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; 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.

项目摘要/摘要 造血干细胞（HSC）位于血液等级的顶部，能够产生所有血液 生物体的细胞。因此，它们的使用具有巨大的治疗潜力 体外疾病和HSC的产生是再生生物学的核心目的。尽管有这种临床需求，我们 缺乏使我们能够在体外产生HSC的协议，这些HSC可以长期植入和 多线输出。一个主要的障碍是我们对在体内如何制作HSC的不完全理解。为了 例子，尽管已经确定血细胞是从整个整个部位的多个部位的内皮细胞发展出来的 胚胎，我们仍然不知道哪些胚胎站点会产生长期HSC，也不知道原产地点如何影响 终身的干细胞功能或行为。此外，我们对内在和的理解有限 驱动血肿内皮细胞中功能异质性的外在提示。这个项目的建议要使用 强大的下一代条形码技术，以丰富我们对HSC和HSC胚胎起源的理解 血肿的内皮细胞状态会引起血液，使我们能够在体外利用这一过程。 鲍林博士在发育生物学和博士后培训中从事她的研究生工作 专注于开发用于执行单细胞，诱导细胞的下一代DNA条形码工具 体内谱系跟踪。 Bowling博士配备了此技能，计划使用创新的蜂窝条形码 技术首次对长寿祖细胞的精确内皮起源进行分类 哺乳动物胚胎（目标1）。此外，她将进行异质的深入表征 内皮细胞在胚胎中产生不同血细胞的内皮细胞状态（AIM 2）。从 这些实验有可能在发展领域回答重大的长期存在的问题 造血并改变我们对导致血液产生的步骤的基本理解，因此 彻底改变了HSC在体外生成的方案。 鲍林博士得到了一组导师和顾问小组的支持，这些研究员是世界一流的研究人员 血液学，发育生物学和技术发展。她的导师博士。费尔南多·卡马戈（Fernando Camargo）和 伦纳德·Zon（Leonard Zon Mentalship的记录。鲍林博士将获得她的进一步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，她将受益于携带 在波士顿儿童的科学刺激和资源丰富的环境中，她的研究计划 医院和哈佛医学院。该提案的目的将使保龄球博士能够以她的技能为基础 在测序数据集的计算分析中获得专家知识和诱导多能的使用 干细胞，准备过渡到独立性。结果，她将建立一个独特的利基市场 作为独立研究人员，解决造血发展中重要的临床问题。