Research resource: modulators of glucocorticoid receptor activity identified by a new high-throughput screening assay
| Autor: | Zhuyin Li, Christopher P. Austin, John A. Blackford, S. Stoney Simons, Carson C. Chow, Madhumita Pradhan, Edward J. Dougherty, Kyle R. Brimacombe, Min Shen, Douglas S. Auld |
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| Rok vydání: | 2014 |
| Předmět: |
Transcriptional Activation
Transcription Genetic High-throughput screening Green Fluorescent Proteins Pharmacology Biology Dexamethasone Cell Line Transactivation Endocrinology Glucocorticoid receptor Receptors Glucocorticoid High-Throughput Screening Assays medicine Humans Research Resource Receptor Molecular Biology Glucocorticoids HEK 293 cells General Medicine HEK293 Cells Signal transduction Glucocorticoid medicine.drug Signal Transduction |
| Zdroj: | Molecular endocrinology (Baltimore, Md.). 28(7) |
| ISSN: | 1944-9917 |
| Popis: | Glucocorticoid steroids affect almost every type of tissue and thus are widely used to treat a variety of human pathological conditions. However, the severity of numerous side effects limits the frequency and duration of glucocorticoid treatments. Of the numerous approaches to control off-target responses to glucocorticoids, small molecules and pharmaceuticals offer several advantages. Here we describe a new, extended high-throughput screen in intact cells to identify small molecule modulators of dexamethasone-induced glucocorticoid receptor (GR) transcriptional activity. The novelty of this assay is that it monitors changes in both GR maximal activity (A(max)) and EC(50) (the position of the dexamethasone dose-response curve). Upon screening 1280 chemicals, 10 with the greatest changes in the absolute value of A(max) or EC(50) were selected for further examination. Qualitatively identical behaviors for 60% to 90% of the chemicals were observed in a completely different system, suggesting that other systems will be similarly affected by these chemicals. Additional analysis of the 10 chemicals in a recently described competition assay determined their kinetically defined mechanism and site of action. Some chemicals had similar mechanisms of action despite divergent effects on the level of the GR-induced product. These combined assays offer a straightforward method of identifying numerous new pharmaceuticals that can alter GR transactivation in ways that could be clinically useful. |
| Databáze: | OpenAIRE |
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