A high throughput screening assay for human Thyroperoxidase inhibitors.

Autor: Dong H; Hazard Identification Division, Environmental Health Science & Research Bureau, Health Canada, Ottawa, Ontario, Canada. Electronic address: Hongyan.Dong@hc-sc.gc.ca., Friedman KP; Center for Computational Toxicology and Exposure, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina, USA. Electronic address: Friedman.Katie@epa.gov., Filiatreault A; Hazard Identification Division, Environmental Health Science & Research Bureau, Health Canada, Ottawa, Ontario, Canada. Electronic address: Alain.Filiatreau@hc-sc.gc.ca., Thomson EM; Hazard Identification Division, Environmental Health Science & Research Bureau, Health Canada, Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, Canada. Electronic address: Errol.Thomson@hc-sc.gc.ca., Wade MG; Hazard Identification Division, Environmental Health Science & Research Bureau, Health, Department of Cellular & Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada. Electronic address: Mike.Wade@hc-sc.gc.ca.
Jazyk: angličtina
Zdroj: Toxicology in vitro : an international journal published in association with BIBRA [Toxicol In Vitro] 2024 Dec; Vol. 101, pp. 105946. Date of Electronic Publication: 2024 Sep 28.
DOI: 10.1016/j.tiv.2024.105946
Abstrakt: Rapid, human relevant assays are needed to assess potential hazards of the many chemicals in commerce. An assay of thyroid peroxidase (TPO) inhibition, using the substrate Amplex Ultra Red, was recently adapted for human TPO (AUR-hTPO). We tested a large number (788) of chemicals through this AUR-hTPO assay and compared performance with published results from an assay using enzyme from rat thyroid microsomes (AUR-rTPO). Coded chemicals, from the US EPA ToxCast Inventory, were tested in a tiered approach: 1) Initial screening at a single concentration; 2) Potency estimation for active chemicals with multiple concentrations; 3) Screening active chemicals for the non-specific activity. The assay gave consistent results for positive chemical methimazole and several positive and negative reference chemicals. hTPO inhibition was observed for 190 chemicals reported as positive in rTPO. Of these, 158 showed no confounding activity (interference due to fluorescence or non-specific protein inhibition). Comparison of all result with rTPO data and with evidence of TPO inhibition found in the literature suggest that the current assay has a higher rate of false negative but a much lower rate of false positive compared with the rTPO screen. These findings underscore the effectiveness of the AUR assay, using hTPO enzyme from engineered cell lines, to identify moderate to strong inhibitors but some improvements may be needed to detect weak TPO inhibitors.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024. Published by Elsevier Ltd.)
Databáze: MEDLINE
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