Regulation of Protein C Pathways

蛋白 C 通路的调节

• 批准号: 10604355
• 负责人: JOHN H GRIFFIN
• 金额: $ 88.3万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10604355
• 关键词: 2019-nCoV; Abbreviations; Acids; Acute; Address; Affect; Age; Alteplase; Animals; Anticoagulants; Apoptosis; Biological; Biological Products; Blocking Antibodies; Blood Coagulation Disorders; Blood coagulation; COVID-19; COVID-19 complications; COVID-19 treatment; Carboxypeptidase U; Cardiovascular Diseases; Cells; Cessation of life; Chronic; Clinic; Coagulation Process; Collection; Cytoprotection; Data; Databases; Defense Mechanisms; Development; Diabetes Mellitus; Disease; Endothelium; Engineering; F2R gene; Factor V; Factor X; Feedback; Fibrinolysis; Future; Generations; Heart Injuries; Homeostasis; Host Defense; Hyperactivity; Hypertension; ITGAM gene; ITGB2 gene; Immune; Immune Cell Suppression; In Vitro; Inflammation; Inflammatory; Injury; Injury to Kidney; Integrins; Ischemia; Ischemic Stroke; Islets of Langerhans Transplantation; Kidney Transplantation; Knowledge; Laceration; Libraries; Life; Liver; Lung infections; Macrophage-1 Antigen; Mediating; Modeling; Molecular; Mus; Nervous System Trauma; Organ; Outcome; PAR-1 Receptor; Pathology; Pathway interactions; Peptide Hydrolases; Plasma; Plasminogen Activator Inhibitor 1; Play; Process; Protein C; Protein C Inhibitor; Protein S; Proteinase-Activated Receptors; Pseudomonas; Reaction; Reagent; Recombinants; Regulation; Reperfusion Injury; Role; SARS-CoV-2 infection; Sepsis; Serine Protease; Shock; Signal Induction; Signal Transduction; Specificity; Symptoms; System; Testing; Therapeutic; Thrombomodulin; Thrombophilia; Tissues; Translating; Translations; Trauma; Trypsin; Variant; Whole-Body Irradiation; activated Protein C; activated protein C receptor; cell injury; cell type; citrate carrier; clinical development; comorbidity; enzyme activity; gastrointestinal; graft vs host disease; immunothrombosis; improved; in vivo; insight; lung injury; man; mortality; mutant; novel; pharmacologic; pre-clinical; preclinical study; protein protein interaction; receptor; severe COVID-19; success; targeted treatment; thromboinflammation; tool; translational potential; trauma induced coagulopathy

Project Summary/Abstract Activated protein C (APC) is a naturally occurring plasma serine protease that has been translated to the clinic as a recombinant wild type or mutant biologic. In a diverse collection of preclinical animal injury models, pharmacologic APC’s provides many benefits. APC and the protein C systems contribute to the regulation of thromboinflammation that is critical for host defense, including defense versus SARS-CoV-2 infection. APC can act on diverse cell types, including immune cells for suppression of graft-versus-host disease. APC not only has anticoagulant activity but also initiates cell signaling via multiple receptors, in particular, via several protease activated receptors (PAR). APC-initiated cell signaling contributes to tissue homeostasis and host defense systems. Beneficial APC-initiated biased signaling is caused by specific cleavages of PAR1 and PAR3, and for some critical biologic activities, e.g., reducing mortality in sepsis, APC requires the integrin, Mac-1. Despite recent insights, there is a major gap in knowledge about protein-protein interactions between APC and its cellular receptors. Aim 1 studies will use a very extensive library of > five dozen recombinant APC mutants to provide a database regarding APC’s receptor specificities which will then enable engineering of APC mutants with receptor-specific selectivity, e.g., an APC mutant with highly selective PAR1-specific or PAR3-specific signaling capabilities or selectively specific for the I-domain of Mac-1. Such receptor-selective APC mutants will be useful reagents for deciphering which receptors play critical roles on cells in vitro or in animals in vivo, and they may lead to translation for novel APC mutants. To address the major problem of COVID-19, Aim 2 studies will characterize how murine COVID-19-like symptoms in SARS-CoV-2 infected hACE2 mice are influenced by endogenous protein C systems, e.g., by anticoagulant systems, by cytoprotective systems including anti- inflammation, anti-apoptosis, and endothelial barrier stabilization, and by fibrinolysis systems. Studies will define how these systems are modulated to be either hyperactive or hypoactive. This will provide new insights into how the protein C systems may be intertwined with defense versus SARS-CoV-2. The most promising cytoprotective- selective APC mutant (3K3A-APC) will be tested for COVID-19 therapy to advance potential translation. Aim 3 will characterize the multifaceted roles of APC in the pathobiology of acute traumatic coagulopathy. Studies will assess mechanisms by which disproportional APC generation contributes either to the development of acute traumatic coagulopathy involving Factor V inactivation and hyperfibrinolysis or, conversely, to protection of the integrity of organ function during and after acute traumatic coagulopathy by way of APC’s cytoprotective activities. The proposed studies will provide novel mechanistic insights and new APC variants which may aid translations to the clinic related to the APC pathways.

