Identification of thyroid hormone receptor active compounds using a quantitative high-throughput screening platform

Jaime Freitas, Nicole Miller, Brenda J. Mengeling, Menghang Xia, Ruili Huang, Keith Houck, Ivonne M.C.M. Rietjens, John Furlow, Albertinka J. Murk

Research output: Contribution to journalArticle

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Abstract

To adapt the use of GH3.TRE-Luc reporter gene cell line for a quantitative high-throughput screening (qHTS) platform, we miniaturized the reporter gene assay to a 1536-well plate format. 1280 chemicals from the Library of Pharmacologically Active Compounds (LOPAC) and the National Toxicology Program (NTP) 1408 compound collection were analyzed to identify potential thyroid hormone receptor (TR) agonists and antagonists. Of the 2688 compounds tested, eight scored as potential TR agonists when the positive hit cut-off was defined at ≥10% efficacy, relative to maximal triiodothyronine (T3) induction, and with only one of those compounds reaching ≥20% efficacy. One common class of compounds positive in the agonist assays were retinoids such as all-trans retinoic acid, which are likely acting via the retinoid-X receptor, the heterodimer partner with the TR. Five potential TR antagonists were identified, including the antiallergy drug tranilast and the anxiolytic drug SB 205384 but also some cytotoxic compounds like 5-fluorouracil. None of the inactive compounds were structurally related to T3, nor had been reported elsewhere to be thyroid hormone disruptors, so false negatives were not detected. None of the low potency (>100 M) TR agonists resembled T3 or T4, thus these may not bind directly in the ligand-binding pocket of the receptor. For TR agonists, in the qHTS, a hit cut-off of ≥20% efficacy at 100 μM may avoid identification of positives with low or no physiological relevance. The miniaturized GH3.TRE-Luc assay offers a promising addition to the in vitro test battery for endocrine disruption, and given the low percentage of compounds testing positive, its high-throughput nature is an important advantage for future toxicological screening.

Original languageEnglish (US)
Pages (from-to)36-46
Number of pages11
JournalCurrent Chemical Genomics and Translational Medicine
Volume8
Issue number1
DOIs
StatePublished - Jan 1 2014

Fingerprint

Thyroid Hormone Receptors
Screening
Throughput
Hormones
Assays
Reporter Genes
Toxicology
Genes
Small Molecule Libraries
Retinoid X Receptors
Hormone Antagonists
Anti-Anxiety Agents
Retinoids
Triiodothyronine
Tretinoin
Thyroid Hormones
Fluorouracil
Pharmaceutical Preparations
Cells
Ligands

Keywords

  • Endocrine disruption
  • Pituitary cells
  • Quantitative high-throughput screening
  • Reporter gene assay
  • Retinoid-X receptor
  • Thyroid hormone receptor

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Identification of thyroid hormone receptor active compounds using a quantitative high-throughput screening platform. / Freitas, Jaime; Miller, Nicole; Mengeling, Brenda J.; Xia, Menghang; Huang, Ruili; Houck, Keith; Rietjens, Ivonne M.C.M.; Furlow, John; Murk, Albertinka J.

In: Current Chemical Genomics and Translational Medicine, Vol. 8, No. 1, 01.01.2014, p. 36-46.

Research output: Contribution to journalArticle

Freitas, Jaime ; Miller, Nicole ; Mengeling, Brenda J. ; Xia, Menghang ; Huang, Ruili ; Houck, Keith ; Rietjens, Ivonne M.C.M. ; Furlow, John ; Murk, Albertinka J. / Identification of thyroid hormone receptor active compounds using a quantitative high-throughput screening platform. In: Current Chemical Genomics and Translational Medicine. 2014 ; Vol. 8, No. 1. pp. 36-46.
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