Association of tissue factor pathway inhibitor with endothelium

组织因子途径抑制剂与内皮细胞的关联

• 批准号: 8008763
• 负责人: Alan E Mast
• 金额: $ 36.68万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8008763
• 关键词: Adult; Adverse effects; Anti-Inflammatory Agents; Anti-inflammatory; Anticoagulants; Binding; Biological Assay; Biological Models; Blood Clot; Blood coagulation; Breeding; C-terminal; Caveolae; Cell surface; Cells; Chinese Hamster Ovary Cell; Coagulants; Complex; Development; Disease; Disseminated Intravascular Coagulation; Drug or chemical Tissue Distribution; Embryo; Endothelial Cells; Endothelium; Equilibrium; Factor IX; Factor VIIa; Glycosylphosphatidylinositols; Goals; Health; Hemorrhage; Human; In Vitro; Individual; Inflammation; Inflammatory; Knockout Mice; Mediating; Membrane; Methods; Mus; Myocardial Infarction; Neoplasm Metastasis; Pathogenesis; Pathology; Peptide Hydrolases; Physiological; Placenta; Prevention; Production; Protein Isoforms; Proteinase-Activated Receptors; Proteins; Public Health; RNA Splicing; Receptor Signaling; Resistance; Stroke; Structure; Surface; System; TFPI; Tetracycline Control; Thromboplastin; Thrombosis; Time; Tissues; Trans-Activators; Transgenic Mice; aerolysin; base; caveolin 1; design; human tissue; in vitro activity; in vivo; in vivo Model; inhibitor/antagonist; prevent; receptor; receptor binding; response to injury; tumor

DESCRIPTION (provided by applicant): Tissue Factor Pathway Inhibitor (TFPI) is an endothelial- associated anticoagulant protein that directly inhibits the Tissue Factor-factor VIIa (TF-fVIIa) catalytic complex preventing intravascular thrombosis, disseminated intravascular coagulation, tumor metastasis and other pathologies associated with production of TF within the vasculature. TF-fVIIa activates blood clotting via cleavage of factors IX and X and initiates inflammation via activation of Protease Activated Receptors (PARs). Our long-term goal is to develop methods for prevention and treatment of the adverse effects of intravascular TF activity based upon understanding its pathobiology through studies of its inhibition by TFPI. TFPI is made in three alternatively spliced isoforms that differ in their tissue distribution, domain structure, and mechanism for cell surface association. We hypothesize that examination of structurally diverse forms of TFPI with distinct mechanisms for cell surface association will reveal variable efficacies in their ability to inhibit TF-mediated pro-coagulant and/or pro-inflammatory activity. By characterizing the unique features of how TFPI down-regulates TF activity we will better understand the pathogenesis of diseases mediated by intravascular expression of TF. Three Specific Aims are designed to define the functions of different forms of TFPI using both in vitro and in vivo model systems. SA1 will compare the anti-TF activity of different forms of GPI-anchored TFPI on the surface of CHO cells and primary mouse endothelial cells. SA2 will examine how the mode of membrane association (GPI-anchored vs. transmembrane) TFPI activity on CHO cells, aerolysin resistant EA.hy926 cells and primary mouse endothelial cells. Studies will also be performed to examine how caveolin-1 and the GPI-anchored co- receptor for TFPI-alpha impact TFPI inhibitory activity. In SA3 we will produce transgenic mice specifically expressing TFPI-alpha or TFPI-beta using an inducible and reversible tetracycline controlled transactivator system. The transgenic mice will be bred with TFPI() mice to determine if there is isoform specific rescue of embryonic lethality. PUBLIC HEALTH RELEVANCE: This proposal is relevant to public health as it investigates cellular responses to injury and inflammation, balancing the development of blood clots necessary to prevent severe bleeding while at the same time avoiding the development of intravascular blood clots that can result in heart attack or stroke and enhance tumor metastasis.

描述（由申请人提供）：组织因子途径抑制剂（TFPI）是一种内皮 - 相关的抗凝剂蛋白，直接抑制组织因子因子因子VIIA（TF-FVIIA）催化复合物，可防止抗血管内血栓形成，可预防血管内血栓形成，与themoly pationlogies相关，themory pateragilies and themologies and themologies，tumor paterations Inter In In In In Intature in tff ins tff。 TF-FVIIA通过裂解IX和X的裂解激活血液，并通过激活蛋白酶激活受体（PARS）引发炎症。我们的长期目标是基于对TFPI抑制作用的理解，开发用于预防和治疗血管内TF活性的不良反应的方法。 TFPI是在三个剪接的同工型中制成的，它们的组织分布，结构域结构和细胞表面缔合机制不同。我们假设检查具有不同机制的结构多样性的TFPI，将揭示其抑制TF介导的促凝剂和/或促炎活性的能力。通过表征TFPI如何降低TF活性的独特特征，我们将更好地理解由TF的血管内表达介导的疾病的发病机理。设计了三个特定的目标，旨在使用体外和体内模型系统来定义不同形式的TFPI的功能。 SA1将比较不同形式的GPI锚定TFPI在CHO细胞表面和原代小鼠内皮细胞表面的抗TF活性。 SA2将检查膜关联模式（GPI锚定与跨膜）如何对CHO细胞，耐气蛋白EA.HY926细胞和原代小鼠内皮细胞的TFPI活性。还将进行研究以检查Caveolin-1和GPI锚定的TFPI-Alpha撞击TFPI抑制活性的受体。在SA3中，我们将使用可诱导和可逆的四环素控制的反式激活器系统来产生专门表达TFPI-Alpha或TFPI-beta的转基因小鼠。转基因小鼠将用TFPI（）小鼠繁殖，以确定是否存在同工型特异性拯救胚胎致死性。公共卫生相关性：该提案与公共卫生有关，因为它调查了细胞对损伤和炎症的反应，平衡了防止严重出血所需的血凝块的发展，同时避免了可能导致心脏病或中风的血管内血液凝块的发展，并增强了肿瘤转移。