Improving the interaction of hematopoietic stem cells with the perivascular niche to promote engraftment

改善造血干细胞与血管周围微环境的相互作用以促进植入

• 批准号: 10930184
• 负责人: Owen James Tamplin
• 金额: $ 53.76万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-20 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10930184
• 关键词: ATAC-seq; Address; Adult; Aorta; Area; Behavior; Binding; Biological Models; Blood Vessels; Bone Marrow; Bypass; Cell Adhesion; Cell Adhesion Molecules; Cell Communication; Cell Differentiation process; Cell Maturation; Cell Ontogeny; Cell Therapy; Cells; Chromatin; Circulation; Clinical; Coculture Techniques; Complex; Data; Derivation procedure; Development; Dorsal; Electron Microscopy; Embryo; Endothelial Cells; Engraftment; Erythro; Event; Extracellular Matrix; Fetal Liver; Genetic; Goals; Growth; Hematological Disease; Hematology; Hematopoiesis; Hematopoietic Neoplasms; Hematopoietic stem cells; Human; Image; In Vitro; Integrin alpha Chains; Integrins; Investigation; Kidney; Knowledge; Mammals; Marrow; Mediating; Methods; Modeling; Mus; Myelogenous; Non-Malignant; PIK3CG gene; Pathway interactions; Patient-Focused Outcomes; Patients; Phenotype; Population; Positioning Attribute; Procedures; Process; Protocols documentation; Research; Research Proposals; Role; SU 5416; Signal Transduction; Signaling Molecule; Specific qualifier value; Stem cell transplant; Testing; Therapeutic; Tissues; Translating; Transplantation; Travel; Up-Regulation; Vascular Endothelial Growth Factor C; Vascular Endothelial Growth Factors; Vertebrates; Zebrafish; cell behavior; cell motility; cell type; curative treatments; fetal; genetic signature; hematopoietic differentiation; hematopoietic tissue; hematopoietic transplantation; hemogenic endothelium; improved; improved outcome; in vivo; inhibitor; innovation; knock-down; light microscopy; live cell imaging; loss of function; lymphatic development; lymphatic vessel; migration; mouse model; mutant; novel; progenitor; programs; receptor; response; single-cell RNA sequencing; small molecule; stem cells; tool

PROJECT SUMMARY/ABSTRACT Hematopoietic stem and progenitor cell (HSPC) transplantation is a curative treatment for many blood diseases and cancers. However, these procedures still need to be optimized to improve patient outcomes and survival. HSPCs reside in a microenvironment surrounded by niche cells that help regulate their function. Our research proposal seeks to address fundamental questions regarding the cellular interactions between HSPCs and niche cells. During development in the embryo, HSPCs move through different tissues and have changing requirements for contact with the microenvironment. HSPCs first arise in the dorsal aorta, a large vessel in the embryo, and are then released into circulation. Next, HSPCs migrate to the fetal liver where the population of cells expands exponentially via symmetric divisions. Finally, HSPCs migrate again to colonize the bone marrow where they will remain throughout adulthood. As HSPCs are migrating between these different hematopoietic tissues, they are also becoming more mature and are programmed towards their adult quiescent state. We use zebrafish and mice as model systems that are highly conserved with humans and have many genetic tools for functional testing and live imaging of cellular behaviors. In zebrafish, we have found a novel role for vascular endothelial growth factor c (vegfc) in the release of HSPCs from the dorsal aorta. Vegfc also regulates a fate decision in pre-HSPCs that determines if they will become a stem cell or a different type of progenitor cell. At later stages, we found a programming function for integrin alpha 4 (itga4) in the caudal hematopoietic tissue (CHT), the zebrafish equivalent of the fetal liver. We hypothesize that the timing of HSPC contact and transitions between developmental niches is critical for their correct programming. We will address this hypothesis through the following Specific Aims: 1) Define the role of vegfc in HSPC transition and release from the dorsal aorta. 2) Determine how itga4-dependent contact with the CHT niche programs HSPCs as they transition from fetal-like to adult stages. We will perform in vivo genetic knockdown and small molecule treatments, together with live imaging, to understand the dynamic interactions between HSPCs and niche cells. We will then translate these results to an in vitro vascular niche that is used to differentiate cells into transplantable HSPCs that could lead to cell-based therapies for patients.

项目总结/摘要 造血干/祖细胞移植是治疗多种血液病的有效方法 和癌症。然而，这些程序仍然需要优化，以改善患者的结果和生存。 HSPCs位于微环境中，周围是帮助调节其功能的小生境细胞。我们的研究 一项提案旨在解决有关HSPC和小生境之间细胞相互作用的基本问题， 细胞在胚胎发育过程中，HSPCs在不同组织中移动， 与微环境接触的要求。HSPCs首先出现在背主动脉中，背主动脉是心脏中的大血管。 胚胎，然后释放到循环中。接下来，HSPC迁移到胎儿肝脏，在那里HSPC的群体被转移到胎儿肝脏。 细胞通过对称分裂以指数方式扩张。最后，HSPCs再次迁移到骨髓中 在那里他们将度过整个成年期。当HSPC在这些不同的造血细胞之间迁移时， 组织，它们也变得更加成熟，并被编程为它们的成年静止状态。我们使用 斑马鱼和小鼠作为模型系统，与人类高度保守，并有许多遗传工具， 功能测试和细胞行为的实时成像。在斑马鱼中，我们发现了一个新的作用， 内皮生长因子c（vegfc）在HSPC从背主动脉释放中的作用。Vegfc还调节了一种命运， 在前HSPC中，决定它们是否将成为干细胞或不同类型的祖细胞的决定。在 后期，我们发现尾部造血组织中整合素α 4（ITGA 4）的编程功能 (CHT)斑马鱼相当于胎儿肝脏。我们假设HSPC接触和转换的时间 对他们的正确规划至关重要。我们将通过以下方式来解决这一假设： 以下具体目的：1）确定vegfc在HSPC转化和从背侧释放中的作用 主动脉2)确定ITGA 4依赖性接触如何与CHT利基程序HSPC，因为他们的过渡 从胎儿到成人阶段。我们将进行体内基因敲除和小分子治疗， 结合活体成像，了解HSPCs和小生境细胞之间的动态相互作用。然后我们将 将这些结果转化为用于将细胞分化为可移植HSPC的体外血管生态位 这将为患者带来基于细胞的治疗。