Cellular barcoding of developmental hematopoiesis

发育造血的细胞条形码

• 批准号: 10669239
• 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 2024-02-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10669239
• 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; Location; Maps; Mentors; Mentorship; Molecular; Molecular Profiling; Mus; Names; Natural regeneration; Organism; Outcome; Output; Pediatric Hospitals; Phase; Physiological; Pluripotent Stem Cells; Postdoctoral Fellow; Preparation; Process; Production; Productivity; Property; Protocols documentation; Publishing; Records; Research; Research Personnel; Resolution; Resources; Role; Route; Site; System; Techniques; Technology; Testing; Therapeutic; Time; Training; Work; 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.

项目总结/摘要 造血干细胞（HSC）位于血液层级的顶端，能够产生所有血液， 一个有机体的细胞。因此，它们的使用对于血液处理具有巨大的治疗潜力。 疾病，并且在体外产生HSC是再生生物学的中心目标。尽管有这种临床需求，我们 缺乏允许我们在体外有效产生能够长期植入的HSC的方案， 多谱系输出。一个主要的障碍是我们对HSC如何在体内产生的不完全理解。为 例如，尽管已经确定血细胞在整个血管的多个部位从内皮细胞发育而来， 尽管我们在胚胎中发现了HSC，但我们仍然不知道哪些胚胎部位产生长期的HSC，也不知道起源部位如何影响HSC。 干细胞的功能或行为。此外，我们对内在和内在的理解是有限的， 驱动生血内皮细胞功能异质性的外在因素。该项目建议使用 强大的下一代条形码技术，以丰富我们对HSC胚胎起源的理解， 生血内皮细胞产生血液的状态，使我们能够在体外利用这个过程。 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，用于进行单细胞，诱导细胞， 体内谱系追踪。有了这种技能，Bowling博士计划使用创新的细胞条形码技术， 技术，第一次，目录中的长寿血液祖细胞的精确内皮起源， 哺乳动物胚胎（Aim 1）。此外，她还将深入描述 在胚胎中产生不同血细胞的内皮细胞状态（目的2）。知识产生于 这些实验有可能回答发展领域长期存在的主要问题。 造血和改变我们的基本理解的步骤，导致血液生成，因此， 彻底改变体外HSC生成的方案。 Bowling博士得到了一个由导师和顾问组成的小组的支持，他们是世界级的研究人员， 血液学、发育生物学和技术开发。她的导师费尔南多·卡马戈博士和 伦纳德宗在造血领域做出了卓越的贡献，也有杰出的田径运动- 记录导师。鲍灵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、Trista North和Berthold Gottgens博士。最后，她将受益于携带 在波士顿儿童医院的科学刺激和资源丰富的环境中， 医院和哈佛医学院。本提案的目的将使鲍林博士能够利用她的技能， 获得测序数据集的计算分析和诱导多能基因的使用方面的专业知识 干细胞，为她过渡到独立做准备。因此，她将建立一个独特的利基， 作为一名独立的研究人员，解决造血发育中重要的临床相关问题。