Tissue Factor the Vascular Proteome and Thrombosis

该项目解决了血栓形成途径中尚未解决的问题。首先，血管内皮表面微域背景在组织因子（TF）与其底物因子X和IX的关联中是否起作用？这个问题正在以一种新的方式解决，即创建各种选择性血管血栓原，这些血栓原可以停靠在不同的内皮表面环境中，并且可以通过实时活体显微镜在体内观察，也可以在体外细胞测定和TF驱动血栓形成和弥散性血管内凝血的动物模型中观察。结合先进的分子水平成像方法，使用转基因小鼠，并与项目2合作探索细胞信号传导的差异，与项目3合作探索各种转基因小鼠（包括高表达人类TF的小鼠）的背景，与项目4合作接近完整的三维蛋白质相互作用以及预测的原子水平相互作用，新的数据和见解将会到来。推进初步发现一个小的约束肽基配体，用于参与因子X与功能性TF: via关联的关键相互作用蛋白表面位点，我们将能够首次选择性地抑制凝血/血栓形成的外源性肢体

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.