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

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, hits identified in the primary ERSR screen were characterized for their effects on wild type GRP78 upregulation using a western blot secondary assay.

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.

The cells will be plated and exposed to the compounds for variable times (4, 8, 24h) to determine the time profile of the effect. Cells will be harvested and the cell extracts will be subjected to SDS-PAGE and Western blotting procedures to evaluate the extent of upregulation of GRP78.

If band is observed, compounds are considered "active" and assigned a score of 90. If not band is observed, compounds are considered "inactive" and assigned a score of 10.

Grant Number: DA031669