Disintegrin Metalloprotease and Endothelial Permeability

描述（由申请人提供）：ADAM15是一种具有崩解素金属蛋白酶特征的跨膜糖蛋白，具有脱落受体分子和调节细胞-细胞/基质粘附的能力。ADAM15表达增加与转移性癌症、动脉粥样硬化和炎症性肠病的发生有关。我们最近的工作已经确定了这种分子的一种新功能，即能够增加血管内皮通透性的促炎因子。进一步的研究表明，在细菌感染或脓毒症刺激期间，ADAM15在肺部上调，导致中性粒细胞浸润和肺水肿。adam15介导的屏障损伤的组织特异性作用和潜在机制尚未明确。本项目的总体目标是了解ADAM15在炎症中的表达和功能的分子调控。我们提出了两个具体目标：1)表征ADAM15活性在脓毒症中的功能和调控途径；2)阐明ADAM15诱导高通透性的分子机制。需要验证的中心假设是，脓毒症在转录和翻译后水平诱导ADAM15上调。ADAM15通过胞质域激活Src信号转导增加内皮细胞-细胞连接通透性，而崩解素调控局灶粘连和/或连接结构的金属蛋白酶脱落是另一种途径。这一新颖的概念将通过一系列的补充研究得到验证，这些研究将利用动物模型进行体内生理分析，并在体外培养细胞中进行分子分析。将构建和测试创新的实验模型和分子工具。这项工作的数据不仅将建立蛋白酶分子生物学的新理论，而且还将为败血症和急性炎症的病理生理学提供新的机制见解。此外，该研究对识别和开发有效治疗炎症性疾病的新治疗靶点具有潜在影响。

DESCRIPTION (provided by applicant): ADAM15 is a transmembrane glycoprotein characteristic of disintegrin metalloprotease with the ability to shed receptor molecules and regulate cell-cell/matrix adhesions. Increased ADAM15 expression is associated with the development of metastatic cancer, atherosclerosis, and inflammatory bowl disease. Our recent work has identified a novel function of this molecule as a pro-inflammatory factor capable of increasing vascular endothelial permeability. Further studies suggest that ADAM15 is upregulated in the lungs during bacterial infection or septic stimulation, contributing to neutrophil infiltration and pulmonary edema. The tissue-specific effect and underlying mechanisms of ADAM15-mediated barrier injury have not been characterized. The overall goal of this project is to understand the molecular control of ADAM15 expression and function in inflammation. Two specific aims are proposed: 1) to characterize the functions and regulatory pathways of ADAM15 activity in sepsis, and 2) to elucidate the molecular mechanisms by which ADAM15 induces hyperpermeability. The central hypothesis to be tested is that sepsis induces ADAM15 upregulation at the transcriptional and posttranslational levels. ADAM15 increases endothelial cell-cell junction permeability by activating Src signal transduction via its cytoplasmic domain, whereas disintegrin regulation of focal adhesions and/or metalloprotease shedding of junctional structures serve as alternative pathways. This novel concept will be validated through a series of complementary studies that integrate in vivo physiological analyses using animal models and in vitro molecular assays in cultured cells. Innovative experimental models and molecular tools will be constructed and tested. Data derived from the proposed work will not only establish a novel theory in protease molecular biology, but also provide new mechanistic insights into the pathophysiology of sepsis and acute inflammation. In addition, the study has potential impact on the identification and development of novel therapeutic targets for effective treatment of inflammatory disease.