Measurement of protease activity in vivo

描述(由申请人提供)：已知蛋白酶活性在多种疾病的发病机制中起关键作用，包括癌症、心血管疾病、炎症性和感染性疾病。虽然大规模的研究提供了许多不同的蛋白水解酶在正常组织和疾病组织中表达的有价值的信息，但这些方法并没有报告蛋白水解酶的功能活性。蛋白水解酶信标正在成为一种非侵入性测量体内蛋白水解酶活性的强大工具，并将受益于能够通过个别靶向的蛋白水解酶进行更多选择性切割和在激活部位保留探针的方法。该项目旨在验证一种通用分子探针设计，以检测、定位和量化体内正常和病理组织以及整个生物体中的蛋白酶活性。具体地说，一类新的蛋白酶活性探针将被创建，它显示出所需的高组织摄取率、单个目标蛋白酶的选择性激活以及在激活部位积累的特征。在正常小鼠和动脉粥样硬化的载脂蛋白E(-/-)小鼠模型中，将证明这些探针用于定位和定量体内的蛋白酶活性。该项目的完成将产生准确和可靠地报告正常和病理过程中所涉及的蛋白酶活性的探针，具有很高的翻译到人类的潜力。更广泛地说，这项提议的目标的成功完成将为以高空间和时间精度测量和定位生物体中广泛的蛋白酶活性打开大门。

DESCRIPTION (provided by applicant): Protease activities are known to be critical in the pathogenesis of a wide variety of diseases including cancer, cardiovascular disease, and inflammatory and infectious diseases. Although large-scale efforts are providing valuable information regarding the expression of many diverse proteases in normal and disease tissues, these methods do not report the functional activity of proteases. Protease beacons are emerging as a powerful tool to non-invasively measure protease activity in vivo, and will benefit from approaches that enable more selective cleavage by individual targeted proteases and probe retention at the site of activation. This project aims to validate a generic molecular probe design to detect, localize, and quantify protease activities in vivo in normal and pathologic tissues and whole organisms. Specifically, a new class of protease activity probes will be created that exhibit the desired characteristics of high tissue uptake, selective activation by individual target proteases, and accumulation at sites of activation. The utility of these probes for the localization and quantification of protease activities in vivo will be demonstrated in normal mice and the ApoE(-/-) mouse model of atherosclerosis. Completion of this project will yield probes that accurately and reliably report the activity of proteases involved in normal and pathologic processes, with high potential for translational to humans. More generally, successful completion of the aims of this proposal will open the door to the measurement and localization of a wide range of protease activities in living organisms with high spatial and temporal precision.