Project 2

项目2摘要 项目2是一个新的生物医学研究项目，旨在开发新的工具来快速表征儿科 接触有害挥发性有机化合物(VOCs)带来的呼吸道健康风险。这项工作将是一项 德州农工大学超级基金研究中心总体战略中的关键要素 并管理与暴露在环境应急动员的危险环境中相关的人类健康风险 物质。目前的毒性测试策略没有考虑到具有代表性的 尽管这些因素(即，年龄、性别、种族、 和遗传)是哮喘风险的关键因素。此外，单个/组合VOCs背后的作用机制 关于哮喘的发病机制目前还知之甚少。支持对危险VOCs的评估，包括REAL 城市混合物，并阐明机械联系，项目2将测试假设，儿科呼吸道是 对有害VOCs的肺损伤非常敏感，而且呼吸道反应是由 细胞外小泡(EV)介导的信号转导。研究小组汇集了一名毒物学家，一名内科医生- 一名科学家和一名大气化学家，以解决以下具体目标。目标1优先评估 20个超级基金优先VOCs在体外测试哮喘相关表型，首先使用呼吸系统 在气液界面培养上皮细胞系(16HBE)，然后在基于群体的适龄模型中培养 由发展中的肺分子图谱计划中的儿科支气管上皮细胞组成。下一首, 与环境空气中与环境相关的化学物质比例相匹配的代表性设计混合物将 在标准的和基于群体的儿科细胞系中进行评估。这些回应将告知AIM 2 机制研究，将测试VOC暴露改变EV蛋白表达的假设，潜在的 呼吸功能障碍。已知炎症和上皮屏障功能是由外切体介导的， 一类从30 nm到150 nm的分泌型电动汽车。在AIM 2中，来自16HBE细胞的EV暴露于选择 将使用高通量蛋白质组学方法对VOCs/混合物进行纯化和测序。蛋白质签名 这个模型中揭示的结果将在不同的儿科捐赠者细胞系中得到验证。最后，政府部门的职能作用 暴露在VOC中的细胞衍生电动汽车将通过过继转移实验进行评估。与AIMS 1和AIMS并行 2、Aim 3目标将通过在不同地点的移动空气监测来表征VOC混合物 在基线期间和在应对环境灾难时，扩大休斯顿地区。此外，为了填补空白， 灾难相关毒性测试，16HBE电池将直接暴露在移动平台上的环境空气中 在野外，使用时间分辨测量来驱动对不同气团的条件采样。结果 项目2中提出的研究与超级基金计划高度相关。总体而言，新的工具和 这些发现将增进对VOC所致儿童肺损伤机制的基本了解 并使改进的风险评估能够迅速确定威胁儿童健康的呼吸道危害的特征。

Project 2 ABSTRACT Project 2 is a new Biomedical Research Project aimed at developing novel tools to rapidly characterize pediatric respiratory health risks from exposure to hazardous volatile organic compounds (VOCs). This work will be a critical element in the overall strategy of the Texas A&M University Superfund Research Center to characterize and manage the human health risks associated with exposure to environmental emergency-mobilized hazardous substances. Current toxicity testing strategies do not account for developmental stage that is representative of the pediatric lung or encompass human population variability, even though these factors (i.e., age, sex, race, and genetics) are critical in asthma risk. Moreover, mechanisms of action underlying individual/combined VOCs on asthma pathogenesis is poorly understood. To support the evaluation of hazardous VOCs, including real urban mixtures, and elucidate mechanistic linkages, Project 2 will test the hypothesis that the pediatric airway is distinctly susceptible to pulmonary injury from hazardous VOCs, and that airway responses are modulated by extracellular vesicle (EV)-mediated signaling. The research team brings together a toxicologist, a physician- scientist and an atmospheric chemist to address the following specific aims. Aim 1 prioritizes the evaluation of 20 individual Superfund-priority VOCs to test for asthma-related phenotypes in vitro, first using a respiratory epithelial cell line (16HBE) cultured at air-liquid interface, and then in a population-based, age-appropriate model comprised of pediatric bronchial epithelial cells from the Developing Lung Molecular Atlas Program. Next, representative designed mixtures matching environmentally-relevant proportions of chemicals in ambient air will be evaluated in the standard and population-based pediatric cell lines. These responses will inform aim 2 mechanistic studies, which will test the hypothesis that VOC exposures alter EV protein expression, underlying respiratory dysfunction. It is known that inflammation and epithelial barrier function are mediated by exosomes, a class of secreted EVs ranging from 30 to 150 nm. In aim 2, EVs derived from 16HBE cells exposed to select VOCs/mixtures will be purified and sequenced using a high-throughput proteomics approach. Protein signatures revealed in this model will then be validated across diverse pediatric donor cell lines. Last, the functional role of VOC-exposed, cell-derived EVs will be evaluated using adoptive transfer experiments. In parallel to aims 1 and 2, aim 3 objectives will characterize VOC mixtures through mobile air monitoring across different locations in the greater Houston Area during baseline and in response to environmental disasters. Additionally, to fill in gaps in disaster-related toxicity testing, 16HBE cells will be directly exposed to ambient air onboard the mobile platform in the field, using time-resolved measurements to drive conditional sampling of different air masses. Outcomes from the studies proposed in Project 2 are highly relevant to the Superfund Program. Overall, the novel tools and findings will improve basic understanding of mechanisms underlying VOC-induced pediatric pulmonary injury and enable improved risk assessment to rapidly characterize respiratory hazards that threaten children’s health.