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BioAssay: AID 602332

qHTS for Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in Human Glioma: qHTS

Primary protein unfolding takes place for many diseases, including diabetes and hypoxia, , and triggers a reaction in the cells called ER stress response (ERSR). ). ERSR is designed as a repair mechanism, but ultimately leads to cell death via apoptosis if conditions triggering protein unfolding persist (1). The molecular clue to ERSR activation is the enhancement of the expression of key molecular chaperones, including the glucose regulated protein (GRP) 78. GRP78 is the main sensor for protein folding and the main actuator of the ERSR. ..more
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 Tested Compounds
 Tested Compounds
All(415850)
 
 
Active(77)
 
 
Inactive(413250)
 
 
Inconclusive(2652)
 
 
 Tested Substances
 Tested Substances
All(424929)
 
 
Active(77)
 
 
Inactive(422161)
 
 
Inconclusive(2691)
 
 
AID: 602332
Data Source: NCGC (ERSR100)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2012-03-08
Modify Date: 2012-03-13

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 77
Depositor Specified Assays
AIDNameTypeComment
504849Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in human glioma: SummarysummarySummary AID
Description:
Primary protein unfolding takes place for many diseases, including diabetes and hypoxia, , and triggers a reaction in the cells called ER stress response (ERSR). ). ERSR is designed as a repair mechanism, but ultimately leads to cell death via apoptosis if conditions triggering protein unfolding persist (1). The molecular clue to ERSR activation is the enhancement of the expression of key molecular chaperones, including the glucose regulated protein (GRP) 78. GRP78 is the main sensor for protein folding and the main actuator of the ERSR.

In a collaboration between the Southern Research Institute and the NIH Chemical Genomics Center, a cell based screen was developed that reliably uses GRP78 as a sensor to identify small molecules as inducers of ERSR. In resting conditions, GRP78 binds proteins and acts as a natural repressor. Once unfolded proteins are present in the ER, GRP78 releases some of the repressors and binds to the unfolded proteins. The release of the repressors executes the ERSR program. Using a human glioma cell line in a miniaturized, luminescent format, a high-throughput screen was developed to identify actuators of the ERSR.

(1) Soboloff, J. and S. A. Berger (2002). "Sustained ER Ca2+ depletion suppresses protein synthesis and induces activation-enhanced cell death in mast cells." J Biol Chem 277(16): 13812-20.

NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]

MLPCN Grant: DA031669-01
Assay Submitter (PI): Maurizio Grimaldi, Southern Research Institute
Protocol
Stable transfected human glioma cell line, U87-MG, are dispensed at density 1000 cells/well into Greiner white solid bottom 1536-well assay plates. Compounds are transferred via Kalypsys pin tool equipped with 1536-pin array (10nl slotted pins, V&P Scientific, San Diego, CA). The assay plates will be incubated for 16 hours and developed by adding appropriate quantities of the Promega Bright Glo reagent. The plates will be incubated at room temperature for 10 minutes prior to being transferred into a ViewLux high-throughput CCD imager (PerkinElmer), wherein end-point measurements of luminescence are acquired.
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
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 0.00245 uM (0.00245μM**)% Activity at given concentration.Float%
16Activity at 0.00549 uM (0.00549μM**)% Activity at given concentration.Float%
17Activity at 0.012 uM (0.0123μM**)% Activity at given concentration.Float%
18Activity at 0.027 uM (0.0274μM**)% Activity at given concentration.Float%
19Activity at 0.061 uM (0.0613μM**)% Activity at given concentration.Float%
20Activity at 0.137 uM (0.137μM**)% Activity at given concentration.Float%
21Activity at 0.282 uM (0.28152μM**)% Activity at given concentration.Float%
22Activity at 0.366 uM (0.365636μM**)% Activity at given concentration.Float%
23Activity at 0.561 uM (0.561236μM**)% Activity at given concentration.Float%
24Activity at 0.830 uM (0.829567μM**)% Activity at given concentration.Float%
25Activity at 1.581 uM (1.58063μM**)% Activity at given concentration.Float%
26Activity at 2.411 uM (2.4112μM**)% Activity at given concentration.Float%
27Activity at 3.612 uM (3.61163μM**)% Activity at given concentration.Float%
28Activity at 7.587 uM (7.58671μM**)% Activity at given concentration.Float%
29Activity at 10.35 uM (10.346μM**)% Activity at given concentration.Float%
30Activity at 16.10 uM (16.1032μM**)% Activity at given concentration.Float%
31Activity at 32.69 uM (32.6884μM**)% Activity at given concentration.Float%
32Activity at 41.28 uM (41.2754μM**)% Activity at given concentration.Float%
33Activity at 75.75 uM (75.746μM**)% Activity at given concentration.Float%
34Activity at 111.4 uM (111.388μM**)% Activity at given concentration.Float%
35Activity at 167.2 uM (167.179μM**)% Activity at given concentration.Float%
36Activity at 290.9 uM (290.854μM**)% Activity at given concentration.Float%
37Activity at 384.0 uM (384μM**)% Activity at given concentration.Float%
38Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: DA031669

Data Table (Concise)
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