Molecular and Cellular Biology of Megakaryocytes in Normal and Neoplastic Hematopoiesis

正常和肿瘤造血中巨核细胞的分子和细胞生物学

• 批准号: 9216195
• 负责人: Huichun Zhan
• 金额: $ 35.67万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-15 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9216195
• 关键词: Abnormal megakaryocyte; Adhesives; Affect; Angiogenic Factor; Antibodies; Biological Assay; Blood Cells; Blood Platelets; Blood Vessels; Cell Adhesion Molecules; Cell Count; Cell Cycle; Cell Line; Cell Proliferation; Cell physiology; Cells; Clonal Expansion; Complex; Disease; Effectiveness; Endothelial Cells; Extracellular Matrix; Fluorescence Microscopy; Genetic; Goals; Growth; Hematologic Neoplasms; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Human; Hyperplasia; Immunofluorescence Immunologic; In Vitro; Intracellular Membranes; Maps; Marrow; Megakaryocytes; Membrane Proteins; Methods; Molecular; Molecular and Cellular Biology; Mus; Mutation; Myeloproliferation; Myeloproliferative disease; Neoplasms in Vascular Tissue; PTPRC gene; Pathway interactions; Patients; Physiological; Play; Proteins; Proteomics; Regulation; Research Proposals; Role; Signal Transduction; Signaling Molecule; Source; Staining method; Stains; Stem cells; Syndrome; Technology; Thrombopoietin; Transgenic Organisms; Treatment Efficacy; Umbilical vein; Vascular Endothelial Cell; Work; angiogenesis; base; cell growth; clinical phenotype; cytokine; density; in vivo; induced pluripotent stem cell; inhibitor/antagonist; knockout gene; mouse model; mutant; neoplastic; receptor; small hairpin RNA

PROJECT SUMMARY/ABSTRACT Megakaryocytes (MK) are rare very large marrow cells that give rise to blood platelets. Recent evidence has implicated MKs in regulating hematopoietic stem/progenitor cell (HSPC) activity by the many cytokines and extracellular matrix components produced by these cells. Many hematologic malignancies are associated with MK abnormalities. The myeloproliferative neoplasms (MPNs) are clonal stem cell disorders characterized by HSPC expansion and overproduction of mature blood cells. The acquired mutation JAK2V617F plays a central role in these disorders, but the precise molecular mechanisms responsible for MPN HSPC expansion are not fully understood, limiting the effectiveness of current treatments. MK hyperplasia is a hallmark feature of MPNs. We have shown that JAK2V617F-bearing MKs cause a murine myeloproliferative syndrome with HSPC expansion and increased marrow sinusoidal vascular density. In addition, we have demonstrated that the MPN vascular niche is important for JAK2V617F HSPC clonal expansion. Based on these initial results, we hypothesize that JAK2V617F-bearing MKs have altered hematopoietic niche function which not only enhances stem cell function directly but also affects the vascular niche to indirectly promote HSPC expansion in MPNs. The objective of the proposed work is to determine the physiological effects and the molecular mechanism(s) by which JAK2V617F-bearing MKs maintain and expand HSPCs in MPNs. In particular, we propose the following three specific aims: 1) To study the effects of JAK2V617F-bearing MKs on stem cell function using both a transgenic murine model and human induced pluripotent stem cell lines; 2) To study the effects of JAK2V617F MKs on endothelial cell function in vitro, and the vascular niche and HSPC expansion in vivo. The role of thrombopoietin/MPL signaling in JAK2V617F MK niche function will be assessed; 3) To systemically investigate how JAK2V617F mutation affects megakaryocyte intracellular, membrane, and secreted proteins by quantitative proteomics. The function of selected proteins in HSPC function and vascular niche function will be explored. The long term goal of this research proposal is to identify the complex network of MK signaling in both normal and neoplastic hematopoiesis and develop additional, more effective therapeutic strategies in MPNs and potentially other hematologic malignancies.

项目摘要/摘要 巨核细胞(MK)是一种罕见的能产生血小板的非常大的骨髓细胞。最近的证据表明 MKs通过多种细胞因子和细胞因子调节造血干/祖细胞的活性 由这些细胞产生的细胞外基质成分。许多恶性血液病与 MK异常。骨髓增生性肿瘤(MPN)是克隆性干细胞疾病，其特征是 HSPC的扩增和成熟血细胞的过度生产。获得性突变JAK2V617F在 在这些疾病中发挥作用，但导致MPN HSPC扩张的确切分子机制并不是 完全理解，限制了目前治疗的有效性。巨核细胞增生症是该病的一个显著特征 MPN。我们已经证明，携带JAK2V617F的MKs可引起小鼠单纯疱疹病毒携带者的骨髓增殖综合征。 扩张，骨髓窦血管密度增加。此外，我们还证明了MPN 血管生态位对JAK2V617F HSPC克隆扩增具有重要意义。基于这些初步结果，我们 假设携带JAK2V617F的MKs改变了造血龛功能，不仅增强了 干细胞功能直接也影响血管壁龛，间接促进HSPC在MPNS中的扩增。 拟议工作的目标是确定生理效应和分子机制(S) 携带JAK2V617F的MKs在MPN中维持和扩展HSPC。特别是，我们提出了以下建议 目的：1)研究JAK2V617F-MKs对干细胞功能的影响。 转基因小鼠模型和人诱导的多能干细胞系；2)研究JAK2V617F的作用 MKS对血管内皮细胞功能的影响，以及对血管壁龛和HSPC体内扩张的影响。的作用 将评估促血小板生成素/MPL信号在JAK2V617F中的MK生态位功能；3)系统研究 JAK2V617F突变对巨核细胞胞内、膜和分泌蛋白的影响 蛋白质组学。将探索所选蛋白质在HSPC功能和血管壁龛功能中的作用。 这项研究计划的长期目标是识别MK信令的复杂网络 和肿瘤造血，并开发额外的，更有效的治疗MPNS和 可能是其他恶性血液病。