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BioAssay: AID 624342

qHTS for Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in Human Glioma: Hit 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(3)
 
 
Active(3)
 
 
 Tested Substances
 Tested Substances
All(3)
 
 
Active(3)
 
 
AID: 624342
Data Source: NCGC (ERSR200)
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2012-06-27

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 3
Related Experiments
AIDNameTypeComment
504849Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in human glioma: SummarySummarydepositor-specified cross reference: Summary AID
504836Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in human glioma: ValidationConfirmatorysame project related to Summary assay
602332qHTS for Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in Human Glioma: qHTSConfirmatorysame 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. In this assay, the hits identified from primary screen are re-run using the same protocol.


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

MLPCN Grant: DA031669
Assay Submitter (PI): Maurizio Grimaldi, Southern Research Institute

(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.
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
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1IC50 (uM)FloatμM
2Compound QCString
Additional Information
Grant Number: DA031669

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
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