Anti-Coagulant and Cytoprotective activity in CCM pathogenesis

CCM 发病机制中的抗凝血和细胞保护活性

基本信息

批准号：
10220146
负责人：
Mark HOWARD Ginsberg
金额：
$ 31.52万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10220146
关键词：
Acute Adult Anticoagulants Blocking Antibodies Blood Vessels Blood coagulation Brain CCM1 gene Chronic Deposition Development Disease Disease Progression Endothelial Cells Endothelium F2R gene Factor V Leiden mutation Hemophilia A Hemorrhage Hemosiderin Human Hypoxia Hypoxia Inducible Factor Impairment Intercellular Junctions Intervention Lead Lesion MSH2 gene Modeling Monoclonal Antibodies Morbidity - disease rate Mus Pathogenesis Pathway interactions Perinatal Pharmacology Preclinical Testing Protein C Research Resistance Role Sepsis Signal Transduction Source Stroke Symptoms Testing Thrombomodulin activated Protein C activated protein C receptor cerebral cavernous malformations factor V Leiden gene function insight loss of function mortality mutant neonatal mice neurovascular new therapeutic target prevent therapeutic target

项目摘要

ABSTRACT Cerebral Cavernous Malformations (CCM) are subject to acute and chronic bleeding that is a major source of morbidity and mortality in this disease. We have found that endothelial cells within murine and human CCM express markedly increased levels of thrombomodulin (TM) and endothelial protein C receptor (EPCR), which lead to activation of endogenous anti-coagulant protein C. We hypothesize that CCM form an anti-coagulant vascular domain and that activated Protein C (APC) contributes to bleeding in CCM. APC can also exert a cytoprotective effect on endothelium by signaling via PAR1 resulting in, among other effects, stabilization of endothelial cell-cell junctions. Indeed, this cytoprotective effect of APC has been exploited by creation of APC loss of function mutants that selectively maintain cytoprotective activity. We thus hypothesize that APC cytoprotective activity may limit morbidity from CCM as it does in experimental stroke. To genetically test this hypothesis, we will examine the bleeding in the acute murine CCM models in Factor V Leiden (F5R504Q/wt) mice, which are resistant to the anti-coagulant effect of APC. As a second approach we will test the effect of MAPC1591, a monoclonal antibody that blocks the anti-coagulant activity of APC in acute models of CCM. To test the effects of cytoprotective activity, we will examine acute CCM lesion development and bleeding in F2rR46Q/R46Q mice bearing PAR1 that is selectively resistant to APC cleavage. Conversely, we will examine the effect of 3K3A-APC, a loss of function APC mutant that is selectively impaired in anti-coagulant function in acute and chronic CCM models. We will exploit our observation that a brief period of hypoxia or pharmacological stabilization of Hypoxia-inducible factor 1A markedly exacerbates acute CCM formation in perinatal mice and enables a robust subacute model that manifests in adult mice. This subacute model which manifests both bleeding and hemosiderin deposition, will be used to test the interventions described in aims 1 and 2. Completion of these Aims will provide mechanistic insight into the role of the marked increase of TM and EPCR that is observed in CCM and provide important preclinical tests of the idea that hemorrhage or disease progression can be influenced by manipulating pathways that are already being therapeutically targeted in diseases such as stroke, sepsis, and hemophilia.

抽象的脑海绵状畸形（CCM）受到急性和慢性出血的约束该疾病中发病和死亡率的主要来源。我们发现内皮细胞在鼠和人类CCM中，明显增加了血栓形成蛋白水平（TM）和内皮蛋白C受体（EPCR），导致内源性抗凝蛋白的激活蛋白质C.我们假设CCM形成抗凝血剂血管结构域，并激活蛋白C（APC）有助于CCM出血。 APC也可以对通过PAR1发出信号的内皮，除其他效果外，内皮稳定细胞电池连接。实际上，APC的这种细胞保护作用已通过创建来利用 APC的功能突变体丧失，有选择地维持细胞保护活性。我们这样假设APC细胞保护活性可能会限制CCM的发病率实验性中风。为了基因检验这一假设，我们将检查急性的出血因子V Leiden（F5R504Q/WT）小鼠中的鼠CCM模型，对抗凝凝剂具有抗药性 APC的效果。作为第二种方法，我们将测试MAPC1591（单克隆抗体）的效果这阻断了CCM急性模型中APC的抗凝活性。测试效果细胞保护活性，我们将检查急性CCM病变的发育和出血 F2RR46Q/R46Q携带PAR1的小鼠对APC裂解具有抗性。相反，我们会的检查3K3A-APC的效果，该功能损失APC突变体在选择性上受到损害急性和慢性CCM模型中的抗凝功能。我们将利用我们的观察缺氧或缺氧诱导因子1A的短暂短暂或药理稳定时期明显加剧围产期小鼠的急性CCM形成，并实现强大的亚急性模型在成年小鼠中表现出来。这种表现出出血和脱皮蛋白的亚急性模型沉积将用于测试目标1和2中描述的干预措施。目的将提供机械洞察力，了解TM和EPCR明显增加的作用在CCM中观察到，并提供了重要的临床前测试，即出血或疾病进展可能会受到已经正在治疗的操纵途径的影响针对中风，败血症和血友病等疾病。