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（U.S. Environmental Protection Agency）与经济合作与发展组织（Organization for Economic Co-operation and Development, OECD）曾采用人肾上腺皮质癌细胞（human adrenocarcinoma, H295R）细胞测定法，预测化学物对雄激素与雌激素生成的干扰效应。近期，作为ToxCast项目的组成部分，高通量H295R（high-throughput H295R, HT-H295R）细胞测定法被开发问世，该方法可同时检测11种激素，涵盖孕激素、糖皮质激素、雄激素与雌激素四类。截至目前，已有2012种化学物在单一浓度下完成筛选；其中656种化学物完成了浓度-反应关系筛选实验。
本研究的目标为：1）对HT-H295R细胞测定法中化学物介导的类固醇生成影响开展整合分析；2）通过与经经合组织验证的H295R细胞测定法对比，评估HT-H295R细胞测定法是否可特异性预测雄激素与雌激素的生成。
为支持将HT-H295R细胞测定法数据应用于证据权重与优先级排序任务，研究人员针对654种化学物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。强调节剂丙环唑（prochloraz）、米非司酮（mifepristone）的最大平均马氏距离（maxmMd）数值较高，而中度调节剂阿特拉津（atrazine）、禾草丹（molinate）的该数值则相对较低。
用于经合组织验证的28种参考化学物中，有25种在HT-H295R细胞测定法中完成了筛选，所得结果具备定性一致性；其对睾酮与雌二醇生成的促进/抑制作用的准确率分别为0.90/0.75与0.81/0.91。HT-H295R细胞测定法可提供关于雄激素、雌激素及其他多种激素生成的稳健信息。本整合分析得到的最大平均马氏距离值，可为针对疑似具有类固醇生成干扰效应的化学物列表开展数据驱动式优先级排序提供可行方法。