Regulation of Endothelial Cell Procoagulant Properties

内皮细胞促凝血特性的调节

• 批准号: 7029344
• 负责人: WOLFRAM RUF
• 金额: $ 43.09万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7029344
• 关键词: atherosclerosis; atherosclerotic plaque; blood coagulation; clinical research; hemostasis; integrins; laboratory mouse; laboratory rabbit; oxidation reduction reaction; pathologic process; thromboplastin; thrombosis; vascular endothelium

Tissue Factor is the central activator of the coagulation cascade and has emerged as a signaling receptor at the crossroads of G-protein coupled and integrin signaling pathways. Project 3 is based on our recent finding that reduction/oxidation of a critical disulfide bond in the extracellular domain of TF regulates prothrombotic activity. To elucidate how this novel regulatory mechanism separately controls signaling and prothrombotic activity of TF is the central theme of this project. Specifically, the applicant proposes to characterize the protein structural determinants that render TF susceptible to reduction in the context of enzyme and substrate interaction, the role of membrane composition on the stability of the critical disulfide bond, and the localization of reduced TF relative to cellular oxidative pathways, including protein disulfide isomerase implicated in oxidative activation of TF. The finding that TF can alternatively become S-nitrosylated is the basis for experiments to characterize cellular pathways that transfer NO to TF and to generate analytical tools to detect this posttranslation modification in the context of thrombus formation. The applicant proposes to test the hypothesis that thrombogenicity is under reciprocal control by signaling pathways with which TF is associated. Specifically, strategies are developed to demonstrate that integrins target TF to extracellular matrices and that the effects of integrin activation on local redox potential contribute to matrix thrombogenicty by oxidating TF. Mutagenesis is employed to unambiguously establish the signaling properties of the non-coagulant form of TF. Thrombogenic and signaling roles of TF are evaluated in atherosclerosis using an in vivo model of turbulent flow induced lesion development in atherosclerosis prone animals. The successful completion of these studies will provide valuable information on a novel regulatory pathway of TF-dependent thrombogenicity of particular relevance for the dynamics of platelet activation on extracellular matrices.

组织因子是凝血级联反应的中心激活剂，并已作为信号受体出现， G蛋白偶联和整合素信号通路的十字路口。项目3是基于我们最近的发现 TF细胞外结构域中一个关键二硫键的还原/氧化调节血栓前 活动为了阐明这种新的调节机制如何分别控制信号传导和血栓前 TF的活动是这个项目的中心主题。具体而言，申请人建议表征 蛋白质结构决定因素，使TF易于减少的情况下，酶和 底物相互作用，膜组成对关键二硫键稳定性的作用，以及 相对于细胞氧化途径的还原型TF的定位，包括蛋白质二硫键异构酶 与TF的氧化激活有关。TF可以替代地变成S-亚硝基化的发现是 实验的基础，以表征将NO转移到TF的细胞途径，并产生分析 在血栓形成的背景下检测这种翻译后修饰的工具。申请人建议 为了检验血栓形成性受TF信号通路相互控制的假设， 关联的.具体地说，开发策略来证明整合素靶向TF到细胞外， 整合素活化对局部氧化还原电位的影响有助于基质 氧化TF的血栓形成作用。采用诱变来明确地建立信号传导 TF的非凝结形式的性质。TF的血栓形成和信号传导作用在 使用动脉粥样硬化倾向中湍流诱导病变发展的体内模型的动脉粥样硬化 动物这些研究的成功完成将为一种新的监管机制提供有价值的信息。 TF依赖性血栓形成途径与血小板活化动力学特别相关， 细胞外基质