Disruption of steroidogenesis by environmental chemicals can result in altered hormone levels causing adverse reproductive and developmental effects. A high-throughput assay using H295R human adrenocortical carcinoma cells was used to evaluate the effect of 2060 chemical samples on steroidogenesis via high-performance liquid chromatography followed by tandem mass spectrometry quantification of 10 steroid hormones, including progestagens, glucocorticoids, androgens, and estrogens. The study employed a 3 stage screening strategy. The first stage established the maximum tolerated concentration (MTC; ≥ 70% viability) per sample. The second stage quantified changes in hormone levels at the MTC whereas the third stage performed concentration-response (CR) on a subset of samples. At all stages, cells were prestimulated with 10 µM forskolin for 48 h to induce steroidogenesis followed by chemical treatment for 48 h. Of the 2060 chemical samples evaluated, 524 samples were selected for 6-point CR screening, based in part on significantly altering at least 4 hormones at the MTC. CR screening identified 232 chemical samples with concentration-dependent effects on 17β-estradiol and/or testosterone, with 411 chemical samples showing an effect on at least one hormone across the steroidogenesis pathway. Clustering of the concentration-dependent chemical-mediated steroid hormone effects grouped chemical samples into 5 distinct profiles generally representing putative mechanisms of action, including CYP17A1 and HSD3B inhibition. A distinct pattern was observed between imidazole and triazole fungicides suggesting potentially distinct mechanisms of action. From a chemical testing and prioritization perspective, this assay platform provides a robust model for high-throughput screening of chemicals for effects on steroidogenesis. This dataset is associated with the following publication: Karmaus , A., C. Toole, D. Filer , K. Lewis, and M. Martin. (Toxicological Sciences) High-throughput screening of chemical effects on steroidogenesis using H295R human adrenocortical carcinoma cells. TOXICOLOGICAL SCIENCES. Society of Toxicology, 150(2): 323-332, (2016).

HT-H295R data was downloaded using the ToxCast pipeline (tcpl) R package and is publicly available. Multi-concentration level 0 data from invitrodb (version 3.1) were downloaded and converted from g/ml into micromolar concentrations prior to calculation of mMds and data simulation (Supplemental Data 1). This dataset is associated with the following publication: Haggard, D., W. Setzer, R. Judson, and K. Friedman. Development of a prioritization method for chemical-mediated effects on steroidogenesis using an integrated statistical analysis of high-throughput H295R data. REGULATORY TOXICOLOGY AND PHARMACOLOGY. Elsevier Science Ltd, New York, NY, USA, 109: 104510, (2019).

HT-H295R data was downloaded using the ToxCast pipeline (tcpl) R package and is publicly available. Multi-concentration level 0 data from invitrodb (version 3.1) were downloaded and converted from g/ml into micromolar concentrations prior to calculation of mMds and data simulation (Supplemental Data 1). This dataset is associated with the following publication: Haggard, D., W. Setzer, R. Judson, and K. Friedman. Development of a prioritization method for chemical-mediated effects on steroidogenesis using an integrated statistical analysis of high-throughput H295R data. REGULATORY TOXICOLOGY AND PHARMACOLOGY. Elsevier Science Ltd, New York, NY, USA, 109: 104510, (2019).

Dataset from Foster, M.J., et al., Evaluating structure-based activity in a high-throughput assay for steroid biosynthesis, Computational Toxicology, Vol 24, No. 100245, Nov 2022, DOI https://doi.org/10.1016/j.comtox.2022.100245 The work described herein was conducted in R software (version 4.0.2) and is available at GitHub (https://github.com/USEPA/CompTox-HTH295R-SAR) and the US EPA Clowder repository as well as in Supplemental File 1 as an html file including code and resultant output. This dataset is associated with the following publication: Foster, M., G. Patlewicz, I. Shah, D. Haggard, R. Judson, and K. Friedman. Evaluating structure-based activity in a high-throughput assay for steroid biosynthesis. Computational Toxicology. Elsevier B.V., Amsterdam, NETHERLANDS, 24: 100245, (2022).

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. This dataset is associated with the following publication: Haggard, D., A. Karmaus, M. Martin, R. Judson, W. Setzer, and K. Paul-Friedman. (Toxicological Sciences) High-throughput H295R steroidogenesis assay: utility as an alternative and a statistical approach to characterize effects on steroidogenesis. TOXICOLOGICAL SCIENCES. Society of Toxicology, RESTON, VA, 162(2): 509-534, (2018).

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. This dataset is associated with the following publication: Haggard, D., A. Karmaus, M. Martin, R. Judson, W. Setzer, and K. Paul-Friedman. (Toxicological Sciences) High-throughput H295R steroidogenesis assay: utility as an alternative and a statistical approach to characterize effects on steroidogenesis. TOXICOLOGICAL SCIENCES. Society of Toxicology, RESTON, VA, 162(2): 509-534, (2018).

The greater dynamic range of testosterone production in a highly purified rat Leydig cell assay permitted the detection of chemical induced inhibition that was not detected by the high throughput screening format of the H295R steroidogenesis assay. This dataset is associated with ORD-022468 entered in STICS. This dataset is associated with the following publication: Botteri Principato, N., J. Suarez, S. Laws, and G. Klinefelter. The Use of Purified Rat Leydig Cells Complements the H295R Screen to Detect Chemical Induced Alterations in Testosterone Production. BIOLOGY OF REPRODUCTION. Society for the Study of Reproduction, 98(2): 239-249, (2018).

The greater dynamic range of testosterone production in a highly purified rat Leydig cell assay permitted the detection of chemical induced inhibition that was not detected by the high throughput screening format of the H295R steroidogenesis assay. This dataset is associated with ORD-022468 entered in STICS. This dataset is associated with the following publication: Botteri Principato, N., J. Suarez, S. Laws, and G. Klinefelter. The Use of Purified Rat Leydig Cells Complements the H295R Screen to Detect Chemical Induced Alterations in Testosterone Production. BIOLOGY OF REPRODUCTION. Society for the Study of Reproduction, 98(2): 239-249, (2018).

Dataset for 'Evaluation of a high-throughput H295R homogenous time resolved fluorescence assay for androgen and estrogen steroidogenesis screening', Garnovskaya, et al., Toxicology in Vitro, Vol 92, 105659, Oct 2023, https://doi.org/10.1016/j.tiv.2023.105659. Contains the ToxCast Pipeline datasets and curve fits that can be pulled up from InvitroDB on the CompTox Chemicals Dashboard. This dataset is associated with the following publication: Garnovskaya, M., M. Feshuk, W. Stewart, K. Friedman, R. Thomas, and C. Deisenroth. Evaluation of a High-throughput H295R Homogenous Time Resolved Fluorescence Assay for Androgen and Estrogen Steroidogenesis Screening. TOXICOLOGY IN VITRO. Elsevier Science Ltd, New York, NY, USA, 92: 105659, (2023).

The greater dynamic range of testosterone production in a highly purified rat Leydig cell assay permitted the detection of chemical induced inhibition that was not detected by the high throughput screening format of the H295R steroidogenesis assay. This dataset is associated with the following publication: Klinefelter , G., J. Laskey, and R. Amann. Statin Drugs Markedly Inhibit Testosterone Production by Rat Leydig Cells In Vitro: Implications for Men. REPRODUCTIVE TOXICOLOGY. Elsevier Science Ltd, New York, NY, USA, 45: 52-58, (2014). NOTE: This dataset has been removed from public access due to revocation. Please refer inquiries regarding this dataset to the listed contact person.