Focal adhesion kinase regulation of lung vascular permeability and edemagenesis

描述（由申请人提供）：肺血管屏障功能丧失和炎性细胞浸润是急性肺损伤（ALI）的两个主要特征。本课题旨在探讨内皮细胞粘着斑激酶（endothelial focal adhesion kinase，FAK）通过抑制细胞内Ca ~（2+）升高来调节肺血管通透性增加和炎症反应的基本原理。我们将检验FAK诱导靶向Ca 2+传感器基质相互作用分子1（STIM 1）的microRNA-150（miR-150）表达的假设。我们推测，通过这种机制下调STIM 1对于阻断内皮细胞中的Ca 2+信号传导至关重要，从而防止肺血管屏障和炎症的破坏。为了解决这一假设，我们已经产生了一种新的他莫昔芬诱导的内皮细胞（EC）特异性FAK敲除（EC-FAK-/-）以及EC-STIM 1-/-小鼠模型。我们将致力于以下目的：目的#1，使用内皮细胞特异性FAK缺失小鼠来确定FAK在调节肺液体平衡和调节肺部炎症中的作用;目的#2，确定FAK在抑制STIM 1活性并由此阻断介导肺血管通透性增加和炎症的Ca 2+进入信号中的作用;和目标#3，确定miR-150作为FAK的重要效应物的作用，其靶向STIM 1，从而防止肺血管通透性增加和炎症。这些研究将使用成像，遗传和生理方法的组合，以开发一个全面和综合的图片内皮FAK在调节肺液稳态和炎症的作用。这些研究的结果将有可能确定针对ALI的新的治疗靶点。 公共卫生相关性：急性肺损伤（acute lung injury，ALI）是一种以血浆渗漏、白色血细胞内流增加和细胞因子生成增加为特征的危及生命的疾病。尽管在有效支持ALI患者方面取得了很大进展，但目前的治疗策略在预防ALI方面的成功有限。因此，继续寻找更有效的治疗方法，以降低由ALI引起的不可接受的高死亡率。拟开展的研究将探讨内皮蛋白、粘着斑激酶（FAK）在维持肺液稳态和减轻肺炎症损伤中的作用。我们希望这些研究将提供新的见解和治疗目标，我们认为这将导致开发更有效的治疗方法，改善ALI患者的预后。

DESCRIPTION (provided by applicant): Loss of lung vascular barrier function and inflammatory cell infiltration are two cardinal features of acute lung injury (ALI). The project wil investigate the fundamental concept that endothelial focal adhesion kinase (FAK) modulates the increase in lung vascular permeability and the inflammatory response by suppressing intracellular Ca2+ rise. We will test the hypothesis that FAK induces microRNA-150 (miR-150) expression which targets the Ca2+ sensor, stromal interacting molecule 1 (STIM1). We postulate that STIM1 downregulation by this mechanism is crucial for blocking Ca2+ signaling in endothelial cells and thereby prevents disruption of lung vascular barrier and inflammation. To address this hypothesis, we have generated a novel tamoxifen-inducible endothelial cell (EC)-specific FAK knockout (EC-FAK-/-) as well as EC-STIM1-/- mouse models. We will address the following aims: Aim #1, To define the role of FAK in regulating lung fluid balance and modulating lung inflammation using endothelial cell-specific FAK null mice; Aim #2, To determine the role of FAK in suppressing STIM1 activity and thereby blocking Ca2+ entry signals mediating increased lung vascular permeability and inflammation; and Aim #3, To define the role of miR-150 as an essential effector of FAK that targets STIM1 and thus prevents increased lung vascular permeability and inflammation. These studies will use a combination of imaging, genetic, and physiological approaches to develop a comprehensive and integrated picture of the role of endothelial FAK in regulating lung fluid homeostasis and inflammation. The results of these studies will have the potential of defining novel therapeutic targets directed against ALI. PUBLIC HEALTH RELEVANCE: Acute lung injury (ALI) characterized by plasma leakage, increased white blood cell influx and cytokine generation is a life threatening disease. Although great strides have been made in effectively supporting ALI patients, current treatment strategies have demonstrated limited success in preventing ALI. Therefore the search continues for more effective treatments to reduce the unacceptably high rate of mortality caused by ALI. Proposed studies will investigate the role of endothelial protein, focal adhesion kinase (FAK) in maintaining lung fluid homeostasis and moderating lung inflammatory injury. We hope these studies will offer novel insights and therapeutic targets that we think will lead to the developmen of more effective treatments with improved outcomes for patients suffering from ALI.