PARACRINE REGULATION OF ANGIOGENESIS BY MURAL CELLS

壁细胞对血管生成的旁分泌调节

基本信息

批准号：
7568229
负责人：
ROBERTO NICOSIA
金额：
$ 31.5万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-04-01 至 2011-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7568229
关键词：
Adult Angiopoietin-1 Atherosclerosis Binding Biochemical Biological Assay Blood Vessels CD14 Antigen CD14 gene Cancer Patient Cardiovascular system Cell Separation Cells Confocal Microscopy Coronary artery Dependency Disease Dose Endothelial Cells Enzyme-Linked Immunosorbent Assay Failure Gene Expression Regulation Goals Grant Immune Immune response Immune system In Vitro Injury Knowledge Lasers Life Ligands MAPK14 gene Malignant Neoplasms Mediating Methods Microarray Analysis Microscopy Modeling Molecular Monocyte Chemoattractant Protein-1 Mus Organ Organ Culture Techniques Pathway interactions Pattern Peripheral Vascular Diseases Play Process Production Rattus Receptor Protein-Tyrosine Kinases Recombinants Regulation Research Personnel Reverse Transcriptase Polymerase Chain Reaction Role Signal Pathway Signaling Molecule System TIE-2 Receptor TLR4 gene Time Toll-Like Receptor 2 Transgenic Mice Transgenic Organisms Tumor Necrosis Factor-alpha Tumor Necrosis Factors Up-Regulation Vascular Endothelial Growth Factors Work Wound Healing angiogenesis base cell motility chemokine cytokine design human TNFRSF1A protein immunocytochemistry in vivo Model insight macrophage neutralizing antibody novel paracrine precursor cell research study response response to injury therapeutic angiogenesis therapeutic development tumor necrosis factor alpha receptor tumor progression

项目摘要

DESCRIPTION (provided by applicant): Angiogenesis, the formation of new blood vessels by endothelial cells, plays a critical role in wound healing, the revascularization of ischemic organs, and the progression of atherosclerosis and cancer. The basic theme of this grant is that periendothelial mural cells regulate angiogenesis by paracrine mechanisms. Our previous work focused on the role of the mural cell-derived product angiopoietin-1 (Ang-1) and its receptor Tie2 in the aortic ring model of injury-induced angiogenesis. The following novel findings emerged from these studies. 1. Ang-1 induces expression of tumor necrosis factor-alpha (TNF) and other immune-related cytokines/chemokines, which stimulate macrophage recruitment and angiogenesis. 2. TNF is required for the angiogenic activity of Ang-1-induced cytokines. 3. Ang-1-mediated mural cell recruitment can be inhibited by blocking the p38 MARK pathway, which regulates production of monocyte chemotactic protein-1 (MCP-1), a mural cell stimulatory chemokine induced by Ang-1. 4. Ang-1 promotes expression of Toll-like receptor-2 (TLR2) and the TLR4 adapter molecule CD14, which regulate cytokine production in the innate immune system. 5. Stimulation of TLR2 with specific ligands dose-dependently promotes angiogenesis. 6. The angiogenic response of the aortic wall is impaired in mice with disrupted MyD88, a key TLR signaling molecule. On this basis we formulated the hypothesis that Ang-1 regulates angiogenesis in response to injury by cooperating with the innate immune system. Specific aims: (1) To establish the role of vascular macrophages and their cytokine product TNF in Ang-1-mediated angiogenesis. (2) To define the role of MCP-1 in Ang-1/p38 MAPK-mediated mural cell recruitment during angiogenesis. (3) To determine the function of TLRs in the angiogenic response of the vessel wall. For these studies we will use rat and mouse in vitro and in vivo models of angiogenesis. Functional studies will be performed with neutralizing antibodies and genetically modified mice with defective TNF and TLR systems. Macrophages will be selectively ablated using biochemical and transgenic methods. Molecular studies will include real-time RT-PCR, ELISA, laser capture microscopy, and confocal microscopy. Proposed experiments will define new molecular mechanisms of angiogenesis and vessel wall assembly. This knowledge may provide novel insights for the development of therapeutic approaches aimed at treating angiogenesis-dependent disorders and ischemic conditions.

描述（由申请人提供）：血管生成，内皮细胞形成新血管，在伤口愈合，缺血器官的血运重建以及动脉粥样硬化和癌症的进展中起着至关重要的作用。这项赠款的基本主题是，旁皮细胞细胞通过旁分泌机制调节血管生成。我们以前的工作着重于壁细胞衍生的产物血管生成素-1（ANG-1）及其受体TIE2在损伤引起的血管生成的主动脉环模型中的作用。这些研究从以下新颖的发现中得出。 1。Ang-1诱导肿瘤坏死因子-Alpha（TNF）和其他与免疫相关的细胞因子/趋化因子的表达，这些因子刺激巨噬细胞募集和血管生成。 2。TNF是Ang-1诱导的细胞因子的血管生成活性所必需的。 3。Ang-1介导的壁画细胞募集可以通过阻断p38 MARK途径来抑制，p38 MARK途径可以调节单核细胞趋化蛋白-1（MCP-1）的产生，这是一种由ANG-1诱导的壁细胞细胞刺激趋化因子。 4。Ang-1促进了Toll样受体-2（TLR2）和TLR4适配器分子CD14的表达，这些分子CD14调节先天免疫系统中的细胞因子产生。 5。刺激特定配体剂量依赖性地促进血管生成的TLR2刺激。 6。主动脉壁的血管生成反应在MyD88（密钥TLR信号分子）的小鼠中受损。在此基础上，我们提出了以下假设，即Ang-1通过与先天免疫系统合作来调节造成损伤的血管生成。具体目的：（1）在ANG-1介导的血管生成中建立血管巨噬细胞及其细胞因子产物TNF的作用。（2）在血管生成过程中定义了MCP-1在ANG-1/p38 MAPK介导的壁细胞募集中的作用。（3）确定TLR在血管壁的血管生成反应中的功能。对于这些研究，我们将在体外和体内使用大鼠和小鼠的血管生成模型。功能研究将使用具有缺陷的TNF和TLR系统的中和抗体和转基因的小鼠进行。巨噬细胞将使用生化和转基因方法选择性地烧毁。分子研究将包括实时RT-PCR，ELISA，激光捕获显微镜和共聚焦显微镜。提出的实验将定义血管生成和血管壁组件的新分子机制。这些知识可能为开发旨在治疗血管生成依赖性疾病和缺血状况的治疗方法的发展提供新的见解。