EXTRACELLULAR PROTEASES IN INFLAMMATORY AIRWAY REMODELING

项目P-2的总体目标是检验这一假设，即细胞外蛋白酶是呼吸道上皮腺、细胞外基质和血管重塑的关键决定因素，这些重塑有助于慢性呼吸道炎症的持久存在和发病率。考伊博士和他的同事们将通过在体内研究特定的蛋白水解酶及其靶点来解决这个问题，在基因改变的小鼠中，包括那些感染了肺支原体作为慢性呼吸道疾病模型的小鼠，以及对这些蛋白水解酶的分子行为和靶点进行体外研究。目的1探讨蛋白水解酶激活受体(PARs)在气道重塑中的作用，重点是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，它们可能介导这些酶对气道腺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在正常和炎症气道中的定位表达以及在PAR缺失小鼠中探索气道重塑来检验。目的2探讨蛋白水解酶在PAR缺失小鼠细胞外基质重塑中的作用。目的2探讨蛋白水解酶在慢性呼吸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和硫醇蛋白水解酶I的重要性。实验室先前的实验表明，这两种酶都是由肥大细胞分泌和激活的，这可能是这些酶在气道重塑中的一个重要来源。在这些实验中，我们将定位蛋白水解酶在呼吸道微环境中的表达，并检测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探讨肥大细胞蛋白水解酶在气道炎症血管重塑中的作用，重点探讨其在血管生成蛋白和血管抑制蛋白的形成和降解中的作用。这里的方法是建立血管生成因子和缺陷小鼠的分子机制。了解这些变化的潜在机制可能会确定以前未探索的预防或逆转伴随慢性呼吸道炎症的解剖变化的策略。

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.