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BioAssay: AID 504836

Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in human glioma: Validation

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(1260)
 
 
Active(1)
 
 
Inactive(1240)
 
 
Inconclusive(19)
 
 
 Tested Substances
 Tested Substances
All(1274)
 
 
Active(1)
 
 
Inactive(1254)
 
 
Inconclusive(19)
 
 
AID: 504836
Data Source: NCGC (ERSR000)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2011-06-22
Modify Date: 2011-11-19

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compound: 1
Related Experiments
AIDNameTypeComment
504849Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in human glioma: SummarySummarydepositor-specified cross reference
602332qHTS for Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in Human Glioma: qHTSConfirmatorysame project related to Summary assay
624342qHTS for Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in Human Glioma: Hit ValidationOthersame project related to Summary assay
624348qHTS for Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in Human Glioma: Hit Validation in Cytotoxic CounterscreenOthersame project related to Summary assay
624498Hit Validation Western Blot Assay for Inducers of ERSR: Assessment of GRP78 UpregulationOthersame project related to Summary assay
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 activators are ranked higher than compounds that showed apparent inhibition.
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 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
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 0.00245 uM (0.00245μM**)% Activity at given concentration.Float%
15Activity at 0.012 uM (0.0123μM**)% Activity at given concentration.Float%
16Activity at 0.061 uM (0.0613μM**)% Activity at given concentration.Float%
17Activity at 0.307 uM (0.307μM**)% Activity at given concentration.Float%
18Activity at 1.530 uM (1.53μM**)% Activity at given concentration.Float%
19Activity at 7.660 uM (7.66μM**)% Activity at given concentration.Float%
20Activity at 38.30 uM (38.3μM**)% Activity at given concentration.Float%
21Compound 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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