Neutrophils and Vascular Endothelial Permeability

中性粒细胞和血管内皮通透性

• 批准号: 7278917
• 负责人: Sean P Colgan
• 金额: $ 7.5万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7278917
• 关键词: CD antigens; Chordata; actin binding protein; biological signal transduction; cell migration; clinical research; cytoskeleton; glutamate receptor; glutamates; human tissue; hypoxia; immunoprecipitation; laboratory mouse; membrane proteins; molecular pathology; neutrophil; nucleotidases; protein purification; protein structure function; tissue /cell culture; vascular endothelium; vascular endothelium permeability; western blottings

DESCRIPTION (provided by applicant): During episodes of inflammation and hypoxia, vascular endothelial cells interact with polymorphonuclear leukocytes (PMN, neutrophils) in both direct and indirect ways. Under such conditions, PMN may significantly modulate endothelial function, including vascular barrier function. Our previous studies explored the possibility that activated PMN may liberate factors which promote endothelial barrier function, and as such, may serve as a counter-regulatory pathway for potentially detrimental factors liberated at such sites. These studies identified a number of previously unappreciated molecules important in endothelial permeability, including soluble mediators (e.g. 5'AMP, glutamate), membrane proteins (CD73, metabotropic glutamate receptors, alpha-v beta-3), and cytoskeletal crosslinking proteins [e.g. vasodilator-stimulated phosphoprotein (VASP)] in maintenance of junctional permeability during interactions with PMN. In this proposal, we will test the hypothesis that PMN-derived factors regulate endothelial permeability. As such, three specific aims are proposed to test this hypothesis. First, we will elucidate the molecular pathway(s) ecto-nucleotidase regulation of endothelial permeability. We will utilize extensive in vitro and in vivo models to answer basic questions regarding the cause and effect relationship between ecto-nucleotidase expression and vascular barrier function. Second, we will explore the molecular mechanisms of vascular barrier regulation by exogenous glutamate. As such, we will elucidate the cooperative contributions of endothelial glutamate receptors with VASP in vitro and in vivo. Third, we will determine the relationship between PMN-derived extracellular mediators with endothelial cell-cell junction coupling. In particular, we will concentrate efforts on understanding VASP interactions with endothelial tight junctions and the influence of glutamate and adenine nucleotides on junctional reorganization in real- time. The overall aim of this proposal is to elucidate the molecular pathways critical to vascular barrier regulation in hypoxia and during inflammation.

描述（由申请方提供）：在炎症和缺氧发作期间，血管内皮细胞以直接和间接方式与多形核白细胞（PMN，中性粒细胞）相互作用。在这种情况下，PMN可以显著调节内皮功能，包括血管屏障功能。我们以前的研究探讨了激活的中性粒细胞可能释放促进内皮屏障功能的因子的可能性，因此，可能作为在这些网站释放的潜在有害因子的反调节途径。这些研究确定了许多以前未被认识到的在内皮通透性中重要的分子，包括可溶性介质（例如5 'AMP、谷氨酸）、膜蛋白（CD 73、代谢型谷氨酸受体、α-v β-3）和细胞骨架交联蛋白[例如血管舒张剂刺激的磷蛋白（VASP）]，它们在与PMN相互作用期间维持连接通透性。在这个建议中，我们将测试的假设，中性粒细胞衍生的因子调节内皮通透性。因此，提出了三个具体目标来检验这一假设。首先，我们将阐明外核苷酸酶调节内皮通透性的分子途径。我们将利用广泛的体外和体内模型来回答有关外核苷酸酶表达和血管屏障功能之间的因果关系的基本问题。其次，我们将探讨外源性谷氨酸调节血管屏障的分子机制。因此，我们将阐明合作的贡献，血管内皮细胞谷氨酸受体与VASP在体外和体内。第三，我们将确定中性粒细胞衍生的细胞外介质与内皮细胞-细胞连接偶联之间的关系。特别是，我们将集中精力了解VASP与内皮紧密连接的相互作用，以及谷氨酸和腺嘌呤核苷酸对真实的连接重组的影响。该提案的总体目标是阐明缺氧和炎症过程中对血管屏障调节至关重要的分子途径。