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、炎症和传染病。尽管大规模的努力提供了关于正常和疾病组织中许多不同蛋白酶表达的有价值的信息，但这些方法并没有报道蛋白酶的功能活性。蛋白酶信标正在成为一种非侵入性测量体内蛋白酶活性的强大工具，并且将受益于能够通过单个靶向蛋白酶进行更多选择性切割和探针保留在激活位点的方法。本项目旨在验证一种通用的分子探针设计，用于检测、定位和量化正常、病理组织和整个生物体体内的蛋白酶活性。具体来说，一种新的蛋白酶活性探针将被创造出来，它具有高组织摄取、个体靶蛋白酶选择性激活和激活位点积累的理想特征。将在正常小鼠和ApoE（-/-）小鼠动脉粥样硬化模型中证明这些探针在体内蛋白酶活性定位和定量方面的实用性。该项目的完成将产生准确可靠地报告参与正常和病理过程的蛋白酶活性的探针，具有很高的转化为人类的潜力。更广泛地说，成功完成本提案的目标将为具有高空间和时间精度的生物体中广泛的蛋白酶活性的测量和定位打开大门。

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. PUBLIC HEALTH RELEVANCE: This project aims to enable the detection, localization and quantification of a large class of enzymes that play key roles in disease processes, to enable completely new therapeutic approaches and diagnostic tools. We will develop and validate molecular probes that enable measurement of two protease activities that play crucial roles in the progression of cardiovascular disease. !