The bone marrow extracellular matrix: scaffold of hematopoiesis

骨髓细胞外基质：造血支架

• 批准号: 10576364
• 负责人: Shinobu Matsuura
• 金额: $ 9.92万
• 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/10576364
• 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; CRISPR/Cas technology; Cell Proliferation; Cell secretion; Cell-Matrix Junction; Cells; Complementary therapies; Complex; Computer Analysis; Dependence; Development; Disease; Disease Outcome; Dysmyelopoietic Syndromes; Endothelial Cells; Environment; Enzymes; 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; Morphology; 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; 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; targeted treatment; therapy development

ABSTRACT This proposal addresses the urgent need to understand disease mechanisms in Primary Myelofibrosis (PMF). PMF is a disease with a 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 only partially improves 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 is that abnormal activation of integrins (adhesive molecules that mediate cell attachment to ECM) facilitates proliferation of PMF cells. 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. Thus, I will explore two potential mechanisms behind this abnormal integrin activation, asking whether: (1) abnormally high ECM production occurs in the vascular niche, where PMF cells develop, and (2) Lysyl oxidase (LOX), an ECM enzyme upregulated in PMF, activates integrins in PMF through post-translational modification. These studies will be performed on the most representative mouse model of human PMF, mice transgenic for the JAK2V617F mutation. Effects of integrins on cell proliferation in PMF will be determined by blocking integrin function with antibodies and by deleting integrin genes using CRISPR/Cas9. I will use 3D confocal imaging and advanced computational analysis to determine the spatial distribution of cells, ECM, and vasculature in myelofibrotic bone marrow, clarifying the mechanisms of ECM production by detailed morphological analysis of the vascular niche. Proteomic analysis of post-translational modifications will be performed on integrins to determine specific lysine residues oxidized by LOX, which has implications on the development of therapies targeting integrins or 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.

抽象的 该提议旨在了解原发性骨髓纤维化（PMF）中疾病机制的迫切需求。 PMF是一种疾病，预后很沮丧。唯一的潜在治疗疗法，同种异体干细胞 移植是一种高风险程序，死亡率至少为50％。骨髓纤维化的特征是 细胞外基质（ECM）的过度产生，骨髓中的纤维，导致渐进性 血细胞产生失败。已经报道了许多基因突变，包括JAK2V617F PMF。虽然认为携带基因突变的造血细胞异常增殖并分泌 刺激基质细胞产生ECM过度产生ECM的因素，JAK2的抑制只会部分改善 目前，疾病结果和专门针对ECM的互补疗法目前尚未 可用的。对普遍假设的依赖，其侧重于基因突变，这是通往基因突变的关键障碍 在现场进展。该项目建议从ECM角度解决问题。建立 我们使用小鼠和人类样本的发表和初步研究，该提议的中心假设 整联蛋白的异常激活（介导细胞附着到ECM的粘合分子）有助于促进 PMF细胞的增殖。由于PMF的增殖来自ECM和整合素之间的相互作用，因此 问题是增殖是由ECM形成异常还是由直接整联蛋白激活触发的。 因此，我将探索这种异常整联蛋白激活背后的两个潜在机制，询问：（1） ECM异常高的生产发生在PMF细胞中的血管生态位，（2）赖氨酸氧化酶 （LOX）是一种上升在PMF中的ECM酶，通过翻译后修饰激活PMF中的整联蛋白。 这些研究将在人类PMF的最具代表性的小鼠模型，小鼠转基因模型上进行 JAK2V617F突变。整联蛋白对PMF细胞增殖的影响将通过阻塞来确定 通过抗体和使用CRISPR/CAS9删除整联蛋白基因的整合素功能。我将使用3D共享 成像和高级计算分析，以确定细胞，ECM和 骨髓纤维化骨髓中的脉管系统，通过详细说明ECM生产的机制 血管生态位的形态分析。翻译后修饰的蛋白质组学分析将是 在整联蛋白上执行以确定由LOX氧化的特定赖氨酸残基，这对 靶向整联蛋白或LOX的疗法的开发。这是一个职业发展奖，也是这样的PI 申请将在该领域内具有专业知识的调查人员的指导，以执行该项目。 该项目以恶性血液学PI的先前经验为基础。培训和专业知识 在此奖项期间获得的将为成功的PI的成功独立职业提供基础 项目。