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High-throughput Screening for Small-molecule Modulators of Inward Rectifier Potassium Channels

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T-Screen as a tool to identify thyroid hormone receptor active compounds.

Arno C Gutleb¹, Ilonka A T M Meerts, Joost H Bergsma

¹Toxicology Section, Wageningen University, Tuinlaan 5, NL-6703 HE Wageningen, The Netherlands.

Environmental Toxicology and Pharmacology

|July 26, 2011

Summary

The T-Screen assay measures thyroid hormone activity by assessing cell proliferation. This method effectively evaluates compounds and environmental mixtures for their potential to interfere with thyroid hormone pathways.

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Area of Science:

Endocrinology
Cell Biology
Environmental Toxicology

Background:

Thyroid hormones (T3 and T4) are crucial for cellular functions.
Existing assays have limitations in studying thyroid hormone interference.
A need exists for a functional assay to assess thyroid hormone receptor activity.

Purpose of the Study:

To introduce and validate the T-Screen, an in vitro bioassay for thyroid hormone activity.
To assess the agonistic and antagonistic potential of compounds and environmental samples.
To bridge the gap between isolated molecule assays and complex in vivo studies.

Main Methods:

Utilized the GH3 rat pituitary tumor cell line in serum-free medium.
Measured thyroid hormone-dependent cell proliferation.
Tested compounds and extracts in the presence and absence of thyroid hormone (T3).

Main Results:

Thyroid hormones (T3, T4) and analogs (Triac, Tetrac) demonstrated dose-dependent cell growth.
Rank order of potency: Triac > T(3) > Tetrac > T(4), correlating with nuclear receptor affinities.
5,5'-diphenylhydantoin and Fentinchloride inhibited T3-induced cell growth.
Environmental extracts showed varied effects on cell proliferation, with some inhibiting and others stimulating growth.

Conclusions:

The T-Screen is a rapid and functional assay for evaluating thyroid hormone receptor activity.
It can assess the potency of pure chemicals and complex environmental mixtures.
The assay provides insights into potential endocrine disruption by environmental contaminants.