Thrombin Induced Pulmonary Vascular Permeability

凝血酶诱导的肺血管通透性

• 批准号: 7067620
• 负责人: Asrar B. Malik
• 金额: $ 38.05万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-06-11 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7067620
• 关键词: G protein; angiogenesis; angiopoietins; biological signal transduction; cadherins; calcium flux; enzyme activity; genetically modified animals; laboratory mouse; lung; phosphorylation; protein kinase C; receptor binding; receptor expression; respiratory circulation; second messengers; thrombin; tight junctions; tissue /cell culture; transfection; vascular endothelium permeability

DESCRIPTION (provided by applicant): We will extend our studies of the cellular and molecular mechanisms that are responsible for increased permeability of the lung vasculature. This increase in lung vascular permeability results in an array of pathological conditions including Adult Respiratory Distress Syndrome. The studies proposed here are based on the hypothesis that the VEcadherin- containing adherens junctions (AJs) of endothelial cells are critical determinants in both the increased permeability response of the endothelium and its time-dependent reversal. The dynamic AJ affects cell shape through the interaction of VE-cadherin with catenins that in turn associate with the actin cytoskeleton. The proposed studies will characterize the mechanisms by which AJs disassemble in response to thrombin and subsequently reassemble in a time-dependent manner. We will identify the phosphorylation and dephosphorylation "switches", present in the VEcadherin/ eatenin complexes, which are responsible for regulating endothelial permeability. This project has the following Aims: (i) to identify the signaling mechanisms mediating the loss of homotypic VE-cadherin binding, the disassembly of AJs, and the resultant increased permeability of the endothelium, (ii), to characterize the signaling mechanisms responsible for re-assembly of AJs and reversal of this increased permeability response, and (iii) to characterize the mechanism of angiopoietin-1-induced AJ stabilization and its role in inhibiting the loss of endothelial barrier function. The studies will be carried out both in endothelial cells and in vivo with intact mice. We will exploit a variety of methodologies including cell imaging, expression of mutant constructs, gene transfer, and the evaluation of transgenic mouse models. By more fully understanding mechanisms of increased vascular permeability and the reversal of this process, we will be in a position to develop novel strategies directed against components of the AJ complex that will prevent vascular leakiness and the formation of pulmonary edema.

描述（由申请人提供）： 我们将扩展对导致肺脉管系统渗透性增加的细胞和分子机制的研究。肺血管通透性的增加导致一系列病理状况，包括成人呼吸窘迫综合征。这里提出的研究基于以下假设：在内皮细胞的内皮细胞中含有粘膜蛋白 - 含有粘附蛋白的连接（AJ）都是重要的决定因素，而内皮的渗透性响应及其时间依赖性逆转。动态AJ通过VE-钙粘着蛋白与Catenin的相互作用影响细胞形状，而Catenin又与肌动蛋白细胞骨架相关。拟议的研究将表征AJS响应凝血酶并随后以时间依赖性方式重新组装的机制。我们将确定存在于维卡德蛋白/ Eatenin络合物中的磷酸化和去磷酸化的“开关”，这些磷酸化和磷酸化蛋白复合物负责调节内皮渗透性。 This project has the following Aims: (i) to identify the signaling mechanisms mediating the loss of homotypic VE-cadherin binding, the disassembly of AJs, and the resultant increased permeability of the endothelium, (ii), to characterize the signaling mechanisms responsible for re-assembly of AJs and reversal of this increased permeability response, and (iii) to characterize the mechanism of血管生成素1诱导的AJ稳定及其在抑制内皮屏障功能丧失中的作用。这些研究将在内皮细胞和完整小鼠的体内进行。我们将利用各种方法，包括细胞成像，突变构建体的表达，基因转移以及转基因小鼠模型的评估。通过更充分了解增加的血管通透性的机制和该过程的逆转，我们将有能力开发针对AJ复合物组件的新型策略，以防止血管泄漏和肺水肿的形成。