Cellular and molecular analyses of hematopoietic stem cell [HSC] interactions with bone marrow niches to improve HSC engraftment for transplantation and tolerance induction

造血干细胞 [HSC] 与骨髓微环境相互作用的细胞和分子分析，以改善 HSC 植入移植和耐受诱导

• 批准号: 9753220
• 负责人: IRVING L. WEISSMAN
• 金额: $ 37.43万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9753220
• 关键词: Acute Myelocytic Leukemia; Antibodies; Autoimmune Process; Back; Binding; Biological Assay; Biology; Blood; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; CXCR4 gene; Cartilage; Cell Adhesion Molecules; Cell Communication; Cell Count; Cells; Cellular Structures; Characteristics; Clonal Hematopoietic Stem Cell; Data; Disease; Dysmyelopoietic Syndromes; Elderly; Emigrations; Endothelial Cells; Endothelium; Engineering; Engraftment; Environment; Frequencies; Gene Expression; Genes; Goals; Grant; Health; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic Stem Cell subsets; Hematopoietic stem cells; Home environment; Homing; Human; Imagery; Immigration; In Vitro; Insulin-Dependent Diabetes Mellitus; Integrins; Knock-out; Knowledge; Lead; Learning; Location; Lymphopoiesis; Malignant Neoplasms; Marrow; Methods; Molecular; Molecular Analysis; Mus; Mutate; Mutation; Myelogenous; Myelopoiesis; Myeloproliferative disease; Organ; Pathology; Phenotype; Physiological; Population; Proteome; Regimen; Regulation; Reporter; Research; Role; Running; Side; Signal Transduction; Specificity; Stem Cell Development; Stem cells; Stromal Cells; Supporting Cell; Surface; Systemic Lupus Erythematosus; T-Lymphocyte; Tissue Donors; Toxic effect; Transplantation; Transplantation Tolerance; Venous; allotransplant; bone; cell type; chemokine; clinical application; conditioning; cytokine; experimental study; extracellular; hematopoietic stem cell expansion; hematopoietic stem cell formation; hematopoietic stem cell niche; improved; improved outcome; insight; intercellular communication; leukemia; mutant; normal aging; peripheral blood; progenitor; reconstitution; skeletal; stem cell biology; young adult

Project Summary We have shown that purified hematopoietic stem cells (HSC), first isolated by us, can be allotransplanted without GvH (they lack T cells), block autoimmune type 1 diabetes (and Systemic Lupus Erythematosus), and induce transplant tolerance to HSC donor tissues/organs. However, purified HSC transplantation is not used, in part due to the toxicity of the conditioning regimens. Therefore, it is vital that we improve our understanding of how HSCs interact with their local supporting environment—the hematopoietic niche. The goal of this research is to further identify and characterize the cellular and molecular components of the bone marrow (BM) HSC niche, and to understand how the ongoing inter-cellular communication between HSCs and their niche regulates their homing, anchoring, survival and function in health and in blood diseases. In previous versions of this grant we established antibodies in place of toxic regimens to condition recipients and will further modify the regimens to improve engraftment and accelerate reconstitution. Further, understanding the HSC-niche interaction has implications for stem cell competitions we have elucidated in pathology, for example, in hematopoietic disorders such as myelodysplastic syndrome (MDS), myeloproliferative neoplasms (MPN), and expansion of mutated clones to give clonal hematopoiesis, some that give rise to acute myeloid leukemia (AML). In the run-up to leukemia and MDS, clones of HSCs with pre-cancer mutations outcompete normal HSCs, but the total number of HSCs appears not to change, implying control of HSC numbers by the niche. The natural recirculation of HSCs allows highly competitive clones to dominate HSC niches throughout the body, so what we learn about normal HSC development and homing will also likely apply to a variety of disorders of hematopoiesis, including clonal hematopoiesis and leukemias. The experiments here concern HSC homing, anchoring, support, and niche competition. In Aim 1, we will utilize long-term (LT)-HSC reporter mice to explore the specificity of the niche, identify the cell types that form direct contact with LT-HSC, and analyze the adhesion molecules, cytokines and chemokines responsible for HSC and niche cross-talk. In Aim 2 we propose to understand the role of HSC-surrounding BM cells—focusing on adjacent sinusoidal endothelial cells and the essential BM stromal subsets and their expressed factors. That builds on our discovery and characterization of the skeletal stem cells (SSC), which clonally generate bone, cartilage, and several distinct BM stromal cells that support hematopoiesis. We plan to produce mice in which each cell subset in the niche, including HSC, can be engineered to knock out identified genes to decipher the complexity of niche-HSC interactions. This could lead to an understanding if there are means to increase the number of functional niches, whether modulation of the molecular interactions could clear the niche specifically to promote the HSC engraftment, and to provide targets for modulating clonal HSC expansions. !

项目概要 我们已经证明，我们首先分离的纯化造血干细胞 (HSC) 可以进行同种异体移植 没有 GvH（它们缺乏 T 细胞），阻止自身免疫 1 型糖尿病（和系统性红斑狼疮），以及 诱导对 HSC 供体组织/器官的移植耐受。然而，纯化的 HSC 移植不被使用， 部分原因是预处理方案的毒性。因此，提高我们的认识至关重要 HSC 如何与其本地支持环境（造血生态位）相互作用。本研究的目标 是为了进一步鉴定和表征骨髓 (BM) HSC 的细胞和分子成分 利基，并了解 HSC 与其利基之间持续的细胞间通讯是如何进行的 调节它们在健康和血液疾病中的归巢、锚定、生存和功能。在以前的版本中 这笔赠款我们建立了抗体来代替毒性疗法来调节接受者，并将进一步修改 改善植入和加速重建的方案。进一步了解 HSC 利基 相互作用对我们在病理学中阐明的干细胞竞争有影响，例如， 造血系统疾病，例如骨髓增生异常综合征 (MDS)、骨髓增生性肿瘤 (MPN) 和 突变克隆的扩增以产生克隆性造血，其中一些会引起急性髓系白血病 （反洗钱）。在白血病和骨髓增生异常综合征 (MDS) 的发展过程中，具有癌前突变的 HSC 克隆在竞争中胜过正常细胞 HSC，但 HSC 总数似乎没有变化，这意味着 HSC 数量受到利基的控制。 HSC 的自然再循环使得高度竞争的克隆能够在整个生命周期中主导 HSC 生态位。 因此我们对正常 HSC 发育和归巢的了解也可能适用于各种 造血障碍，包括克隆造血和白血病。这里的实验涉及 HSC 归巢、锚定、支持和利基竞争。在目标 1 中，我们将利用长期 (LT)-HSC 报告基因 小鼠探索生态位的特异性，识别与 LT-HSC 直接接触的细胞类型，以及 分析负责 HSC 和生态位串扰的粘附分子、细胞因子和趋化因子。瞄准 2 我们建议了解 HSC 周围 BM 细胞的作用——关注相邻的正弦曲线 内皮细胞和必需的 BM 基质亚群及其表达因子。这建立在我们的 骨骼干细胞 (SSC) 的发现和表征，该细胞可克隆产生骨、软骨和 几种支持造血功能的不同骨髓基质细胞。我们计划培育出每个细胞都 生态位中的子集，包括 HSC，可以被设计来敲除已识别的基因以破译复杂性 利基-HSC 相互作用。这可能会导致人们了解是否有办法增加 功能性生态位，分子相互作用的调节是否可以专门清除该生态位 促进 HSC 植入，并为调节克隆 HSC 扩增提供目标。 ！