Thrombus Formation In Vivo

描述(由申请人提供)： 这项名为“体内血栓形成”的计划项目资助，吸引了四位主要研究人员来关注血栓形成的过程。该小组率先开发了高速活体共聚焦显微镜和宽视场显微镜，利用激光诱导血栓形成模型研究活体小鼠血栓的形成。利用这项技术，他们将解决与血栓形成的三个阶段相关的一系列关键问题：启动、发展和繁殖。在题为组织因子和血栓形成的第一个项目中，Bruce Furie博士将研究组织因子的生理作用和组织因子激活的分子基础。他将研究培养的内皮细胞和活体小鼠血栓形成过程中组织因子形式的表达动力学。血栓形成过程中所需的组织因子的细胞来源将使用嵌合小鼠、黑手党小鼠和内皮细胞、血小板、髓系细胞或平滑肌细胞中缺乏组织因子的转基因小鼠进行评估。在题为血小板蛋白棕榈酰化和血栓形成的项目#2中，Robert Flaumenhaft博士将使用小鼠血栓形成模型评估蛋白质棕榈酰化在血小板激活和血栓形成中的作用，并确定哪些蛋白质经历翻译后棕榈酰化。在题为CD39及其在血栓形成中的作用的项目#3中，Simon Robson博士将测试这一假设，即CD39/NTPDase-1的表达调节血管系统和血小板激活中的核苷酸介导的信号，从而调节活体小鼠血栓形成过程中的血栓传播。在题为初始血小板黏附和血栓传播的项目#4中，Barbara Furie博士将研究GPVI/PAR4和PAR4/von Willebrand因子在血栓形成过程中的血小板激活中的组合作用，von Willebrand因子的细胞来源对血栓形成的贡献，以及凝血因子XII和凝血因子IX在组织因子介导的血栓生长中的作用。行政、活体显微镜和质谱学核心支持四个项目的活动。研究活体动物血栓形成的生物化学和细胞生物学的能力为了解血栓形成和制定预防血栓形成的策略提供了机会，血栓形成是西方社会发病率和死亡率的主要原因。

DESCRIPTION (provided by applicant): This Program Project Grant, entitled "THROMBUS FORMATION IN VIVO" brings four principal investigators to focus on the process of thrombogenesis. This group has pioneered the development of high speed intravital confocal and widefield microscopy to study thrombus formation in a living mouse using the laser induced thrombosis model. Using this technology, they will address a series of critical questions related to the three phases of thrombus formation: initiation, development and propagation. In Project #1 entitled TISSUE FACTOR AND THE INITIATION OF THROMBUS FORMATION, Dr. Bruce Furie will study the physiologic role of tissue factor and the molecular basis of tissue factor activation. He will examine the kinetics of expression of tissue factor forms in cultured endothelial cells and during thrombus formation in a living mouse. The cellular source of tissue factor required during thrombus formation will be evaluated using chimeric mice, MAFIA mice, and genetically altered mice that are deficient in tissue factor in endothelial cells, platelets, myeloid cells or smooth muscle cells. In Project #2 entitled PLATELET PROTEIN PALMITOYLATION AND THROMBUS FORMATION, Dr. Robert Flaumenhaft will evaluate the role of protein palmitoylation in platelet activation and thrombus formation using the mouse thrombosis model, and determine which proteins undergo posttranslational palmitoylation. In Project #3 entitled CD39 AND ITS ROLE IN THROMBOGENESIS, Dr. Simon Robson will test the hypothesis that modulated CD39/NTPDase-1 expression regulates nucleotide-mediated signaling in the vasculature and platelet activation, thus regulating thrombus propagation during thrombus formation in living mice. In Project #4 entitled INITIAL PLATELET ADHESION AND THROMBUS PROPAGATION, Dr. Barbara Furie will study the combined roles of GPVI/PAR4 and PAR4/von Willebrand factor in platelet activation during thrombus formation, the contribution of cellular sources of von Willebrand factor to thrombus development and the role of Factor XII and Factor IX in tissue factor-mediated thrombus propagation. Administrative, Intravital Microscopy and Mass Spectrometry cores support the activities of the four projects. The ability to study the biochemistry and cell biology of thrombus formation in living animals offers opportunities to understand thrombosis and develop strategies for preventing thrombosis, the major cause of morbidity and mortality in Western society.