Identifying the Molecular Mechanisms of Air Pollution-Induced Thrombosis

确定空气污染诱发血栓形成的分子机制

• 批准号: 10196986
• 负责人: Eva Vitucci
• 金额: $ 3.79万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10196986
• 关键词: ATF2 gene; Adverse effects; Air; Air Pollution; Alveolar; Anticoagulants; Antioxidants; Automobile Driving; Binding; Biological Assay; Biosensor; Blood Coagulation Factor; Blood capillaries; Cardiovascular Diseases; Cardiovascular system; Cessation of life; Chromatin; Coagulation Process; Coronary heart disease; Cysteine; Data; Deposition; Development; Diesel Exhaust; Distal; Endothelial Cells; Endothelium; Epithelial; Epithelial Cells; Exposure to; FOS gene; Fibrin; Fibrinolysis; Fibroblasts; Functional disorder; Future; Gene Expression; Generations; Genes; Genetic; Glutathione; Goals; Heart Rate; Homeostasis; Hydrogen Peroxide; Impairment; In Vitro; Intervention Studies; JUN gene; Kinetics; Link; Lung; MAP Kinase Gene; MAPK Signaling Pathway Pathway; MAPK8 gene; Mediating; Mediator of activation protein; Modeling; Molecular; Oxidation-Reduction; Oxidative Stress; Particulate; Particulate Matter; Pathology; Pathway interactions; Phenotype; Platelet Activation; Play; Populations at Risk; Prevention strategy; Principal Investigator; Production; Promoter Regions; Proteins; RNA; Role; Shapes; Signal Pathway; Signal Transduction; Stroke; Testing; Therapeutic; Thrombin; Thrombosis; Time; Training; Vascular Endothelial Cell; Vascular Endothelium; Work; activating transcription factor; alveolar epithelium; catalase-polyethylene glycol; cell type; chromatin immunoprecipitation; in vitro Model; in vivo; mortality; novel; oxidation; p38 Mitogen Activated Protein Kinase; p65; prevent; promoter; systemic inflammatory response; thrombotic; transcription factor; treatment strategy; vascular bed

Project Summary: Particulate matter (PM) air pollution is a common promoter for the development of thrombosis and cardiovascular disease. Specifically, exposure and deposition of PM in the alveolar capillary region (ACR) of the lung promotes thrombosis through platelet activation, impaired fibrinolysis, and increased coagulation factor production; however, the molecular mechanisms producing these adverse effects following PM exposure remain unclear. Our goal is to identify the vascular endothelium's role in promoting these thrombotic effects and elucidate the molecular mechanisms that result in a pro-thrombotic endothelium. To do this we have developed an organotypic in vitro model of the ACR in which we expose alveolar epithelial cells within the model to PM and determine the trans-epithelial (TE) effect in the underlying endothelial cells within the model. Our preliminary data suggests this TE exposure induces endothelial redox dysfunction, decreases the expression of anti-coagulant and fibrinolytic genes, and increases the expression of procoagulant genes. Our central hypothesis is TE exposure-induced redox dysfunction activates a pro-thrombotic endothelium by alternatively regulating the NF-κB and MAPK pathways. To test this hypothesis, our first aim is to define the relationship between the induced endothelial redox dysfunction and the activation of a pro-thrombotic endothelium. To define this relationship, we will compare the kinetics of redox dysfunction with the temporal expression of pro-thrombotic targets identified in our preliminary data and with the kinetics of functional thrombotic endpoints. Antioxidant intervention studies will then be performed to determine whether TE exposure-induced redox dysfunction drives the activation of a pro- thrombotic endothelium. Our second aim is to determine how the NF-κB and MAPK signaling pathways activate a thrombotic endothelium following a TE exposure. To determine these pathways' role we will evaluate the activation of the NF-κB, c-Jun N-terminal kinase (JNK), and p38 signaling pathways and will assess promoter binding at the activated pro-thrombotic targets of the downstream transcription factors of these pathways, p65, c-Fos/c-Jun, and ATF-2. Antioxidant intervention studies will then be performed to determine if TE-induced redox dysfunction mediates the activation of a pro-thrombotic endothelium by alternatively regulating these cellular pathways. The molecular mechanisms identified upon completion of these aims will help elucidate the vascular endothelium's role in promoting PMI-thrombosis. With these results, preventative and therapeutic strategies against PMI-thrombosis and related cardiovascular diseases can be developed. Completion of these aims will advance my training as a molecular toxicologist and support my development into a successful principal investigator.

项目概要： 颗粒物（PM）空气污染是血栓形成和心血管疾病发展的共同促进因素 疾病具体地，PM在肺的肺泡毛细血管区（ACR）中的暴露和沉积促进了肺的呼吸。 通过血小板活化、纤维蛋白溶解受损和凝血因子产生增加的血栓形成; 然而，PM暴露后产生这些不利影响的分子机制仍不清楚。 我们的目标是确定血管内皮在促进这些血栓形成作用中的作用，并阐明血管内皮在血栓形成中的作用。 导致促血栓形成内皮的分子机制。为了做到这一点，我们开发了一种器官型 ACR的体外模型，其中我们将模型内的肺泡上皮细胞暴露于PM，并测定 在该模型中，在底层内皮细胞中的跨上皮（TE）效应。我们的初步数据表明， TE暴露可诱导内皮细胞氧化还原功能障碍，降低抗凝和纤溶蛋白的表达， 基因，并增加促凝血基因的表达。我们的中心假设是TE干扰诱导的 氧化还原功能障碍通过交替调节NF-κB和MAPK激活血栓前内皮细胞 路径。为了验证这一假设，我们的第一个目标是确定诱导的内皮细胞之间的关系， 氧化还原功能障碍和促血栓形成内皮的激活。为了定义这种关系，我们将比较 氧化还原功能障碍的动力学与我们的研究中鉴定的促血栓形成靶点的时间表达有关， 初步数据和功能性血栓终点的动力学。抗氧化剂干预研究将 然后进行，以确定是否TE电刺激诱导的氧化还原功能障碍驱动激活的亲， 血栓形成内皮。我们的第二个目标是确定NF-κB和MAPK信号通路如何激活 TE暴露后血栓形成的内皮。为了确定这些途径的作用，我们将评估 NF-κB、c-Jun N-末端激酶（JNK）和p38信号通路的激活，并将评估启动子 结合在这些途径的下游转录因子的活化的促血栓形成靶点，p65， c-Fos/c-Jun和ATF-2。然后将进行抗氧化剂干预研究，以确定TE诱导的氧化还原是否 功能障碍通过交替调节这些细胞因子介导血栓形成前内皮的活化， 路径。完成这些目标后确定的分子机制将有助于阐明血管 内皮在促进PMI血栓形成中的作用。有了这些结果，预防和治疗策略 可以开发出抗PMI血栓形成和相关心血管疾病的药物。实现这些目标将 推进我作为分子毒理学家的培训，并支持我成为一名成功的校长 调查员