Harmonized Cross-Species Assessment of Endocrine and Metabolic Disruptors by Ecotox FACTORIAL Assay.
Harmonized Cross-Species Assessment of Endocrine and Metabolic Disruptors by Ecotox FACTORIAL Assay.
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通过 Ecotox 因子分析对内分泌和代谢干扰物进行统一的跨物种评估。
DOI：
10.1021/acs.est.0c03375
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发表时间：
2020
影响因子：
11.4
通讯作者：
Makarov,SergeiS
中科院分区：
文献类型：
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作者：
Medvedev,AlexanderV;Medvedeva,LyubovA;Martsen,Elena;Moeser,Mathew;Gorman,KristenL;Lin,Benjamin;Blackwell,Brett;Villeneuve,DanielL;Houck,KeithA;Crofton,KevinM;Makarov,SergeiS
环境污染是对人类和野生动物物种的威胁。特别值得关注的是内分泌干扰化学品（EDCs）。EDCs的一个重要靶点是通过转录调控控制内分泌和代谢反应的核受体（NRs）。由于nrr的结构差异，EDCs的不良影响在不同物种之间存在显著差异。在这里，我们描述了一种多重报告试验（Ecotox FACTORIAL），可以平行评估化合物对代表性哺乳动物、鸟类、爬行动物、两栖动物和鱼类的雌激素、雄激素、甲状腺和PPARγ受体的影响。Ecotox FACTORIAL是一种单孔试验，包括一组物种特异性的单杂交GAL4-NR报告基因构建物，瞬时转染到测试细胞中。为了协调跨物种评估，我们结合使用了两种方法。首先，我们对所有报告者使用相同类型的测试细胞；其次，我们实现了对报告rna的平行检测。该分析具有优良的质量、重现性和不显著的分析内变异性。重要的是，NR配体的EC50值与传统检测报告的值一致。使用该分析可以对不同物种的环境污染物（如双酚类、多环芳烃和合成孕激素）的潜在危害进行排序。此外，该试验允许检测地表水样品的类群特异性效应。因此，Ecotox FACTORIAL能够对化学品和地表水对人类以及野生动物的内分泌和代谢干扰活动进行统一评估。
Environmental pollution is a threat to humans and wildlife species. Of particular concern are endocrine disrupting chemicals (EDCs). An important target of EDCs is nuclear receptors (NRs) that control endocrine and metabolic responses through transcriptional regulation. Owing in part to structural differences of NRs, adverse effects of EDCs vary significantly among species. Here, we describe a multiplexed reporter assay (the Ecotox FACTORIAL) enabling parallel assessment of compounds’ effects on estrogen, androgen, thyroid, and PPARγ receptors of representative mammals, birds, reptiles, amphibians, and fish. The Ecotox FACTORIAL is a single-well assay comprising a set of species-specific, one-hybrid GAL4-NR reporter constructs transiently transfected into test cells. To harmonize cross-species assessments, we used a combination of two approaches. First, we used the same type of test cells for all reporters; second, we implemented a parallel detection of reporter RNAs. The assay demonstrated excellent quality, reproducibility, and insignificant intra-assay variability. Importantly, the EC50 values for NR ligands were consistent with those reported for conventional assays. Using the assay allowed ranking the hazard potential of environmental pollutants (e.g., bisphenols, polycyclic aromatic hydrocarbons, and synthetic progestins) across species. Furthermore, the assay permitted detecting taxa-specific effects of surface water samples. Therefore, the Ecotox FACTORIAL enables harmonized assessment of the endocrine and metabolic disrupting activity of chemicals and surface water in humans as well as in wildlife species.