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BioAssay: AID 2551

qHTS for inhibitors of ROR gamma transcriptional activity

The retinoic acid-related orphan receptor (ROR) gamma is a transcription factor that has a central role in the differentiation of Th17 cells, a subset of T helper cells that secrete the inflammatory cytokines IL-17, IL-17F, and IL-22. Th17 cells have been implicated in graft versus host disease, autoimmune disease and asthma. ROR gamma is induced in naive T helper cells in the presence of more ..
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 Tested Compounds
 Tested Compounds
All(305448)
 
 
Active(16716)
 
 
Inactive(253872)
 
 
Inconclusive(35306)
 
 
 Tested Substances
 Tested Substances
All(309031)
 
 
Active(16824)
 
 
Inactive(256777)
 
 
Inconclusive(35430)
 
 
AID: 2551
Data Source: NCGC (RORG318)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2010-03-15

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 16716
Related Experiments
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AIDNameTypeComment
2604Quantitative high throughput screen for inhibitors of ROR gamma transcriptional activity: SummarySummarydepositor-specified cross reference
2762Concentration response confirmation assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference
489036VP16 selectivity assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference
489037Secondary confirmation assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference
489038DHR3 selectivity assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference
489039ROR alpha selectivity assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference
492954Mouse Th17 T cell differentiation assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference
492962Mouse Th1 T cell differentiation assay for inhibitors of ROR gamma transcriptional activityOtherdepositor-specified cross reference
2546VP16 counterscreen qHTS for inhibitors of ROR gamma transcriptional activityConfirmatorysame project related to Summary assay
2763Concentration response confirmation VP16 counterscreen for inhibitors of ROR gamma transcriptional activityConfirmatorysame project related to Summary assay
Description:
NIH Molecular Libraries Probe Production Network [MLPCN]
NIH Chemical Genomics Center [NCGC]

MLPCN Grant: R03 DA026211-01
Assay Provider: Dan Littman, New York University

Assay Overview:

The retinoic acid-related orphan receptor (ROR) gamma is a transcription factor that has a central role in the differentiation of Th17 cells, a subset of T helper cells that secrete the inflammatory cytokines IL-17, IL-17F, and IL-22. Th17 cells have been implicated in graft versus host disease, autoimmune disease and asthma. ROR gamma is induced in naive T helper cells in the presence of TGF-beta combined with IL-6, IL-21, or IL-23, and thereafter directs the expression of the Th17 lineage cytokines. To study the mechanism of action of ROR gamma, an insect cell-based assay was screened. This Drosophila Schneider cell line was stably transfected with two vectors: a gene expressing a fusion of the Gal4 DNA binding domain and ROR gamma transactivation domain under the control of the metallothionine promoter and a Photinus luciferase reporter regulated by the Gal4 binding site enhancer, UAS. The addition of copper to the medium induced expression of the Gal4-ROR gamma fusion that subsequently induced the UAS-luciferase reporter. Small molecule inhibitors of ROR gamma activity are detected by a decrease in luciferase reporter activity.
Protocol
Assay Protocol Summary:
For screening, 600 cells in 4 uL/well were dispensed into white solid 1536-well plates (Grenier) using a solenoid-based dispenser. Following transfer of 23 nL compound or DMSO vehicle by a pin tool, the plates were incubated 1 hr at ambient temperature, and 1 uL/well copper sulfate (700 uM final concentration) was added. The plates were centrifuged 15 s at 1000 RPM and incubated 20 hr at ambient temperature. After addition of 1.5 uL Photinus luciferase detection reagent, the plates were incubated 10 min at ambient temperature and then read by an ViewLux (Perkin Elmer) to detect luminescence.
Keywords: NIH Roadmap, MLPCN, MLI, MLSMR, qHTS, NCGC, qHTS, nuclear hormone receptor, retinoic acid-related orphan receptor, ROR gamma
Comment
Compound Ranking:
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Categorized Comment - additional comments and annotations
From ChEMBL:
Assay Type: Functional
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
6Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
7Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
8Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
9Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
10Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
11Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
13Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14Activity at 0.0000558040 uM (5.5804e-05μM**)% Activity at given concentration.Float%
15Activity at 0.0001180000 uM (0.000118μM**)% Activity at given concentration.Float%
16Activity at 0.0002737148 uM (0.000273715μM**)% Activity at given concentration.Float%
17Activity at 0.0005899000 uM (0.0005899μM**)% Activity at given concentration.Float%
18Activity at 0.00136 uM (0.00135612μM**)% Activity at given concentration.Float%
19Activity at 0.00297 uM (0.00296707μM**)% Activity at given concentration.Float%
20Activity at 0.00432 uM (0.00432102μM**)% Activity at given concentration.Float%
21Activity at 0.00704 uM (0.00703919μM**)% Activity at given concentration.Float%
22Activity at 0.015 uM (0.01475μM**)% Activity at given concentration.Float%
23Activity at 0.033 uM (0.0329733μM**)% Activity at given concentration.Float%
24Activity at 0.061 uM (0.0608644μM**)% Activity at given concentration.Float%
25Activity at 0.082 uM (0.0822688μM**)% Activity at given concentration.Float%
26Activity at 0.145 uM (0.144755μM**)% Activity at given concentration.Float%
27Activity at 0.365 uM (0.36524μM**)% Activity at given concentration.Float%
28Activity at 0.510 uM (0.5102μM**)% Activity at given concentration.Float%
29Activity at 0.826 uM (0.825868μM**)% Activity at given concentration.Float%
30Activity at 1.906 uM (1.90599μM**)% Activity at given concentration.Float%
31Activity at 2.978 uM (2.97753μM**)% Activity at given concentration.Float%
32Activity at 4.961 uM (4.96137μM**)% Activity at given concentration.Float%
33Activity at 10.04 uM (10.0385μM**)% Activity at given concentration.Float%
34Activity at 16.95 uM (16.9495μM**)% Activity at given concentration.Float%
35Activity at 40.67 uM (40.6712μM**)% Activity at given concentration.Float%
36Activity at 53.61 uM (53.6106μM**)% Activity at given concentration.Float%
37Activity at 96.61 uM (96.6079μM**)% Activity at given concentration.Float%
38Activity at 150.4 uM (150.4μM**)% Activity at given concentration.Float%
39Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: R03 DA026211-01

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
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