The bone marrow extracellular matrix: scaffold of hematopoiesis

骨髓细胞外基质：造血支架

• 批准号: 10861385
• 负责人: Shinobu Matsuura
• 金额: $ 8.7万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10861385
• 关键词: 3-Dimensional; Ablation; Address; Adhesives; Affect; Allogenic; Animals; Antibodies; Automobile Driving; Award; Blocking Antibodies; Blood; Blood Cells; Blood Vessels; Bone Marrow; Bone Marrow Diseases; Cell Proliferation; Cell secretion; Cell-Matrix Junction; Cells; Complementary therapies; Complex; Dependence; Development; Disease; Disease Outcome; Dysmyelopoietic Syndromes; Endothelial Cells; Environment; Exons; Extracellular Matrix; Failure; Fiber; Foundations; Gene Mutation; Genes; Genetic; Genomics; Goals; Hematology; Hematopoiesis; Hematopoietic; Human; Integrins; JAK2 gene; K-Series Research Career Programs; Life; Ligands; Lysine; Malignant - descriptor; Mass Spectrum Analysis; Mediating; Mentorship; Microanatomy; Modeling; Mouse Strains; Mus; Mutation; Mutation Analysis; Myelofibrosis; Outcome; Pathogenesis; Pathology; Patients; Pericytes; Play; Post Translational Modification Analysis; Post-Translational Protein Processing; Primary Myelofibrosis; Procedures; Production; Prognosis; Proliferating; Protein-Lysine 6-Oxidase; Proteins; Proteomics; Publishing; Reporting; Research; Research Personnel; Sampling; Signal Transduction; Spatial Distribution; Stem cell transplant; Stromal Cells; Survival Rate; Techniques; Testing; Therapeutic; Tissues; Training; Transgenes; Transgenic Mice; Uterus; career; confocal imaging; curative treatments; defined contribution; effective therapy; experience; high risk; human disease; human model; improved; in vivo; inhibitor; leukemia; mortality; mouse model; mutant; novel; programs; promoter; scaffold

ABSTRACT This proposal addresses the urgent need to understand disease mechanisms in Primary Myelofibrosis (PMF). PMF is a disease of dismal prognosis. The only potentially curative therapy, allogeneic stem cell transplantation, is a high-risk procedure, with a mortality rate of at least 50%. Myelofibrosis is characterized by excessive production of extracellular matrix (ECM), the fibers in bone marrow, which leads to a progressive failure in blood cell production. A number of gene mutations, including JAK2V617F, have been reported in PMF. While it is thought that hematopoietic cells carrying gene mutations abnormally proliferate and secrete factors that stimulate stromal cells to produce ECM excessively, inhibition of JAK2 was not able to improve disease outcomes, and complementary therapies that specifically target ECM production are not currently available. Dependence on the prevalent hypothesis, with its focus on gene mutations, is a critical barrier to progress in the field. This project proposes, instead, to address the problem from the ECM angle. Building on our published and preliminary studies with mouse and human samples, the central hypothesis of this proposal, based is that abnormal activation of integrins (adhesive molecules that mediate cell attachment to ECM) facilitates proliferation of PMF cells (Aim 1A-C). Further, I will explore two potential mechanisms behind this abnormal integrin activation, asking, first, whether abnormally high ECM production occurs in the vascular niche, where PMF cells develop (Aim 1D), and, secondly, whether lysyl oxidase (LOX), a protein upregulated in PMF, activates integrins in PMF through post-translational modification (Aim 2). As PMF proliferation arises from interaction between ECM and integrins, the question is whether proliferation is triggered by an abnormality in ECM formation or by direct integrin activation by LOX. These studies will be performed on the most representative mouse model of human PMF, mice transgenic for the JAK2V617F mutation. In Aim 1A-C the effect of integrins on cell proliferation in PMF will be determined by blocking integrin function with antibodies and by deleting integrin genes using genetically modified mouse strains. In Aim 1D, 3D confocal imaging will be used to determine the spatial distribution of cells, ECM and vasculature in myelofibrotic bone marrow. In Aim 2, proteomic analysis of post-translational modifications will be performed in integrins to determine specific lysine residues oxidized by LOX. This is a Career Development Award, and the PI of this application will have the mentorship of investigators with expertise in the field for the execution of the project. The project builds on previous experience of the PI in malignant hematology. The training and the expertise acquired during this award will provide a foundation for a successful independent career for the PI of this project.

摘要 这项建议解决了了解原发性骨髓纤维化(PMF)疾病机制的迫切需要。 PMF是一种预后不佳的疾病。唯一有可能治愈的疗法，异基因干细胞 移植，是一种高风险的手术，死亡率至少为50%。骨髓纤维化的特征是 过度产生细胞外基质(ECM)，这是骨髓中的纤维，导致进行性 血细胞生产失败。一些基因突变，包括JAK2V617F，已经在 PMF。而携带基因突变的造血细胞被认为是异常增殖和分泌 过度刺激基质细胞产生ECM的因素，对JAK2的抑制不能改善 疾病结果，以及专门针对ECM产生的补充疗法目前还没有 可用。对流行假说的依赖，其重点是基因突变，是 这一领域的进步。相反，该项目建议从ECM的角度来解决这一问题。在基础上建设 我们发表的对老鼠和人类样本的初步研究，这一提议的中心假设， 基础是整合素(介导细胞与细胞外基质附着的黏附分子)的异常激活 促进PMF细胞的增殖(目标1A-C)。此外，我将探索这背后的两个潜在机制 异常的整合素激活，首先，询问血管中是否出现异常高的ECM产生 第二，赖氨酰氧化酶(LOX)，一种蛋白质是否上调 在PMF中，通过翻译后修饰激活PMF中的整合素(目标2)。随着PMF扩散的出现 从ECM和整合素之间的相互作用来看，问题是增殖是否由一种 细胞外基质形成异常或LOX直接激活整合素。这些研究将在 最具代表性的人PMF小鼠模型，转基因小鼠为JAK2V617F突变。在目标1A-C中 整合素对PMF细胞增殖的影响将通过阻断整合素的功能来确定。 通过使用转基因小鼠品系来删除整合素基因。在AIM 1D、3D共焦中 成像将用于确定骨髓纤维病骨中细胞、细胞外基质和血管的空间分布。 骨髓。在目标2中，将对整合素进行翻译后修饰的蛋白质组学分析，以 测定LOX氧化的特定赖氨酸残基。这是一个职业发展奖，而这个的PI 应用程序将得到具有该领域专业知识的调查人员的指导，以执行该项目。 该项目建立在恶性血液病PI以前的经验基础上。培训和专业知识 在此奖项期间获得的将为该公司的PI成功的独立职业生涯奠定基础 项目。