AID	Name	Type	Comment
504849	Inducers of the Endoplasmic Reticulum Stress Response (ERSR) in human glioma: Summary	summary

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

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.

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.

Grant Number: DA031669