Bookmark and Share
BioAssay: AID 720652

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

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
_
   
 Tested Compounds
 Tested Compounds
All(1335)
 
 
Active(58)
 
 
Inactive(1112)
 
 
Inconclusive(175)
 
 
 Tested Substances
 Tested Substances
All(1408)
 
 
Active(60)
 
 
Inactive(1166)
 
 
Inconclusive(182)
 
 
 Related BioAssays
 Related BioAssays
AID: 720652
Data Source: NCGC (ROR432)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
Deposit Date: 2013-10-17

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 58
Related Experiments
AIDNameTypeComment
651802qHTS assay for small molecule antagonists of the retinoid-related orphan receptor gamma (ROR-gamma) signaling pathwayConfirmatorydepositor-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. The cytotoxicity of the compounds against ROR response-Luc reporter gene cell line was quantified by measuring the intracellular ATP content.
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 incubation at 37 C and 5% CO2 for 5 hr, 23 nL compound or DMSO vehicle were transferred by a pin tool, then the plates were treated with 1 ul doxycycline to induce reporter gene activity. After 16 hr incubation at 37 C and 5% CO2, 5 uL/well of CellTiter-Glo reagent (Promega) was then added to each plate using a Flying Reagent Dispenser and incubated at room temperature for 30 min. The luminescence intensity was quantified on a ViewLux plate reader (PerkinElmer).
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
Result Definitions
Show more
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Phenotype-Replicate_1Indicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency-Replicate_1*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
3Efficacy-Replicate_1Maximal 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 Comment-Replicate_1Annotation/notes on a particular compound's data or its analysis.String
5Activity_Score-Replicate_1Activity score.Integer
6Curve_Description-Replicate_1A 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_LogAC50-Replicate_1The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlope-Replicate_1The Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2-Replicate_1R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivity-Replicate_1The asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivity-Replicate_1Efficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClass-Replicate_1Numerical encoding of curve description for the fitted Hill equation.Float
13Excluded_Points-Replicate_1Which 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_Response-Replicate_1Maximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.0005899000 uM-Replicate_1 (0.0005899μM**)% Activity at given concentration.Float%
16Activity at 0.00295 uM-Replicate_1 (0.002949μM**)% Activity at given concentration.Float%
17Activity at 0.015 uM-Replicate_1 (0.01475μM**)% Activity at given concentration.Float%
18Activity at 0.033 uM-Replicate_1 (0.03297μM**)% Activity at given concentration.Float%
19Activity at 0.074 uM-Replicate_1 (0.07373μM**)% Activity at given concentration.Float%
20Activity at 0.165 uM-Replicate_1 (0.1649μM**)% Activity at given concentration.Float%
21Activity at 0.369 uM-Replicate_1 (0.3687μM**)% Activity at given concentration.Float%
22Activity at 0.824 uM-Replicate_1 (0.8244μM**)% Activity at given concentration.Float%
23Activity at 1.843 uM-Replicate_1 (1.843μM**)% Activity at given concentration.Float%
24Activity at 4.122 uM-Replicate_1 (4.122μM**)% Activity at given concentration.Float%
25Activity at 9.217 uM-Replicate_1 (9.217μM**)% Activity at given concentration.Float%
26Activity at 20.61 uM-Replicate_1 (20.61μM**)% Activity at given concentration.Float%
27Activity at 46.08 uM-Replicate_1 (46.08μM**)% Activity at given concentration.Float%
28Activity at 92.17 uM-Replicate_1 (92.17μM**)% Activity at given concentration.Float%
29Compound QC-Replicate_1NCGC 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
PageFrom: