Novel assays monitoring direct glucocorticoid receptor protein activity exhibit high predictive power for ligand activity on endogenous gene targets
- Author
- Laura Van Moortel (UGent) , Jonathan Thommis (UGent) , Brecht Maertens (UGent) , An Staes (UGent) , Dorien Clarisse (UGent) , Delphine De Sutter (UGent) , Claude Libert (UGent) , Onno C. Meijer, Sven Eyckerman (UGent) , Kris Gevaert (UGent) and Karolien De Bosscher (UGent)
- Organization
- Project
- Abstract
- Exogenous glucocorticoids are widely used in the clinic for the treatment of inflammatory disorders and auto immune diseases. Unfortunately, their use is hampered by many side effects and therapy resistance. Efforts to find more selective glucocorticoid receptor (GR) agonists and modulators (called SEGRAMs) that are able to separate anti-inflammatory effects via gene repression from metabolic effects via gene activation, have been unsuccessful so far. In this study, we characterized a set of functionally diverse GR ligands in A549 cells, first using a panel of luciferase-based reporter gene assays evaluating GR-driven gene activation and gene repression. We expanded this minimal assay set with novel luciferase-based read-outs monitoring GR protein levels, GR dimerization and GR Serine 211 (Ser211) phosphorylation status and compared their outcomes with compound effects on the mRNA levels of known GR target genes in A549 cells and primary hepatocytes. We found that luciferase reporters evaluating GR-driven gene activation and gene repression were not always reliable predictors for effects on endogenous target genes. Remarkably, our novel assay monitoring GR Ser211 phosphorylation levels proved to be the most reliable predictor for compound effects on almost all tested endogenous GR targets, both driven by gene activation and repression. The integration of this novel assay in existing screening platforms running both in academia and industry may therefore boost chances to find novel GR ligands with an actual improved therapeutic benefit.
- Keywords
- Pharmacology, General Medicine, Glucocorticoids, Glucocorticoid receptor, SEGRAM, Drug discovery, Inflammation, Assay development, FACTOR-KAPPA-B, ACTIVATOR PROTEIN-1, BINDING, TRANSCRIPTION, PHOSPHORYLATION, MECHANISMS, DIMERIZATION, SEQUENCE, AGONIST, HEALTH
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-8761100
- MLA
- Van Moortel, Laura, et al. “Novel Assays Monitoring Direct Glucocorticoid Receptor Protein Activity Exhibit High Predictive Power for Ligand Activity on Endogenous Gene Targets.” BIOMEDICINE & PHARMACOTHERAPY, vol. 152, 2022, doi:10.1016/j.biopha.2022.113218.
- APA
- Van Moortel, L., Thommis, J., Maertens, B., Staes, A., Clarisse, D., De Sutter, D., … De Bosscher, K. (2022). Novel assays monitoring direct glucocorticoid receptor protein activity exhibit high predictive power for ligand activity on endogenous gene targets. BIOMEDICINE & PHARMACOTHERAPY, 152. https://doi.org/10.1016/j.biopha.2022.113218
- Chicago author-date
- Van Moortel, Laura, Jonathan Thommis, Brecht Maertens, An Staes, Dorien Clarisse, Delphine De Sutter, Claude Libert, et al. 2022. “Novel Assays Monitoring Direct Glucocorticoid Receptor Protein Activity Exhibit High Predictive Power for Ligand Activity on Endogenous Gene Targets.” BIOMEDICINE & PHARMACOTHERAPY 152. https://doi.org/10.1016/j.biopha.2022.113218.
- Chicago author-date (all authors)
- Van Moortel, Laura, Jonathan Thommis, Brecht Maertens, An Staes, Dorien Clarisse, Delphine De Sutter, Claude Libert, Onno C. Meijer, Sven Eyckerman, Kris Gevaert, and Karolien De Bosscher. 2022. “Novel Assays Monitoring Direct Glucocorticoid Receptor Protein Activity Exhibit High Predictive Power for Ligand Activity on Endogenous Gene Targets.” BIOMEDICINE & PHARMACOTHERAPY 152. doi:10.1016/j.biopha.2022.113218.
- Vancouver
- 1.Van Moortel L, Thommis J, Maertens B, Staes A, Clarisse D, De Sutter D, et al. Novel assays monitoring direct glucocorticoid receptor protein activity exhibit high predictive power for ligand activity on endogenous gene targets. BIOMEDICINE & PHARMACOTHERAPY. 2022;152.
- IEEE
- [1]L. Van Moortel et al., “Novel assays monitoring direct glucocorticoid receptor protein activity exhibit high predictive power for ligand activity on endogenous gene targets,” BIOMEDICINE & PHARMACOTHERAPY, vol. 152, 2022.
@article{8761100, abstract = {{Exogenous glucocorticoids are widely used in the clinic for the treatment of inflammatory disorders and auto immune diseases. Unfortunately, their use is hampered by many side effects and therapy resistance. Efforts to find more selective glucocorticoid receptor (GR) agonists and modulators (called SEGRAMs) that are able to separate anti-inflammatory effects via gene repression from metabolic effects via gene activation, have been unsuccessful so far. In this study, we characterized a set of functionally diverse GR ligands in A549 cells, first using a panel of luciferase-based reporter gene assays evaluating GR-driven gene activation and gene repression. We expanded this minimal assay set with novel luciferase-based read-outs monitoring GR protein levels, GR dimerization and GR Serine 211 (Ser211) phosphorylation status and compared their outcomes with compound effects on the mRNA levels of known GR target genes in A549 cells and primary hepatocytes. We found that luciferase reporters evaluating GR-driven gene activation and gene repression were not always reliable predictors for effects on endogenous target genes. Remarkably, our novel assay monitoring GR Ser211 phosphorylation levels proved to be the most reliable predictor for compound effects on almost all tested endogenous GR targets, both driven by gene activation and repression. The integration of this novel assay in existing screening platforms running both in academia and industry may therefore boost chances to find novel GR ligands with an actual improved therapeutic benefit.}}, articleno = {{113218}}, author = {{Van Moortel, Laura and Thommis, Jonathan and Maertens, Brecht and Staes, An and Clarisse, Dorien and De Sutter, Delphine and Libert, Claude and Meijer, Onno C. and Eyckerman, Sven and Gevaert, Kris and De Bosscher, Karolien}}, issn = {{0753-3322}}, journal = {{BIOMEDICINE & PHARMACOTHERAPY}}, keywords = {{Pharmacology,General Medicine,Glucocorticoids,Glucocorticoid receptor,SEGRAM,Drug discovery,Inflammation,Assay development,FACTOR-KAPPA-B,ACTIVATOR PROTEIN-1,BINDING,TRANSCRIPTION,PHOSPHORYLATION,MECHANISMS,DIMERIZATION,SEQUENCE,AGONIST,HEALTH}}, language = {{eng}}, pages = {{15}}, title = {{Novel assays monitoring direct glucocorticoid receptor protein activity exhibit high predictive power for ligand activity on endogenous gene targets}}, url = {{http://doi.org/10.1016/j.biopha.2022.113218}}, volume = {{152}}, year = {{2022}}, }
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