| Development and Implementation of a High Content Nuclear Receptor Nuclear Translocation Assay Panel to Drug Discovery |
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Presentation: 4B03 Session: Imaging and High Content Screening: Valuable Tools Equally Embraced by Academia & Drug Screeners - Part 1Taosheng Chen, Yingjie Zhu, Janet Kolb, Peg Prack, Rolf Ryseck, and Michele Agler, Bristol-Myers Squibb Presenting Author: Taosheng Chen, Bristol-Myers Squibb Comapny - USA Ligand-induced cytoplasm-to-nucleus translocation is critical in nuclear receptor (NR) signal transduction. The development of fluorescent protein fusions with NRs, along with the recent developments in automated cellular imaging technology, has provided a powerful tool to monitor and quantify NR nuclear translocation. After overcoming multiple challenges unique to NRs, we have developed a robust high throughput assay system for a panel of NRs. Using a high content imaging system ligand-induced and dose-dependent NR nuclear translocation has been quantified. We have used this NR panel to study compound mechanism of action. Correlation between the translocation assay and other conventional assays (e.g., receptor binding) has been established. In addition, receptor selectivity and compound activity unavailable from conventional HTS assays was identified. A strategy to study translocation of constitutively nucleus-localized NRs, as well as the overall value of high content assays in lead identification, lead optimization and prediction of drug-drug interactions, will be discussed. |