Structure-function of cytoprotective coagulation proteases and their receptors

项目摘要/摘要 这项应用旨在提高对介导细胞保护的分子机制的基本理解 凝血酶对细胞的作用。可用于治疗血管、血栓和 炎症性疾病是有限的，死亡率仍然高得令人无法接受。激活的有益效果 蛋白C(APC)对细胞的作用与其他凝血酶(如凝血酶)的促炎作用不同 和因子Xa)，并开创了对受体介导的复杂网络的新视角 细胞内的串扰。提高治疗的成功率需要对分子更彻底的了解。 涉及的机制。这一建议的核心是内皮蛋白C受体(EPCR)，这是一种关键 细胞保护性受体。此应用程序的长期目标是帮助诊断和 血管、血栓和炎症性疾病的治疗进展 包括基础研究和翻译研究。这个应用程序的目标是获得对分子的新见解 EPCR介导的细胞保护作用机制。此应用程序的主要焦点是结构- EPCRs的功能关系是EPCRs在临床相关基因转导中的辅因子作用的基础 APC介导的细胞保护作用及其转化为改进的治疗策略 用于血栓性炎症性疾病。新的假设将通过生物化学和细胞生物学进行检验。 方法：研究方法。具体目标是：1)产生具有独特性质的工程EPCR变体， 增强其介导细胞保护作用的能力，2)表征其结构-功能决定因素 依赖于EPCR的PAR-1激活，负责APC介导的细胞保护效应，以及 3)明确与“非靶标”肝素诱导的血小板减少症相关的血栓并发症 抗PF4-EPCR和PF4-血栓调节蛋白(TM)复合体的(HIT)抗体。圆满完成 拟议的研究将增加我们对血管、血栓和炎症的知识和理解 并可为开发针对各种疾病的新的治疗策略提供平台 血栓形成、细胞凋亡和炎症导致的疾病。

Project Summary/Abstract This application seeks to improve basic understanding of molecular mechanisms that mediate cytoprotective actions of coagulation proteases on cells. Available therapeutic solutions for vascular, thrombotic, and inflammatory diseases are limited and mortality rates remain unacceptably high. Beneficial effects of activated protein C (APC) on cells contrast with proinflammatory effects of other coagulation proteases (e.g. thrombin and factor Xa) on cells and initiated novel perspectives on the intricate complex networks of receptor-mediated cross talk in cells. Improving therapeutic success requires a more thorough understanding of the molecular mechanisms involved. This proposal is centered on the endothelial protein C receptor (EPCR), a key cytoprotective receptor. The long-term objectives of this application are to contribute to diagnostic and therapeutic progress for vascular, thrombotic, and inflammatory diseases by advancing knowledge through both basic and translational research. The goal of this application is to gain novel insights into the molecular mechanisms of cytoprotective actions mediated by EPCR. The major focus of this application is on structure- function relationships for EPCR that underlie EPCR's cofactor role in the transduction of clinically relevant APC-mediated cytoprotective effects and on translation of this information into improved therapeutic strategies for thrombotic inflammatory diseases. Novel hypotheses will be tested using biochemical and cellular biology methods. The specific aims are: 1) To generate engineered EPCR variants with unique properties that will enhance its ability to mediate cytoprotective effects, 2) To characterize the structure-function determinants for EPCR-dependent PAR-1 activation that are responsible for APC-mediated cytoprotective effects on cells, and 3) To define the thrombotic complications associated with "off-target" Heparin-Induced Thrombocytopenia (HIT) antibodies against PF4-EPCR and PF4-thrombomodulin (TM) complexes. Succesfull completion of the proposed studies will increase our knowledge and understanding of vascular, thrombotic, and inflammatory diseases and may provide a platform for the development of novel therapeutic strategies for a variety of disorders in which thrombosis, apoptosis and inflammation contribute to pathogenesis.