Bookmark and Share
BioAssay: AID 504467

qHTS screen for small molecules that inhibit ELG1-dependent DNA repair in human embryonic kidney (HEK293T) cells expressing luciferase-tagged ELG1

Cancer cells divide rapidly compared to normal cells in the body. During cell division, cancer cells need to duplicate their genome by DNA replication. The failure of genome duplication leads to cancer cells' death. Therefore, inhibitors of DNA replication could specifically kill cancer cells. Based on this concept, many chemotherapeutic agents were developed and have been used to treat cancer more ..
_
   
 Tested Compounds
 Tested Compounds
All(330113)
 
 
Active(7652)
 
 
Inactive(235626)
 
 
Inconclusive(86871)
 
 
 Tested Substances
 Tested Substances
All(330279)
 
 
Active(7656)
 
 
Inactive(235723)
 
 
Inconclusive(86900)
 
 
AID: 504467
Data Source: NCGC (ELG233)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-03-16

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 7652
Depositor Specified Assays
AIDNameTypeComment
493125qHTS assay for small molecules that inhibit ELG1-dependent DNA repair in human embryonic kidney (HEK293T) cells expressing luciferase-tagged ELG1: Summarysummary
493107Validation screen for small molecules that inhibit ELG1-dependent DNA repair in human embryonic kidney (HEK293T) cells expressing luciferase-tagged ELG1confirmatory
Description:
NIH Molecular Libraries Probe Production Network [MLPCN]
NIH Chemical Genomics Center [NCGC]

MLPCN Grant: 1 R03 MH092164-01
Assay Provider: Kyungjae Myung, NHGRI, NIH

Keywords: MLPCN, qHTS, NCGC, genotoxicity, cancer

Assay Overview:

Cancer cells divide rapidly compared to normal cells in the body. During cell division, cancer cells need to duplicate their genome by DNA replication. The failure of genome duplication leads to cancer cells' death. Therefore, inhibitors of DNA replication could specifically kill cancer cells. Based on this concept, many chemotherapeutic agents were developed and have been used to treat cancer patients. However, there are a limited number of agents available in the clinical setting, and some of those that do exist have low efficacy and severe side effects. Therefore, if we can find the compounds that effectively block DNA replication either by directly damaging DNA or by inhibiting other cellular mechanisms, the options for cancer treatment will be broadened. With this in mind, we developed a quantitative high-throughput ELG1-luciferase reporter gene assay. Small molecules that specifically block ELG1 function in response to DNA damage can be identified by monitoring the suppression of luciferase activity. Since ELG1 defines a pathway for repairing DNA damage, agents blocking ELG1 function can be used to sensitize cancer cells to chemotherapeutic agents whose mechanism of action involves the induction of DNA damage.
Protocol
Assay Protocol Summary:

For genotoxicity screening, 2,000 cells in 4 uL/well were dispensed into white, solid-bottom 1536-well plates using a solenoid-based dispenser. Following transfer of 1 uL 5-Fluorouridine and 23 nL compound or DMSO vehicle by a pin tool, the plates were incubated 16 hr at 37oC and 5% CO2. 5 ul ONE-Glo (Promega) was then added to each plate and following a 30 min incubation at room temperature, luciferase activity was quantified on a ViewLux CCD-based 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
Show more
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 0.0005897200 uM (0.00058972μM**)% Activity at given concentration.Float%
15Activity at 0.00132 uM (0.00131825μM**)% Activity at given concentration.Float%
16Activity at 0.00295 uM (0.00294939μM**)% Activity at given concentration.Float%
17Activity at 0.00659 uM (0.00659275μM**)% Activity at given concentration.Float%
18Activity at 0.015 uM (0.0147467μM**)% Activity at given concentration.Float%
19Activity at 0.033 uM (0.0329625μM**)% Activity at given concentration.Float%
20Activity at 0.074 uM (0.0737375μM**)% Activity at given concentration.Float%
21Activity at 0.165 uM (0.164825μM**)% Activity at given concentration.Float%
22Activity at 0.369 uM (0.368698μM**)% Activity at given concentration.Float%
23Activity at 0.824 uM (0.8241μM**)% Activity at given concentration.Float%
24Activity at 1.843 uM (1.84339μM**)% Activity at given concentration.Float%
25Activity at 4.120 uM (4.1205μM**)% Activity at given concentration.Float%
26Activity at 9.217 uM (9.21694μM**)% Activity at given concentration.Float%
27Activity at 20.60 uM (20.6025μM**)% Activity at given concentration.Float%
28Activity at 46.08 uM (46.08μM**)% Activity at given concentration.Float%
29Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: 1 R03 MH092164-01

Data Table (Concise)
Classification
PageFrom: