Data from: High-throughput H295R steroidogenesis assay: utility as an alternative and a statistical approach to characterize effects on steroidogenesis

The U.S. Environmental Protection Agency Endocrine Disruptor Screening Program and the Organization for Economic Co-operation and Development (OECD) have used the human adrenocarcinoma (H295R) cell-based assay to predict chemical perturbation of androgen and estrogen production. Recently, a high-throughput H295R (HT-H295R) assay was developed as part of the ToxCast program that includes measurement of 11 hormones, including progestagens, glucocorticoids, androgens, and estrogens. To date, 2012 chemicals have been screened at one concentration; of these, 656 chemicals have been screened in concentration-response. The objectives of this work were to: 1) develop an integrated analysis of chemical-mediated effects on steroidogenesis in the HT-H295R assay; and, 2) evaluate whether the HT-H295R assay predicts estrogen and androgen production specifically via comparison with the OECD-validated H295R assay. To support application of HT-H295R assay data to weight-of-evidence and prioritization tasks, a single numeric value based on Mahalanobis distances was computed for 654 chemicals to indicate the magnitude of effects on the synthesis of 11 hormones. The maximum mean Mahalanobis distance (maxmMd) values were high for strong modulators (prochloraz, mifepristone) and lower for moderate modulators (atrazine, molinate). Twenty-five of 28 reference chemicals used for OECD validation were screened in the HT-H295R assay, and produced qualitatively similar results, with accuracies of 0.90/0.75 and 0.81/0.91 for increased/decreased testosterone and estradiol production, respectively. The HT-H295R assay provides robust information regarding estrogen and androgen production, as well as additional hormones. The maxmMd from this integrated analysis may provide a data-driven approach to prioritizing lists of chemicals for putative effects on steroidogenesis.

美国环境保护署内分泌干扰物筛查计划与经济合作与发展组织（OECD）曾依托基于人肾上腺皮质癌细胞（H295R）的细胞实验，对化学物质干扰雄激素与雌激素生成的效应进行预测。近期，作为ToxCast计划的组成部分，研究人员开发了高通量H295R（HT-H295R）实验体系，该体系可同时检测包括孕激素、糖皮质激素、雄激素及雌激素在内的11种激素水平。截至目前，已有2012种化学物质完成单浓度筛查；其中656种化学物质完成了浓度-反应关系筛查。本研究的目标有二：其一，对HT-H295R实验体系中化学物质介导的类固醇生成影响开展整合分析；其二，通过与经经济合作与发展组织（OECD）验证的传统H295R实验进行对比，评估HT-H295R实验是否可特异性预测雄激素与雌激素的生成情况。为支撑HT-H295R实验数据在证据权重法与化学物质优先级排序任务中的应用，研究人员针对654种化学物质计算了基于马氏距离（Mahalanobis distances）的单一量化指标，用以表征其对11种激素合成的影响强度。平均最大马氏距离（maxmMd）值在强效调节剂（如咪鲜胺、米非司酮）中表现较高，而在中度调节剂（如阿特拉津、禾草丹）中则相对较低。用于经合组织验证的28种参考化学物质中，有25种在HT-H295R实验体系中完成了检测，所得结果定性一致；其对睾酮与雌二醇生成的促进/抑制预测准确率分别为0.90/0.75与0.81/0.91。HT-H295R实验体系可提供关于雄激素、雌激素及其他多种激素生成的可靠实验数据。本整合分析得到的平均最大马氏距离（maxmMd）指标，可为基于数据驱动的类固醇生成潜在影响化学物质优先级排序提供科学可行的方法。