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BioAssay: AID 624152

qHTS of GLP-1 Receptor Agonists: Summary

The overall goal of this project was to develop a novel assay for discovering small molecule ligands for class B1 G protein-coupled receptors (GPCRs). Very few agonists (or inverse agonists) with generally weak activity for this entire group of physiologically important receptors are known to date. To establish proof-of-principle that this critical bottleneck can be overcome with a suitable screening approach, our studies focused on the class B1 receptor for glucagon-like peptide-1 (GLP-1R), a potential therapeutic target for diabetes and neurodegenerative disease. ..more
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AID: 624152
Data Source: NCGC (GLPA1000)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2012-05-16
Modify Date: 2012-05-30
Target
Depositor Specified Assays
AIDNameTypeComment
624148qHTS of GLP-1 Receptor Agonists: LOPAC Validationconfirmatory
624172qHTS of GLP-1 Receptor AgonistsconfirmatoryqHTS AID
743262qHTS of GLP-1 Receptor Agonists: Hit Validationconfirmatory
624421qHTS of GLP-1 Receptor Inverse Agonists: Summarysummary
Description:
The overall goal of this project was to develop a novel assay for discovering small molecule ligands for class B1 G protein-coupled receptors (GPCRs). Very few agonists (or inverse agonists) with generally weak activity for this entire group of physiologically important receptors are known to date. To establish proof-of-principle that this critical bottleneck can be overcome with a suitable screening approach, our studies focused on the class B1 receptor for glucagon-like peptide-1 (GLP-1R), a potential therapeutic target for diabetes and neurodegenerative disease.

The assay was developed in a "mix and measure" format using a cell-based functional readout. Toward this goal, stably transfected HEK293 cell clones were established that co-express (i) a cAMP-responsive luciferase reporter gene, and (ii) GLP-1Rs that upon ligand interaction either stimulate (agonists) or inhibit (inverse agonists) luciferase activity. In different cell lines to be used for screening, expressed GLP-1Rs are specifically optimized for high sensitivity detection of either weak agonists (hypersensitive receptor) or inverse agonists (constitutively active receptor mutant). In addition, control cell lines were developed that express either the cAMP-responsive reporter gene without recombinant GLP-1Rs (enabling initial exclusion of GLP-1R independent effects of candidate probes), or the reporter gene together with wild type GLP-1Rs (for confirmation of probe activity at this receptor), or the reporter gene in combination with the constitutively active orphan receptor GPR119 (enabling initial exclusion of GLP-1R independent inhibitory effects of candidate inverse agonists).

To that end, this assay was optimized for 1536-well format and screened against the MLSMR.

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

Grant: NS064851
PI Name: Martin Beinborn, Tufts Medical Center
Protocol
Please see linked AIDs for detailed protocols.
Comment
This project is on-going and will be updated at a later point.
Additional Information
Grant Number: NS064851

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