CCR2 in Hematopoietic Stem Cell Homing, Macrophage Polarization and Organ Repair

CCR2 在造血干细胞归巢、巨噬细胞极化和器官修复中的作用

• 批准号: 8259745
• 负责人: ISRAEL F. CHARO
• 金额: $ 47.75万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8259745
• 关键词: Acetaminophen; Acute; Acute myocardial infarction; Atherosclerosis; Attention; Blood; Blood Cells; Blood Circulation; Bone Marrow; Bone Marrow Cells; Bone Marrow Stem Cell; CCL2 gene; CCL7 gene; CX3CL1 gene; Carbon Tetrachloride; Cardiac; Cell Lineage; Cell surface; Cells; Characteristics; Chemotaxis; Chronic; Commit; Data; EFRAC; Experimental Models; Fibrosis; Funding; Genes; Glean; Healed; Heart; Hematopoietic; Hematopoietic Stem Cell Mobilization; Hematopoietic stem cells; Hepatic; Hepatotoxicity; Homing; Human; Impairment; In Vitro; Infarction; Inflammation; Infusion procedures; Injury; Injury to Liver; Intravenous infusion procedures; Knock-in Mouse; Ligation; Liver; Liver Failure; Mediating; Medullary Hematopoiesis; Modeling; Mus; Myelogenous; Myocardial; Myocardial Infarction; Myocardial Ischemia; Natural regeneration; Organ; Peritoneum; Phenotype; Population; Proliferating; Proto-Oncogene Protein c-kit; Publishing; RNA; Recovery; Recovery of Function; Recruitment Activity; Resolution; Role; Signal Transduction; Site; Sorting - Cell Movement; Staining method; Stains; Stem cells; Testing; Therapeutic Uses; Thioglycolates; Tissues; Transfusion; Trees; United States National Institutes of Health; Wild Type Mouse; Work; chemokine receptor; healing; human disease; in vivo; injured; insight; liver function; macrophage; migration; monocyte; novel; progenitor; public health relevance; repaired; research study; restoration; stem; tissue regeneration; tissue repair; trafficking

DESCRIPTION (provided by applicant): Hematopoietic stem cells (HSCs) and progenitors (HPCs) are bone-marrow derived cells that give rise to terminally differentiated circulating blood cells. Recent work has implicated these cells in the repair of parenchymal tissue in the setting of inflammation, but the signals that regulate this trafficking are poorly understood. In other NIH funded worked we discovered and cloned CCR2, the chemokine receptor that regulates monocyte migration to MCP-1, and showed that CCR2-/- mice are protected in murine models of atherosclerosis. In preliminary, unpublished results we have now found that CCR2 is expressed on subsets of primitive HSCs as well as some myeloid HPCs. CCR2 mediates the chemotaxis of c-Kit+Lin- bone marrow derived cells to MCP-1 and MCP-3. Following instillation of thioglycollate WT, but not CCR2-/- stem cells were actively recruited to the peritoneum, and to the liver following administration of acetaminophen. Significantly, infusion of CCR2+/+, but not CCR2-/- HSCs/HPCs accelerated the resolution of liver damage, and the recruited cells expressed genes characteristic of the M2 macrophage phenotype. Building on these recent findings, we propose three interrelated specific aims to define the role of CCR2 in HSC/HPC trafficking, and their potential roles in the resolution of inflammation and injury. In Specific Aim 1 we will quantify expression of CCR2 on early hematopoietic stem cells. We will test the hypothesis that CCR2 is expressed on true, self-replicating stem cells, and mediates their chemotaxis to sites of tissue inflammation and injury. Using novel CCR2/RFP knock-in mice, we will quantify the expression of CCR2 on HSCs/HPCs in all branches of the hematopoietic tree in mice and determine whether activation of CCR2 mobilizes HSCs and HPCs from bone marrow. In Specific Aim 2 we will determine whether CCR2-mediated recruitment of HSCs or HPCs contributes to the resolution of inflammation and ischemic injury in models of acute (acetaminophen)- and chronic (carbon tetrachloride)-induced hepatotoxicity, will determine the fate of the recruited stem and progenitor cells, and will test the hypothesis that enriching HSCs/HPCs for those that express CCR2 will significanlty enhances tissue repair. In Specific Aim 3 we will turn our attention to a model of experimental myocardial infraction in mice, and will determine if CCR2+ or CCR2- HSCs/HPCs accelerate recovery of cardiac function, and whether CCR2 antagonists reduce inflammation and enhance functional recovery. Completion of the work described above will identify the signals regulating the homing of bone marrow stem and progenitor cells to injured tissue, and further our mechanistic understanding of their role in tissue repair and regeneration. PUBLIC HEALTH RELEVANCE: Work proposed in this application will help to identify a new and mechanisitic marker for hematopoietic (ie., bone marrow derived) stem cells. Transfusion of such cells has the potential to enhance treatment of important human diseases, such as liver failure and acute myocardial infarction. The preliminary results described in this application identify CCR2, a well-characterized chemokine receptor, as being critical for the mobilization of stem cells to sites of inflammation and injury. Completion of the Specific Aims of this application will greatly accelerate efforts to optimize the therapeutic use of bone marrow stem cells.

描述（由申请人提供）：造血干细胞（hsc）和祖细胞（HPCs）是骨髓来源的细胞，可产生终末分化的循环血细胞。最近的研究表明，在炎症的情况下，这些细胞参与了实质组织的修复，但调控这种运输的信号却知之甚少。在NIH资助的其他工作中，我们发现并克隆了CCR2，一种调节单核细胞向MCP-1迁移的趋化因子受体，并表明CCR2-/-小鼠在动脉粥样硬化小鼠模型中受到保护。在初步的未发表的结果中，我们发现CCR2在原始造血干细胞亚群以及一些髓系造血干细胞上表达。CCR2介导c-Kit+Lin-骨髓源性细胞对MCP-1和MCP-3的趋化性。在注射巯基乙酸后，WT，而不是CCR2-/-干细胞被积极招募到腹膜，在给药后被招募到肝脏。值得注意的是，输注CCR2+/+而非CCR2-/- hsc /HPCs加速了肝损伤的消退，募集的细胞表达了M2巨噬细胞表型特征的基因。基于这些最近的发现，我们提出了三个相互关联的具体目标，以确定CCR2在HSC/HPC运输中的作用，以及它们在炎症和损伤解决中的潜在作用。在特异性目标1中，我们将量化CCR2在早期造血干细胞中的表达。我们将验证CCR2在真实的、自我复制的干细胞上表达，并介导其趋化到组织炎症和损伤部位的假设。使用新型CCR2/RFP敲入小鼠，我们将量化CCR2在小鼠造血树所有分支的造血干细胞/造血干细胞上的表达，并确定CCR2的激活是否能从骨髓中动员造血干细胞和造血干细胞。在Specific Aim 2中，我们将确定CCR2介导的hsc或HPCs的募集是否有助于急性（对乙酰氨基酚）和慢性（四氯化碳）诱导的肝毒性模型中炎症和缺血性损伤的解决，将决定募集的干细胞和祖细胞的命运，并将检验表达CCR2的hsc /HPCs的富集将显著增强组织修复的假设。在Specific Aim 3中，我们将把注意力转向实验性小鼠心肌梗死模型，并将确定CCR2+或CCR2- hsc /HPCs是否加速心功能的恢复，以及CCR2拮抗剂是否减少炎症和增强功能恢复。上述工作的完成将确定调节骨髓干细胞和祖细胞归巢到损伤组织的信号，并进一步了解它们在组织修复和再生中的作用机制。