2. Academic Validation
  3. Extended steroid profiling in H295R cells provides deeper insight into chemical-induced disturbances of steroidogenesis: Exemplified by prochloraz and anabolic steroids

Extended steroid profiling in H295R cells provides deeper insight into chemical-induced disturbances of steroidogenesis: Exemplified by prochloraz and anabolic steroids

  • Mol Cell Endocrinol. 2023 Apr 8;111929. doi: 10.1016/j.mce.2023.111929.
Marie-Christin Jäger 1,  Melanie Patt 2,  Víctor González-Ruiz 3,  Julien Boccard 4,  Tim Wey 5,  Denise V Winter 6,  Serge Rudaz 7,  Alex Odermatt 8
Affiliations

Affiliations

  • 1 Swiss Centre for Applied Human Toxicology (SCAHT), University of Basel, Missionsstrasse 64, 4055, Basel, Switzerland; Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, Klingelbergstrasse 50, 4056, Basel, Switzerland. Electronic address: [email protected].
  • 2 Swiss Centre for Applied Human Toxicology (SCAHT), University of Basel, Missionsstrasse 64, 4055, Basel, Switzerland; Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, Klingelbergstrasse 50, 4056, Basel, Switzerland. Electronic address: [email protected].
  • 3 Swiss Centre for Applied Human Toxicology (SCAHT), University of Basel, Missionsstrasse 64, 4055, Basel, Switzerland; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva 4, Switzerland. Electronic address: [email protected].
  • 4 Swiss Centre for Applied Human Toxicology (SCAHT), University of Basel, Missionsstrasse 64, 4055, Basel, Switzerland; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva 4, Switzerland. Electronic address: [email protected].
  • 5 Swiss Centre for Applied Human Toxicology (SCAHT), University of Basel, Missionsstrasse 64, 4055, Basel, Switzerland; Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, Klingelbergstrasse 50, 4056, Basel, Switzerland. Electronic address: [email protected].
  • 6 Swiss Centre for Applied Human Toxicology (SCAHT), University of Basel, Missionsstrasse 64, 4055, Basel, Switzerland; Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, Klingelbergstrasse 50, 4056, Basel, Switzerland. Electronic address: [email protected].
  • 7 Swiss Centre for Applied Human Toxicology (SCAHT), University of Basel, Missionsstrasse 64, 4055, Basel, Switzerland; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva 4, Switzerland. Electronic address: [email protected].
  • 8 Swiss Centre for Applied Human Toxicology (SCAHT), University of Basel, Missionsstrasse 64, 4055, Basel, Switzerland; Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, Klingelbergstrasse 50, 4056, Basel, Switzerland. Electronic address: [email protected].
PMID: 37037411 DOI: 10.1016/j.mce.2023.111929
Abstract

Human adrenocortical H295R cells have been validated by the OECD Test Guideline 456 to detect chemicals disrupting testosterone and 17β-estradiol (estradiol) biosynthesis. This study evaluated a novel approach to detect disturbances of steroidogenesis in H295R cells, exemplified by prochloraz and five anabolic Steroids. Steroid profiles were assessed by an untargeted LC-MS-based method, providing a relative quantification of 57 Steroids annotated according to their accurate masses and retention times. Such a panel of Steroids included several mineralocorticoids, glucocorticoids, progestins and adrenal androgens. The coverage of a high number of metabolites in this extended steroid profiling facilitated grouping of chemicals with similar effects and detecting subtler differences between chemicals. It allowed, for example, distinguishing between the effects of turinabol and oxymetholone, supposed to act similarly in a previous characterization including only nine adrenal Steroids. Furthermore, the results revealed that product/substrate ratios can provide superior information on altered Enzyme activities compared to individual metabolite levels. For example, the 17α-hydroxypregnenolone/pregnenolone ratio was found to be a more sensitive marker for detecting 17α-hydroxylase inhibition by prochloraz than the corresponding individual Steroids. These results illustrate that chemical grouping and calculation of product/substrate ratios can provide valuable information on mode-of-action and help prioritizing further experimental work.

Keywords

Adrenal; Anabolic steroid; Androgen; H295R; LC-MS; Steroid profiling; Steroidogenesis.

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