Research Resource: Modulators of Glucocorticoid Receptor Activity Identified by a New High-Throughput Screening Assay

被引:7
|
作者
Blackford, John A., Jr. [1 ]
Brimacombe, Kyle R. [4 ]
Dougherty, Edward J. [1 ]
Pradhan, Madhumita [1 ]
Shen, Min [4 ]
Li, Zhuyin [4 ]
Auld, Douglas S. [4 ]
Chow, Carson C. [2 ,3 ]
Austin, Christopher P. [4 ]
Simons, S. Stoney, Jr. [1 ]
机构
[1] NIDDK, Steroid Hormones Sect, NIH, Bethesda, MD 20892 USA
[2] NIDDK, Lab Endocrinol & Receptor Biol, NIH, Bethesda, MD 20892 USA
[3] NIDDK, Lab Biol Modeling, NIH, Bethesda, MD 20892 USA
[4] NIH, Natl Ctr Adv Translat Sci, Rockville, MD 20892 USA
基金
美国国家卫生研究院;
关键词
SMALL-MOLECULE INHIBITOR; MEDIATED GENE INDUCTION; DOSE-RESPONSE CURVE; ESTROGEN-RECEPTOR; COACTIVATOR BINDING; COREPRESSOR BINDING; ANDROGEN RECEPTOR; DISORDERED AF1; TRANSACTIVATION; AGONISTS;
D O I
10.1210/me.2014-1069
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
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 EC50 (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 EC50 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.
引用
收藏
页码:1194 / 1206
页数:13
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