EXTRACELLULAR PROTEASES IN INFLAMMATORY AIRWAY REMODELING

项目P-2的总体目标是检验细胞外蛋白酶是气道上皮腺体、细胞外基质和脉管系统重塑的关键决定因素这一假设，这些因素导致慢性气道炎症的持续存在和发病率。 Caughey博士和他的同事们将通过在转基因小鼠中对选定的蛋白酶及其靶点进行体内研究来解决这个问题，包括感染肺支原体作为慢性气道疾病模型的小鼠，以及对这些蛋白酶的分子行为和靶点进行体外研究。目的1：研究蛋白酶激活受体（PARs）在气道重塑中的作用，重点探讨凝血酶和肥大细胞类胰蛋白酶作为PAR-1和PAR-2的激活配体，可能介导这些蛋白酶对气道腺体和平滑肌的促生长作用。 这些受体在气道重塑中的作用将通过PAR表达在正常和发炎气道中的定位位点和探索PAR缺失小鼠中的气道重塑来检查。 目的2探讨蛋白酶在PAR基因敲除小鼠细胞外基质重塑中的作用。 目的2是探讨蛋白酶在慢性气道炎症细胞外基质重塑中的作用，特别是了解基质金属蛋白酶明胶酶B和巯基蛋白酶二肽基肽酶I的重要性。该实验室先前的实验表明，这两种蛋白酶都是由肥大细胞分泌和激活的，肥大细胞可能是这些酶在气道重塑中的重要来源。 在这些实验中，我们将定位蛋白酶在气道微环境中的表达，并研究支原体诱导的明胶酶和二肽基肽酶缺失小鼠的重塑。 目的3探讨肥大细胞蛋白酶在气道炎症时血管重塑中的作用，重点是蛋白酶介导的血管生成和血管抑制蛋白的形成和降解。 这里的方法是建立血管生成因子的分子机制和缺陷小鼠。 了解这些变化的机制可能会发现以前未探索的策略，以防止或逆转伴随慢性气道炎症的解剖学变化。

The overall goal of Project P-2 is to test the hypothesis that extracellular proteases are key determinants of the remodeling of the airway epithelium glands, extracellular matrix, and vasculature that contribute to the perpetuation of and morbidity from chronic airway inflammation. Dr. Caughey and his colleagues will address this issue by performing in vivo studies of selected proteases and their targets in genetically altered mice, including those infected with Mycoplasma pulmonis as a model of chronic airway disease, and in vitro studies of molecular behavior and targets of these proteases. Aim 1 is to determine the roles of proteinase-activated receptors (PARs) in airway remodeling, with a focus on thrombin and mast cell tryptase as activating ligands of PAR-1 and PAR-2, which may mediate growth-promoting effects of these proteases on airway glands and smooth muscle. The role of these receptors in airway remodeling will be examined by localization sites of PAR expression in normal and inflamed airways and exploring airway remodeling in PAR-null mice. Aim 2 is to explore roles of proteases in extracellular matrix remodeling in PAR-null mice. Aim 2 is to explore roles of proteases in extracellular matrix remodeling in chronic airway inflammation, seeking particularly to understand the importance of the matrix metalloproteinase gelatinase B and a thiol protease, dipeptidyl peptidase I. The laboratory's previous experiments suggest that both proteases are secreted and activated by mast cells, which may be an important source of these enzymes in airway remodeling. In these experiments we will localize proteases expression in airway microenvironments and examine mycoplasma-induced remodeling in gelatinase and dipeptidyl peptidase-null mice. Aim 3 is to explore the roles of mast cell proteases in vascular remodeling in airway inflammation, focusing on protease-mediated formation and degradation of angiogenic and angiostatic proteins. The approach here is to establish molecular mechanisms of angiotropic factor generation and to deficient mice. Understanding mechanisms underlying these changes may identify previously unexplored strategies to prevent or reverse anatomical changes accompanying chronic airway inflammation.