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BioAssay: AID 651754

qHTS assay for small molecule disruptors of the mitochondrial membrane potential - 1 hr assay

Mitochondrial membrane potential (MMP), one of the parameters for mitochondrial function, is generated by mitochondrial electron transport chain that creates an electrochemical gradient by a series of redox reactions. This gradient drives the synthesis of ATP, a crucial molecule for various cellular processes. Measuring MMP in living cells is commonly used to assess the effect of chemicals on more ..
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 Tested Compounds
 Tested Compounds
All(1335)
 
 
Active(126)
 
 
Inactive(1063)
 
 
Inconclusive(150)
 
 
 Tested Substances
 Tested Substances
All(1408)
 
 
Active(131)
 
 
Inactive(1122)
 
 
Inconclusive(155)
 
 
 Related BioAssays
 Related BioAssays
AID: 651754
Data Source: NCGC (MMP193)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
Deposit Date: 2012-11-06

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 126
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:

Mitochondrial membrane potential (MMP), one of the parameters for mitochondrial function, is generated by mitochondrial electron transport chain that creates an electrochemical gradient by a series of redox reactions. This gradient drives the synthesis of ATP, a crucial molecule for various cellular processes. Measuring MMP in living cells is commonly used to assess the effect of chemicals on mitochondrial function. A homogenous cell-based MMP assay using a water soluble mitochondrial membrane potential sensor (Mito-MPS dye developed by BD Biosciences) was used to evaluate chemically induced mitochondrial toxicity. The dye accumulates in the mitochondria of healthy cells as aggregates, with red fluorescence. The mitochondrial potential collapses and the dye can no longer accumulate in the mitochondria and remains in the cytoplasm in monomers, with green fluorescence.
Protocol
Assay Protocol Summary:

2,000 HepG2 cells in 5 uL/well were dispensed into black, clear-bottom 1536-well plates using a Multidrop Combi (Thermo Scientific) dispenser. The assay plates were incubated overnight at 37C to allow cells to attach to the wells, followed by the transfer of 23 nL compound or DMSO vehicle by a pin tool (Kalypsys). The plates were incubated for 1 hr at 37C and 5% CO2, followed by the addition of 5 uL of 2X Mito-MPS dye (BD Biosciences) into each well using Flying Reagent Dispenser (Aurora Discovery) and then following 30 min incubation at 37C. Fluorescence intensity (490nm excitation and 535nm emission for monomers; and 540nm excitation and 590nm emissions for aggregates) was measured using an Envision 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.
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
5W590-Activity_ScoreActivity score.Integer
6W590-Curve_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
7W590-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8W590-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9W590-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10W590-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11W590-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12W590-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13W590-Excluded_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
14W590-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15W590-Activity at 0.0005899000 uM (0.0005899μM**)% Activity at given concentration.Float%
16W590-Activity at 0.00295 uM (0.002949μM**)% Activity at given concentration.Float%
17W590-Activity at 0.015 uM (0.01475μM**)% Activity at given concentration.Float%
18W590-Activity at 0.033 uM (0.03297μM**)% Activity at given concentration.Float%
19W590-Activity at 0.074 uM (0.07373μM**)% Activity at given concentration.Float%
20W590-Activity at 0.165 uM (0.1649μM**)% Activity at given concentration.Float%
21W590-Activity at 0.369 uM (0.3687μM**)% Activity at given concentration.Float%
22W590-Activity at 0.824 uM (0.8244μM**)% Activity at given concentration.Float%
23W590-Activity at 1.843 uM (1.843μM**)% Activity at given concentration.Float%
24W590-Activity at 4.122 uM (4.122μM**)% Activity at given concentration.Float%
25W590-Activity at 9.217 uM (9.217μM**)% Activity at given concentration.Float%
26W590-Activity at 20.61 uM (20.61μM**)% Activity at given concentration.Float%
27W590-Activity at 46.08 uM (46.08μM**)% Activity at given concentration.Float%
28W590-Activity at 92.17 uM (92.17μM**)% Activity at given concentration.Float%
29Ratio-Activity_ScoreActivity score.Integer
30Ratio-Curve_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
31Ratio-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
32Ratio-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
33Ratio-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
34Ratio-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
35Ratio-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
36Ratio-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
37Ratio-Excluded_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
38Ratio-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
39Ratio-Activity at 0.0005899000 uM (0.0005899μM**)% Activity at given concentration.Float%
40Ratio-Activity at 0.00295 uM (0.002949μM**)% Activity at given concentration.Float%
41Ratio-Activity at 0.015 uM (0.01475μM**)% Activity at given concentration.Float%
42Ratio-Activity at 0.033 uM (0.03297μM**)% Activity at given concentration.Float%
43Ratio-Activity at 0.074 uM (0.07373μM**)% Activity at given concentration.Float%
44Ratio-Activity at 0.165 uM (0.1649μM**)% Activity at given concentration.Float%
45Ratio-Activity at 0.369 uM (0.3687μM**)% Activity at given concentration.Float%
46Ratio-Activity at 0.824 uM (0.8244μM**)% Activity at given concentration.Float%
47Ratio-Activity at 1.843 uM (1.843μM**)% Activity at given concentration.Float%
48Ratio-Activity at 4.122 uM (4.122μM**)% Activity at given concentration.Float%
49Ratio-Activity at 9.217 uM (9.217μM**)% Activity at given concentration.Float%
50Ratio-Activity at 20.61 uM (20.61μM**)% Activity at given concentration.Float%
51Ratio-Activity at 46.08 uM (46.08μM**)% Activity at given concentration.Float%
52Ratio-Activity at 92.17 uM (92.17μM**)% Activity at given concentration.Float%
53W535-Activity_ScoreActivity score.Integer
54W535-Curve_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
55W535-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
56W535-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
57W535-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
58W535-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
59W535-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
60W535-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
61W535-Excluded_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
62W535-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
63W535-Activity at 0.0005898618 uM (0.000589862μM**)% Activity at given concentration.Float%
64W535-Activity at 0.00295 uM (0.00294931μM**)% Activity at given concentration.Float%
65W535-Activity at 0.015 uM (0.0147465μM**)% Activity at given concentration.Float%
66W535-Activity at 0.033 uM (0.0329954μM**)% Activity at given concentration.Float%
67W535-Activity at 0.074 uM (0.0737327μM**)% Activity at given concentration.Float%
68W535-Activity at 0.165 uM (0.164977μM**)% Activity at given concentration.Float%
69W535-Activity at 0.369 uM (0.368664μM**)% Activity at given concentration.Float%
70W535-Activity at 0.825 uM (0.824885μM**)% Activity at given concentration.Float%
71W535-Activity at 1.843 uM (1.84332μM**)% Activity at given concentration.Float%
72W535-Activity at 4.120 uM (4.11982μM**)% Activity at given concentration.Float%
73W535-Activity at 9.217 uM (9.21659μM**)% Activity at given concentration.Float%
74W535-Activity at 20.60 uM (20.5991μM**)% Activity at given concentration.Float%
75W535-Activity at 46.08 uM (46.0829μM**)% Activity at given concentration.Float%
76W535-Activity at 92.17 uM (92.1659μM**)% Activity at given concentration.Float%
77Compound 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
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