کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5861270 | 1562713 | 2016 | 10 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Identification of compounds that modulate retinol signaling using a cell-based qHTS assay Identification of compounds that modulate retinol signaling using a cell-based qHTS assay](/preview/png/5861270.png)
- We developed a reporter gene assay to identify modulators of retinol signaling.
- The assay was validated in a qHTS format using LOPAC.
- Potential agonists and antagonists were identified.
- Active compounds were verified in mouse pluripotent cells.
- This assay can be used to screen large compound libraries.
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.
Journal: Toxicology in Vitro - Volume 32, April 2016, Pages 287-296