Integrated Cell-Based Platform to Study EGFR Activation and Transactivation
| Autor: | Stephane Parent, Nathalie Rouleau, Marie-Elaine Caruso, Paule Clément, Vincent Dupriez, Roger Bossé |
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| Rok vydání: | 2013 |
| Předmět: |
Drug Evaluation
Preclinical Cell Count Biosensing Techniques CHO Cells Biology Pharmacology Transactivation chemistry.chemical_compound Cricetulus Cell surface receptor Cricetinae Drug Discovery medicine Animals Cyclic adenosine monophosphate Epidermal growth factor receptor Receptor G protein-coupled receptor Equipment Design Flow Cytometry High-Throughput Screening Assays Equipment Failure Analysis ErbB Receptors Systems Integration Mechanism of action chemistry Cancer research biology.protein Molecular Medicine Biological Assay Signal transduction medicine.symptom |
| Zdroj: | ASSAY and Drug Development Technologies. 11:423-434 |
| ISSN: | 1557-8127 1540-658X |
| Popis: | The epidermal growth factor receptor (EGFR) pathway is one of the most deregulated molecular pathways in human epithelial cancers. Many approved drugs were optimized to directly target EGFR but yielded only modest clinical improvement in cancer patients due to low efficacy and drug resistance. Transactivation of EGFR by other cell surface receptors such as G-protein-coupled receptors (GPCRs) was proposed to explain this lack of efficacy. Even if direct EGFR activation and transactivation by GPCR contribute to the activation of the same signaling pathways, they are often studied as independent events resulting in partial investigation of a drug's mechanism of action. We present a novel high-throughput approach that integrates interrogation of direct activation of EGFR and its transactivation via GPCR activation. Using distinct technology platforms, three readouts were used to measure (1) direct activation of GPCR via cyclic adenosine monophosphate (cAMP) detection, (2) direct activation of EGFR through the release of intracellular Ca(2+), and (3) EGFR transactivation by GPCR using the detection of p-extracellular-signal-regulated kinases 1/2 (p-ERK1/2). In addition to being simple, quick, and homogenous, our methods were shown to be more sensitive than those in current use. These enabling tools should improve the knowledge pertaining to GPCRs and receptor tyrosine kinases trans-regulation and facilitate the design of more potent and better targeted new therapeutic strategies. |
| Databáze: | OpenAIRE |
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