Measurement of protease activity in vivo

体内蛋白酶活性的测量

基本信息

批准号：
8238730
负责人：
Patrick S Daugherty
金额：
$ 41.29万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA BARBARA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-02-15 至 2016-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8238730
关键词：
Active Sites Address Affinity Animals Antibodies Antibody Binding Sites Apolipoprotein E Atherosclerosis Binding Biological Process Blocking Antibodies Cardiovascular Diseases Characteristics Communicable Diseases Data Detection Development Diagnostic Diagnostic Imaging Disease Engineering Enzymes Event Exhibits Family Fluorescence Resonance Energy Transfer Generic Drugs Half-Life Health Homologous Gene Hour Human Image Immunofluorescence Immunologic Immunoglobulin G In Vitro Individual Inflammatory Life Ligands MMP14 gene Magnetic Resonance Imaging Malignant Neoplasms Masks Measurement Measures Mediating Methods Molecular Probes Mus Organism Pathogenesis Pathologic Pathologic Processes Peptide Hydrolases Peptide Library Peptide antibodies Peptides Performance Physiological Physiological Processes Play Positron-Emission Tomography Process Regulation Reporting Role Site Specificity Thrombin Tissues Translating Whole Organism Yeasts base design directed evolution flexibility fluorescence imaging in vivo mouse model novel novel strategies novel therapeutic intervention polymerization scaffold tool uptake

项目摘要

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. !

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至关重要，包括癌症、心血管疾病以及炎症和传染病。尽管大规模的努力提供了有关正常和疾病组织中许多不同蛋白酶表达的有价值的信息，但这些方法没有报告蛋白酶的功能活性。蛋白酶信标正在成为非侵入性测量体内蛋白酶活性的强大工具，并将受益于能够通过单个目标蛋白酶进行更有选择性的切割以及探针保留在激活位点的方法。该项目旨在验证通用分子探针设计，以检测、定位和量化正常和病理组织以及整个生物体内的蛋白酶活性。具体来说，将创建一类新的蛋白酶活性探针，其表现出高组织摄取、单个靶蛋白酶选择性激活以及在激活位点积累的所需特征。这些探针在体内蛋白酶活性定位和定量方面的实用性将在正常小鼠和动脉粥样硬化 ApoE(-/-) 小鼠模型中得到证实。该项目的完成将产生能够准确可靠地报告正常和病理过程中涉及的蛋白酶活性的探针，并且具有向人类转化的巨大潜力。更一般地说，成功完成该提案的目标将为以高空间和时间精度测量和定位活生物体中的各种蛋白酶活性打开大门。公共健康相关性：该项目旨在实现对在疾病过程中发挥关键作用的一大类酶的检测、定位和定量，以实现全新的治疗方法和诊断工具。我们将开发并验证分子探针，以测量在心血管疾病进展中发挥关键作用的两种蛋白酶活性。！