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Toxicol In Vitro. 2016 Apr;32:287-96. doi: 10.1016/j.tiv.2016.01.011. Epub 2016 Jan 25.

Identification of compounds that modulate retinol signaling using a cell-based qHTS assay.

Author information

1
Division of Molecular Biology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, Laurel, MD 20708, United States. Electronic address: yanling.chen@fda.hhs.gov.
2
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD 20892, United States.
3
Division of Molecular Biology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, Laurel, MD 20708, United States.

Abstract

In vertebrates, the retinol (vitamin A) signaling pathway (RSP) controls the biosynthesis and catabolism of all-trans retinoic acid (atRA), which regulates transcription of genes essential for embryonic development. Chemicals that interfere with the RSP to cause abnormal intracellular levels of atRA are potential developmental toxicants. To assess chemicals for the ability to interfere with retinol signaling, we have developed a cell-based RARE (Retinoic Acid Response Element) reporter gene assay to identify RSP disruptors. To validate this assay in a quantitative high-throughput screening (qHTS) platform, we screened the Library of Pharmacologically Active Compounds (LOPAC) in both agonist and antagonist modes. The screens detected known RSP agonists, demonstrating assay reliability, and also identified novel RSP agonists including kenpaullone, niclosamide, PD98059 and SU4312, and RSP antagonists including Bay 11-7085, LY294002, 3,4-Methylenedioxy-β-nitrostyrene, and topoisomerase inhibitors (camptothecin, topotecan, amsacrine hydrochloride, and idarubicin). When evaluated in the P19 pluripotent cell, these compounds were found to affect the expression of the Hoxa1 gene that is essential for embryo body patterning. These results show that the RARE assay is an effective qHTS approach for screening large compound libraries to identify chemicals that have the potential to adversely affect embryonic development through interference with retinol signaling.

KEYWORDS:

Developmental toxicant; High-throughput screening; LOPAC; Retinol signaling; Stem cell; Tox21

PMID:
26820057
PMCID:
PMC4779714
DOI:
10.1016/j.tiv.2016.01.011
[Indexed for MEDLINE]
Free PMC Article

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