Monitoring Endoplasmic Reticulum Stress Caused by Chronic Exposure to Chemicals

摘要：项目5 危险废物场地包含各种有毒化学物质的复杂混合物，这些化学物质会污染和 徘徊在环境中。许多超级基金(SF)化学品的急性毒性已被广泛 已进行了调查；然而，需要进一步研究以确定其对人类健康的慢性影响。几个SF 在尤罗克部落的环境样本中发现的化学品(例如，萘、多氯联苯和四氯化碳) 克拉马斯河流域和加利福尼亚州的空气中，已经被证明可以诱导内质网(ER)应激 在培养的细胞中。此外，在动物模型中，长期暴露于CCl4会导致组织中的内质网应激， 导致纤维化和器官损伤。因此，该项目的中心假设是，长期接触 外源物质导致内质网应激，进而导致慢性炎症、组织纤维化和最终的器官 失败了。 基于内质网应激的大小与慢性应激量成正比的新概念 接触化学物质和监测内质网应激将有助于预测由此产生的生物影响，这是长期目标 该项目的目的是开发一种高含量和中通量的生物检测方法，以测试SF化学品的潜力 诱导内质网应激(AIM IIIA)，以及用于生物液分析的内质网应激相关生物效应的生物标记物 (AIM IIIB)。为了实现这些目标，基于细胞的分析(目标I)将被用来通过 暴露在环境毒素下会导致内质网应激。此外，在动物模型(AIM II，项目4)中， 项目5将评估长期接触危险化学品对内质网压力的影响，并测试是否严重 氧脂是内质网应激的替代生物标志物(核心A和B)。开发的方法和数据 从细胞培养和动物模型中获得的信息将直接转化为开发生物标志物分析 (目标三A和B；项目2和3，以及核心A和B)。最后，研究成果将被用于服务 通过测试从克拉马斯河流域、中央山谷和 加利福尼亚州的内华达山脉山麓，以及美国各地的SF-Site或其周围(Aim IIIC with Project 1， 核心B、C和D)，以及移交给尤罗克部落环境方案的科学家(与 核心E)。 因此，该项目的总体目标是：1)测试作为潜在诊断生物标志物的脂质介体 对于经常导致器官损伤的内质网应激反应的规模，2)发展迅速，成本低廉 以及可靠的基于细胞的生物检测方法，用于检测、评估和量化有害物质对 内质网应激，3)为长期接触SF化学品对内质网应激的影响提供新的机械性见解 和纤维化疾病，4)开发生物体液的生物标记物分析，以量化组织特异性影响 外源生物对内质网应激的影响，以及5)通过分析领域，通过评估对人类健康的风险来翻译我们的发现 样本。

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.