用并改变了我们对血液生成步骤的基本理解，因此 彻底改变 HSC 体外生成方案。 Bowling 博士得到了一组导师和顾问的支持，他们都是世界一流的研究人员 她的导师费尔南多·卡马戈博士和血液学、发育生物学和技术开发。 伦纳德·佐恩（Leonard Zon）在造血领域做出了非凡的贡献，也拥有出色的田径成绩—— 鲍林博士将从她那里获得进一步的科学培训和职业发展支持。 科学委员会：Jay Shendure 博士、Trista North 和 Berthold Gottgens 最后，她将从携带中受益。 在波士顿儿童医院科学刺激且资源丰富的环境中开展她的研究项目 该提案的目的是让鲍林博士能够利用她的技能来实现以下目标： 获得测序数据集计算分析和诱导多能的使用方面的专业知识 干细胞，为她过渡到独立做好准备，因此，她将为她建立一个独特的利基市场。 作为独立研究人员解决造血发育中重要的临床相关问题。