项目摘要/摘要 活化蛋白C(APC)是一种天然存在的血浆丝氨酸蛋白酶，已被翻译到临床上 作为重组野生型或突变型生物。在一系列临床前动物损伤模型中， 药理上的APC提供了许多好处。APC和蛋白C系统有助于调节 对宿主防御至关重要的血栓性炎症，包括对SARS-CoV-2感染的防御。APC可以 作用于不同类型的细胞，包括免疫细胞，以抑制移植物抗宿主疾病。APC不仅拥有 抗凝活性，但也通过多个受体，特别是通过几种蛋白水解酶来启动细胞信号 激活受体(PAR)。APC启动的细胞信号转导有助于组织动态平衡和宿主防御 系统。有益的APC启动的有偏见的信号是由PAR1和PAR3的特定切割引起的，并且对于 一些关键的生物学活动，如降低脓毒症的死亡率，APC需要整合素，Mac-1。尽管 最新的见解，在APC和其细胞之间的蛋白质-蛋白质相互作用的知识上存在着重大的差距 感受器。Aim 1的研究将使用一个非常广泛的&gt；56个重组APC突变体的文库来提供 有关APC受体特异性的数据库，这将使APC突变体的工程具有 受体特异性选择性，例如，具有高度选择性PAR1或PAR3特异性信号的APC突变体 能力或选择性地特定于MAC-1的i域。这种受体选择性的APC突变体将是有用的 用于破译在体外或在动物体内的细胞上起关键作用的受体的试剂，它们可能 导致新的APC突变体的翻译。为了解决新冠肺炎的主要问题，Aim 2研究将 描述SARS-CoV-2感染的hACE2小鼠的新冠肺炎样症状是如何受到 内源性蛋白C系统，例如通过抗凝剂系统，通过包括抗凝剂系统在内的细胞保护系统 炎症、抗细胞凋亡和稳定内皮屏障，以及纤溶系统。研究将定义 这些系统是如何被调节为过度活跃或低活的。这将提供关于如何 蛋白C系统可能与对抗SARS-CoV-2的防御系统交织在一起。最有希望的细胞保护- 选择性抗癌基因突变体(3K3A-抗癌基因)将接受新冠肺炎治疗的测试，以促进潜在的翻译。目标3 将描述APC在急性创伤性凝血病的病理生物学中的多方面作用。研究将会 评估不成比例的APC生成在急性胰腺炎发病中的作用机制 创伤性凝血病涉及凝血因子V失活和纤溶亢进，或者相反，与保护 APC细胞保护作用在急性创伤性凝血障碍中和术后器官功能的完整性 活动。拟议的研究将提供新的机理见解和新的APC变体，这可能有助于 翻译到与APC通路相关的诊所。