Identifying the Molecular Mechanisms of Air Pollution-Induced Thrombosis

项目总结： 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剂 疾病。具体地说，PM暴露和沉积在肺泡毛细血管区域(ACR)促进 通过血小板活化、纤溶功能受损和凝血因子产生增加而形成血栓； 然而，PM暴露后产生这些不良影响的分子机制仍不清楚。 我们的目标是确定血管内皮细胞在促进血栓形成方面的作用，并阐明 导致血栓形成的内皮细胞的分子机制。为了做到这一点，我们开发了一种有机类型 在ACR的体外模型中，我们将模型中的肺泡上皮细胞暴露于PM，并确定 跨上皮(TE)效应在模型内的潜在内皮细胞。我们的初步数据表明 TE暴露致内皮细胞氧化还原功能障碍，抗凝、纤溶活性降低 基因，并增加促凝血基因的表达。我们的中心假设是TE暴露诱导的 氧化还原功能障碍通过交替调节NF-κB和MAPK激活血栓形成前血管内皮细胞 小路。为了验证这一假设，我们的第一个目标是定义诱导的内皮细胞与 氧化还原功能障碍和促血栓内皮细胞的激活。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促血栓形成靶点在OUR中的时间表达 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 然后进行检查，以确定TE暴露诱导的氧化还原功能障碍是否驱动了一种前- 血栓内皮细胞。我们的第二个目标是确定NF-κB和MAPK信号通路是如何激活的 血栓内皮细胞在血栓栓塞术后暴露。为了确定这些途径的作用，我们将评估 激活核因子-κB、c-jun氨基末端激酶和p38信号通路，并将评估启动子 结合在这些途径下游转录因子的激活的促血栓形成靶点，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 功能障碍通过交替调节这些细胞来介导血栓前内皮细胞的激活 小路。完成这些目标后确定的分子机制将有助于阐明血管 内皮细胞在促进PMI-血栓形成中的作用。根据这些结果，预防和治疗策略 抗PMI-血栓形成和相关的心血管疾病可以开发出来。完成这些目标将 提升我作为分子毒物学家的培训，支持我成长为一名成功的校长 调查员。

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.