Evaluation of a panel of in vitro methods for assessing thyroid receptor β and transthyretin transporter disrupting activities

Reprod Toxicol. 2020 Sep:96:432-444. doi: 10.1016/j.reprotox.2019.05.011. Epub 2019 May 8.

Abstract

We developed a thyroid testing panel to assess endocrine disrupting chemicals (EDCs) capacities to bind either the thyroid receptor β (TRβ) or the thyroid hormones transporter transthyretin (TTR). We first stably transfected a human U2OS cell line with TRβ and a luciferase reporter construct to develop the TRβ CALUX® reporter gene assay to assess chemicals' potential to interact with TRβ. Secondly, we combined a TTR-binding assay with the TRβ CALUX (TTR-TRβ CALUX) and optimized the system to evaluate the competitive properties of EDCs towards T4 for TTR binding. Both systems were evaluated with a range of known thyroid-disrupting compounds. The agonistic/antagonistic TRβ CALUX successfully predicted 9/9 and 9/12 test compounds, respectively. The TTR-TRβ CALUX predicted 9/9 compounds and demonstrated competitive activities when analyzing waste water samples. We concluded that the proposed test battery is a promising screening method able to efficiently generate data on thyroid hormone interferences by chemicals.

Keywords: Bioassay; CALUX; Endocrine disruption; TTR binding potency; Thyroid hormones; Water extract.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding, Competitive
  • Biological Assay*
  • Cell Line
  • Endocrine Disruptors / pharmacology*
  • Genes, Reporter
  • Humans
  • Luciferases / genetics
  • Prealbumin / genetics
  • Prealbumin / metabolism*
  • Thyroid Hormone Receptors beta / agonists
  • Thyroid Hormone Receptors beta / antagonists & inhibitors
  • Thyroid Hormone Receptors beta / genetics
  • Thyroid Hormone Receptors beta / metabolism*
  • Thyroid Hormones / metabolism*

Substances

  • Endocrine Disruptors
  • Prealbumin
  • TTR protein, human
  • Thyroid Hormone Receptors beta
  • Thyroid Hormones
  • Luciferases