Project 2

项目 2 摘要 项目 2 是一个新的生物医学研究项目，旨在开发新颖的工具来快速表征儿科 接触有害挥发性有机化合物 (VOC) 会带来呼吸道健康风险。这项工作将是一个 德克萨斯农工大学超级基金研究中心总体战略中的关键要素 并管理与暴露于环境紧急情况动员的危险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代表的发育阶段 尽管这些因素（即年龄、性别、种族、 和遗传学）对于哮喘风险至关重要。此外，单个/组合 VOC 的作用机制 对哮喘的发病机制知之甚少。支持有害挥发性有机化合物的评估，包括真实的挥发性有机化合物 项目 2 将检验儿童气道是城市混合体并阐明机械联系的假设 明显容易受到有害挥发性有机化合物的肺部损伤，并且气道反应受到以下因素的调节 细胞外囊泡（EV）介导的信号传导。研究小组汇集了一名毒理学家、一名医生—— 科学家和大气化学家来解决以下具体目标。目标 1 优先评估 20 个单独的超级基金优先 VOC，用于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)，然后在基于人群的、适合年龄的模型中 由来自发育肺分子图谱计划的小儿支气管上皮细胞组成。接下来， 与环境空气中化学品的环境相关比例相匹配的代表性设计混合物将 在标准和基于群体的儿科细胞系中进行评估。这些回应将为目标 2 提供信息 机制研究，将检验 VOC 暴露改变 EV 蛋白表达的假设， 呼吸功能障碍。众所周知，炎症和上皮屏障功能是由外泌体介导的， 一类分泌型 EV，波长范围为 30 至 150 nm。在目标 2 中，来自暴露于选择的 16HBE 细胞的 EV 挥发性有机化合物/混合物将使用高通量蛋白质组学方法进行纯化和测序。蛋白质特征 该模型中揭示的内容随后将在不同的儿科供体细胞系中得到验证。最后，功能作用 将使用过继转移实验来评估暴露于 VOC 的细胞衍生电动汽车。与目标 1 和 2、目标 3 将通过移动空气监测来表征 VOC 混合物的不同地点 大休斯顿地区在基线期间和应对环境灾害。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 在现场，使用时间分辨测量来驱动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。总的来说，新颖的工具和 研究结果将提高对 VOC 引起的儿童肺损伤机制的基本了解 并改进风险评估，以快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.