Structure-function of cytoprotective coagulation proteases and their receptors

项目概要/摘要 本申请旨在提高对介导细胞保护作用的分子机制的基本理解。 凝固蛋白酶对细胞的作用。现有的血管、血栓和 炎性疾病有限，死亡率仍然高得令人无法接受。激活的有益效果 蛋白C（APC）对细胞的促炎作用与其它凝血蛋白酶（例如凝血酶）的促炎作用形成对比 和因子Xa）对细胞的作用，并引发了对受体介导的复杂网络的新观点。 细胞内的串扰。提高治疗成功率需要更彻底地了解分子水平， 涉及的机制。该提案以内皮蛋白C受体（EPCR）为中心， 细胞保护受体本申请的长期目标是有助于诊断和 血管、血栓和炎性疾病的治疗进展， 基础研究和转化研究。本申请的目标是获得对分子生物学的新见解。 EPCR介导的细胞保护作用的机制。这个应用程序的主要重点是结构- EPCR的功能关系，EPCR的辅因子的作用，在转导临床相关的 APC介导的细胞保护作用以及将此信息转化为改进的治疗策略 用于血栓性炎症性疾病。新的假设将使用生物化学和细胞生物学进行测试 方法.具体目标是：1）产生具有独特性质的工程化EPCR变体， 增强其介导细胞保护作用的能力，2）表征结构-功能决定因素， EPCR依赖性PAR-1激活，负责APC介导的细胞保护作用，和 3)定义与“脱靶”肝素诱导的血小板减少症相关的血栓性并发症 (HIT)抗PF 4-EPCR和PF 4-血栓调节蛋白（TM）复合物的抗体。圆满完成了 拟议的研究将增加我们对血管、血栓和炎症的认识和理解， 并且可以为开发用于各种疾病的新治疗策略提供平台。 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.