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Towards an integrated in vitro strategy for estrogenicity testing.

Si Wang1, Jac M M J G Aarts, Laura H J de Haan

  • 1Division of Toxicology, Wageningen University and Research Centre, Tuinlaan 5, 6703 HE, Wageningen, The Netherlands; Business Unit of Toxicology & Bioassays, RIKILT - Institute of Food Safety, Wageningen University and Research Centre, Akkermaalsbos 2, 6708 WB, Wageningen, The Netherlands.

Journal of Applied Toxicology : JAT
|October 12, 2013
PubMed
Summary

This study evaluated in vitro assays for estrogenicity testing, finding that combining yeast estrogen bioassays with U2OS ERα-CALUX assays shows promise for an integrated testing strategy (ITS) to predict in vivo effects.

Keywords:
correlationestrogenicityintegrated testing strategyreporter gene assaysuterotrophic assay

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Area of Science:

  • Toxicology
  • Endocrinology
  • In Vitro Assays

Background:

  • Estrogenicity testing is crucial for chemical safety assessment.
  • Existing in vitro assays vary in their ability to predict in vivo estrogenic effects.
  • An integrated testing strategy (ITS) is needed to reliably assess estrogenicity and reduce animal testing.

Purpose of the Study:

  • To define an in vitro integrated testing strategy (ITS) for estrogenicity.
  • To compare the predictive accuracy of various in vitro assays against the in vivo uterotrophic assay.
  • To identify discrepancies between in vitro and in vivo estrogenicity testing.

Main Methods:

  • Tested 23 reference compounds across multiple in vitro assays (proliferation, reporter gene assays).
  • Integrated results with estrogen receptor (ER) binding and transcriptional activation (TA) assay data.
  • Correlated in vitro outcomes with the in vivo uterotrophic assay.

Main Results:

  • All tested in vitro assays showed good correlation (R² = 0.62–0.87) with the in vivo uterotrophic assay.
  • The combination of yeast estrogen bioassay and U2OS ERα-CALUX assay appeared most promising for an ITS.
  • Outliers included 4-hydroxytamoxifen, testosterone, apigenin, tamoxifen, and kepone.

Conclusions:

  • The combination of specific in vitro assays offers a promising approach for an estrogenicity ITS.
  • Further refinement requires incorporating assays for digestion, bioavailability, and metabolism.
  • Improving in vitro strategies can potentially replace animal testing for (anti-)estrogenic effects.