Tissue Factor the Vascular Proteome and Thrombosis

组织因子、血管蛋白质组和血栓形成

• 批准号: 7237995
• 负责人: THOMAS Scott EDGINGTON
• 金额: $ 64.72万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7237995
• 关键词: X ray crystallography; blood coagulation; clotting factor; coagulation factor IX; coagulation factor VII; coagulation factor X; confocal scanning microscopy; enzyme activity; enzyme structure; intravital microscopy; laboratory mouse; laboratory rat; membrane structure; nuclear magnetic resonance spectroscopy; nucleic acid sequence; polymerase chain reaction; protease inhibitor; proteasome; protein structure function; proteomics; site directed mutagenesis; structural biology; thromboplastin; thrombosis; tissue /cell culture; vascular endothelium

This project addresses issues in the thrombogenic pathways that have yet to be resolved. First, is the role of vascular endothelial surface microdomain context in the association of Tissue Factor (TF) with its substrates factors X and IX? This is being addressed in a novel manner with creation of various Selective Vascular Thrombogens that dock to different endothelial surface context and can be observed in vivo by real-time intravital microscopy as well as in in vitro cell assays and animal models of TF driven thrombosis and disseminated intravascular coagulation. Coupled with advanced imaging methods at the molecular level, use of genetically modified mice, and the collaboration with Project 2 to explore the differences in cell signaling, with Project 3 to explore the context in various genetically modified mice including high human TF expressing mice, and with Project 4 to approach the full three dimensional protein interactions coupled with the predicted atomic level interactions, a new level of data and insight will be forthcoming. Advancing initial discovery of a small constrained peptidyl ligand for a key interactive protein surface locus involved in association of the factor X with the functional TF:VIIa we will be able for the first time to selectively inhibit the extrinsic limb of the coagulation/thrombogenic pathway without inhibiting factor IX activation and effecting the intrinsic limb of the coagulation hemostatic pathways. This will require collaboration with all projects and cores for its execution. This set of studies will also address the correct molecular identification by mass spectrometry of the real endothelial surface factor Xa specific receptor. This is currently in process and potential candidate molecules have been identified and bear no relationship to the current scientific content of the literature. How important this receptor, FXaR, may be in thrombogenic and hemostatic pathways will be explored as well as its structure and basis of function. We have established by independent search methods a set of 92 peptidyl probes selective for the arterial atherosclerotic plaque surface endothelium. We have completed identification of one of the targets and are advancing three others to protein identification. The initially solved target that is very selective in vivo in atherosclerotic ApoE knockout mice is also similarly expressed on the surface of human iliac artery atherosclerotic plaques ex vivo. We will assess the linkage between plaque expression of these panels of markers and the histology, human TF content in the huTF knockin mouse/TF knockout mice as well as explore foam cell content and other possible markers of thrombotic risk of plaques. Such data and the probes may provide the much lacking means of in vivo imaging of the vasculature of humans for identification of the high risk 'susceptible' plaques.

该项目解决了尚未解决的血栓形成途径中的问题。首先，血管内皮表面微域环境在组织因子（TF）与其底物因子X和IX的关联中的作用？这是一种新颖的方式解决的，创建了各种选择性的血管血栓形成，这些血栓形成了，这些血栓源是将其停靠在不同的内皮表面上下文中，并且可以通过实时的体内丢入式显微镜以及体外细胞分析和TF驱动势力势力势力促进性血栓形成和散发性内部肠内凝结的动物模型在体内观察到这一点。 Coupled with advanced imaging methods at the molecular level, use of genetically modified mice, and the collaboration with Project 2 to explore the differences in cell signaling, with Project 3 to explore the context in various genetically modified mice including high human TF expressing mice, and with Project 4 to approach the full three dimensional protein interactions coupled with the predicted atomic level interactions, a new level of data and insight will be forthcoming.推进对因子X与功能性TF关联的关键互动蛋白表面基因座的初步发现，该蛋白质表面与功能性TF：VIIA相关性：VIIA我们将首次能够选择性地抑制凝血/血栓形成的外部肢体 途径没有抑制因子IX激活并影响凝结的固有肢 止血途径。这将需要与所有项目和核心进行执行。这组研究还将通过对实际内皮表面因子XA特异性受体的质谱法进行正确的分子鉴定。目前正在进行过程中，并且已经确定了潜在的候选分子，并且与文献当前的科学内容没有任何关系。该受体FXAR在血栓形成和止血途径中的重要性以及其功能的结构和基础将被探索。我们通过独立搜索方法建立了一组92个肽基探针，可为动脉动脉粥样硬化斑块表面内皮选择性选择性。我们已经完成了对目标之一的识别，并将另外三个靶标介绍到蛋白质鉴定上。最初解决的靶标在动脉粥样硬化的APOE基因敲除小鼠中非常有选择性的靶标在人动脉动脉动脉粥样硬化斑块的表面上类似地表达。我们将评估这些标记面板的斑块表达与HUTF敲除小鼠/TF基因敲除小鼠中的人类TF含量以及探索泡沫细胞含量和其他其他组织学之间的联系 斑块的血栓形成风险的可能标记。这样的数据和探针可能会提供很多 缺乏人类脉管系统的体内成像用于识别高风险 “易感性”斑块。