Lipid peroxidation- and pyroptosis-induced tissue factor activation in pathogen-induced blood coagulation

病原体诱导的血液凝固中脂质过氧化和焦亡诱导的组织因子激活

基本信息

批准号：
10571353
负责人：
Shabbir Ahmed Ansari
金额：
$ 16.45万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10571353
关键词：
2019-nCoV 4 hydroxynonenal Acute myocardial infarction Adult Affect Aldehydes Apoptosis Arachidonate 15-Lipoxygenase Atherosclerosis Attenuated Bacteremia Bacterial Infections Blood coagulation CASP1 gene Caspase Cell Death Cell surface Cessation of life Coagulation Process Data Disease Disseminated Intravascular Coagulation Disulfiram Enzymes Epithelium Escherichia coli Evaluation FDA approved Functional disorder Generations Genes Host Defense Human In Vitro Infection Inflammasome Inflammatory Ischemic Stroke LOX gene Life Lipid Peroxidation Lung infections Macrophage Mediating Modeling Multiple Organ Failure Mus Myocardial Infarction Organ Pathogenicity Pathologic Pathway interactions Phase Phosphatidylserines Play Polyunsaturated Fatty Acids Positioning Attribute Process Proteins Pulmonary Embolism Reporting Resistance Role SARS-CoV-2 infection Sepsis Testing Therapeutically Targetable Thromboplastin Thrombosis Training United States Virus Diseases Work chemical genetics extracellular vesicles genetic approach human disease improved in vivo lipid metabolism monocyte mouse model new therapeutic target pathogen peroxidation prevent septic thrombotic vesicular release

项目摘要

Project Summary/Abstract Tissue factor (TF), the primary initiator of the blood coagulation cascade, is carefully regulated to prevent aberrant coagulation activation. However, pathological conditions including bacterial and viral infection induce intravascular TF expression and lead to thrombosis. As TF-induced thrombosis is a major cause of acute myocardial infarction, ischemic stroke, and pulmonary embolism, improved understanding of the mechanisms of pathological TF activation may lead to new therapeutic targets. Pyroptosis, a form of inflammatory cell death, drives TF-mediated intravascular coagulation activation in bacterial sepsis. Emerging studies also implicate inflammasome activation in SARS-CoV-2 infection, but its role in TF activation is unknown. Our preliminary studies demonstrate that SARS-CoV-2 and specifically its accessory protein ORF3A induce TF activation in a phosphatidylserine (PS)-dependent mechanism that requires TMEM16F, similar to PS-dependent TF activation in pyroptosis. In Aim 1, we will investigate whether ORF3a-induced TF activation is driven by inflammasome- mediated pyroptosis. Lipid peroxidation and its highly reactive end products such as 4-hydroxy-2-nonenal (HNE) are involved in various forms of programmed cell death including pyroptosis. However, the role of HNE, the most stable and toxic reactive aldehyde produced during lipid peroxidation, in pyroptosis-associated TF and coagulation activation is not known. Our preliminary data showed that HNE induces PS-dependent TF activation in LPS-primed macrophages and causes intravascular coagulation activation in mice. However, the complete mechanism by which HNE induces PS externalization and TF activation is not known. In Aim 2, we will use chemical genetic approaches to dissect the mechanism of HNE-induced TF activation via pyroptosis in vitro and in vivo. Although lipid peroxidation plays a central role in cell death and coagulation activation in bacterial sepsis, a therapeutically targetable enzyme responsible for the unbridled lipid peroxidation and generation of pathological levels of reactive radicals such as HNE is not known. In Aim 3, we will use genetically modified mice deficient in lipid peroxidation and HNE formation to investigate TF-dependent pathologic coagulation activation and thrombosis during sepsis. A successful completion of these studies will help delineate a common pathway involved in pathologic TF activation across varied pathogenic infections and will also help identify a specific therapeutically targetable enzyme to attenuate TF activation in disease.

项目摘要/摘要组织因子（TF）是血液凝结级联反应的主要启动因子，仔细调节以防止异常凝血激活。但是，包括细菌和病毒感染在内的病理状况会诱导血管内TF表达并导致血栓形成。因为TF诱导的血栓形成是急性的主要原因心肌梗塞，缺血性中风和肺栓塞，改善了对机制的理解病理TF激活可能导致新的治疗靶标。凋亡，一种炎症细胞死亡的形式，驱动TF介导的细菌败血症中的血管内凝血活化。新兴研究也暗示 SARS-COV-2感染中的炎性体激活，但其在TF激活中的作用尚不清楚。我们的初步研究表明，SARS-COV-2及其辅助蛋白ORF3A在A中诱导TF激活需要TMEM16F的磷脂酰丝氨酸（PS）依赖性机制，类似于PS依赖性TF激活在凋亡中。在AIM 1中，我们将调查ORF3A诱导的TF激活是否由炎性组驱动介导的凋亡。脂质过氧化及其高反应性最终产物，例如4-羟基-2-非纳尔（HNE）涉及各种形式的程序性细胞死亡，包括凋亡。但是，HNE的角色，最大脂质过氧化过程中产生的稳定和毒性的活性醛在与凋亡相关的TF和凝血激活尚不清楚。我们的初步数据表明，HNE诱导PS依赖性TF激活在LPS化力的巨噬细胞中，引起小鼠的血管内凝血活化。但是，完整 HNE诱导PS外部化和TF激活的机制尚不清楚。在AIM 2中，我们将使用化学遗传学方法是通过体外凋亡和体内。尽管脂质过氧化在细菌败血症的细胞死亡和凝结激活中起着核心作用，但一种具有治疗靶向的酶，负责无限制的脂质过氧化和产生反应性自由基（例如HNE）的病理水平尚不清楚。在AIM 3中，我们将使用转基因的小鼠缺乏脂质过氧化和HNE形成，以研究TF依赖性病理凝结激活和败血症期间的血栓形成。这些研究的成功完成将有助于描述一个通用的途径参与各种致病感染的病理TF激活，也将有助于确定特定具有治疗靶向酶可以减弱疾病中的TF激活。