Monitoring Endoplasmic Reticulum Stress Caused by Chronic Exposure to Chemicals

摘要：项目5 危险废物场含有各种有毒化学品的复杂混合物， 在环境中徘徊。许多超级基金（SF）化学品的急性毒性已被广泛 然而，需要进一步研究，以确定其对人类健康的长期影响。几个SF 化学物质（如萘，多氯联苯和四氯化碳），这是从尤罗克部落的环境样本中发现的 在克拉马斯河流域和加州空气中，已经证明诱导内质网（ER）应激 在培养的细胞中。此外，在动物模型中，长期暴露于CCl 4导致组织中的ER应激， 导致纤维化和器官损伤。因此，该项目的中心假设是，长期暴露于 外源性物质导致ER应激，然后导致慢性炎症、组织纤维化和最终的器官损伤。 失败 基于ER应激的大小与慢性炎症的量成正比的新概念， 暴露于化学物质，并监测ER压力将有助于预测由此产生的生物效应， 本项目的主要目的是开发一种高含量和中等通量的生物测定方法，以测试SF化学品的潜力 诱导ER应激（目的IIIA），以及用于生物流体分析的ER应激相关生物效应的生物标志物 (Aim IIIB）。为了实现这些目标，将使用基于细胞的测定（Aim I）来理解机制， 暴露于环境毒素会导致内质网应激此外，在动物模型中（Aim II，项目4）， 项目5将评估长期暴露于危险化学品对ER应激的影响，并测试是否存在血清 含氧脂质是ER应激的替代生物标志物（具有核心A和B）。制定的方法和数据 从细胞培养物和动物模型中获得的生物标志物将直接转化为生物标志物测定 (Aim IIIA和B;项目2和3以及核心A和B）。最后，研究结果将用于服务 通过测试从克拉马斯河流域、中央谷和 加州的Sierra内华达州山麓丘陵，以及美国的SF站点或其周围（Aim IIIC与项目1， 核心B，C和D），以及转让给科学家的Yurok部落环境计划（与 核心E）。 因此，本项目的总体目标是：1）测试脂质介质作为潜在的诊断生物标志物 对于经常导致器官损伤的ER应激反应的强度，2）开发快速、廉价 和可靠的新的基于细胞的生物测定，以检测，评估和定量有害物质对 内质网应激，3）提供了新的机制的见解，慢性暴露于SF化学品对内质网应激的影响 和纤维化疾病，4）开发用于生物流体的生物标志物测定，用于量化 外源性物质对内质网应激的影响，5）通过评估对人类健康的风险，通过分析领域， 样品

ABSTRACT: PROJECT 5 Hazardous waste sites contain complex mixtures of a wide variety of toxic chemicals that contaminate and linger in the environment. The acute toxicities of numerous Superfund (SF) chemicals have been extensively investigated; however, further studies are needed to determine their chronic effects on human health. Several SF chemicals (e.g. naphthalene, PCBs, and CCl4), which are found in environmental samples from the Yurok Tribe of the Klamath River basin, and in California air, have been shown to induce endoplasmic reticulum (ER) stress in cultured cells. Furthermore, in animal models, long-term exposure to CCl4 leads to ER stress in tissues, resulting in fibrosis and organ damage. Thus, the central hypothesis for this project is that chronic exposure to xenobiotics leads to ER stress that then contributes to chronic inflammation, tissue fibrosis and eventual organ failure. Based on the novel concept that the magnitude of ER stress is proportional to the amount of chronic exposure to chemicals, and monitoring ER stress will help predict resulting biological effects, the long term goal of this project is to develop a high-content and medium throughput bioassay to test the potential of SF chemicals to induce ER stress (Aim IIIA), and a biomarker of ER stress-associated biological effects for bio-fluid analysis (Aim IIIB). Toward these objectives, cell-based assays (Aim I) will be used to understand the mechanisms by which exposure to environmental toxins leads to ER stress. In addition, in animal models (Aim II, with Project 4), Project 5 will evaluate the effects of chronic exposure to hazardous chemicals on ER stress, and test if seric oxylipids are surrogate biomarkers for ER stress (with Cores A and B). The methodology developed and data obtained from the cell cultures and animal models will be directly translated in developing biomarker assays (Aim IIIA and B; with Projects 2 and 3, and Cores A and B). Finally, research findings will be utilized to serve the community at large by testing field samples collected from the Klamath River basin, the Central Valley and the Sierra Nevada foothills in California, and at or around SF-sites across the U.S. (Aim IIIC with Project 1, Cores B, C and D), as well as transferring to the scientists of the Yurok Tribe Environmental Program (with Core E). Accordingly, the overall goals of this project are to: 1) test lipid mediators as potential diagnostic biomarkers for the magnitude of ER stress response that often contributes to organ damage, 2) develop fast, inexpensive and reliable new cell-based bioassays to detect, assess and quantitate the effects of hazardous substances on ER stress, 3) provide new mechanistic insights into the effects of chronic exposure to SF chemicals on ER stress and fibrotic diseases, 4) develop biomarkers assays for bio-fluids for the quantification of tissue-specific effects of xenobiotics on ER stress, and 5) translate our findings by assessing risk on human health, by analyzing field samples.