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BioAssay: AID 651802

qHTS assay for small molecule antagonists of the retinoid-related orphan receptor gamma (ROR-gamma) signaling pathway

Nuclear steroid hormone receptors are involved in many biological processes including inflammatory responses, cell proliferation, and apoptosis. Environmental factors or compounds disrupting these nuclear receptor pathways and gene expression may promote susceptibility to immunotoxicants and autoimmune diseases. One important group of these expression factors are the retinoid-related orphan receptors. To assess compounds that may interfere with the retinoid-related orphan receptor gamma (ROR-gamma) transcription factor, a cell-based luciferase assay was used. ..more
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 Tested Compounds
 Tested Compounds
All(1335)
 
 
Active(143)
 
 
Inactive(1050)
 
 
Inconclusive(152)
 
 
 Tested Substances
 Tested Substances
All(1408)
 
 
Active(151)
 
 
Inactive(1102)
 
 
Inconclusive(155)
 
 
 Related BioAssays
 Related BioAssays
AID: 651802
Data Source: NCGC (ROR993)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
Deposit Date: 2012-11-20

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 143
Related Experiments
AIDNameTypeComment
720652qHTS assay for small molecule antagonists of the retinoid-related orphan receptor gamma (ROR-gamma) signaling pathway - cell viability counter screenConfirmatorydepositor-specified cross reference
Description:
National Center for Advancing Translational Sciences [NCATS]
NIH Chemical Genomics Center [NCGC]
National Institutes of Environmental Health Sciences [NIEHS]
National Toxicology Program [NTP]

Assay Overview:

Nuclear steroid hormone receptors are involved in many biological processes including inflammatory responses, cell proliferation, and apoptosis. Environmental factors or compounds disrupting these nuclear receptor pathways and gene expression may promote susceptibility to immunotoxicants and autoimmune diseases. One important group of these expression factors are the retinoid-related orphan receptors. To assess compounds that may interfere with the retinoid-related orphan receptor gamma (ROR-gamma) transcription factor, a cell-based luciferase assay was used.
The CHO cell-based assay utilizes a TET-inducible ROR expression factor and ROR response element-LUC reporter gene. Reporter activity was induced using doxycycline and measured after 16hr incubation. Luminescence was read using Viewlux (PerkinElmer), where a decrease in reporter activity would show a decrease in luminescence.
Protocol
Assay Protocol Summary:

1,000 cells in 4 uL/well were dispensed into white wall/solid bottom 1536-well plates using a Multidrop Combi (Thermo Scientific) dispenser. Following transfer of 23 nL compound or DMSO vehicle by a pin tool, the plates were treated with 1 ul doxycycline to induce reporter gene activity. After 16hr incubation at 37C and 5% CO2, 5uL one-glo (promega) was added to each plate and following 30 min incubation at room temperature, luminescence was quantified on a ViewLux (PerkinElmer) plate reader.
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 PubChem:
Assay Cell Type: CHO
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
5Activity_ScoreActivity score.Integer
6Curve_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
7Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13Excluded_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
14Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.0005899000 uM (0.0005899μM**)% Activity at given concentration.Float%
16Activity at 0.00295 uM (0.002949μM**)% Activity at given concentration.Float%
17Activity at 0.015 uM (0.01475μM**)% Activity at given concentration.Float%
18Activity at 0.033 uM (0.03297μM**)% Activity at given concentration.Float%
19Activity at 0.074 uM (0.07373μM**)% Activity at given concentration.Float%
20Activity at 0.165 uM (0.1649μM**)% Activity at given concentration.Float%
21Activity at 0.369 uM (0.3687μM**)% Activity at given concentration.Float%
22Activity at 0.824 uM (0.8244μM**)% Activity at given concentration.Float%
23Activity at 1.843 uM (1.843μM**)% Activity at given concentration.Float%
24Activity at 4.122 uM (4.122μM**)% Activity at given concentration.Float%
25Activity at 9.217 uM (9.217μM**)% Activity at given concentration.Float%
26Activity at 20.61 uM (20.61μM**)% Activity at given concentration.Float%
27Activity at 46.08 uM (46.08μM**)% Activity at given concentration.Float%
28Activity at 92.17 uM (92.17μM**)% Activity at given concentration.Float%
29Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.

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