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EGFR signaling and pharmacology in oncology revealed with innovative BRET-based biosensors
Florence Gross1, Arturo Mancini1, Billy Breton2
1Domain Therapeutics North America Inc., 7171 Frederick-Banting, Saint-Laurent, Quebec, H4S 1Z9, Canada.
Abstract:
Mutations of receptor tyrosine kinases (RTKs) are associated with the development of many cancers by modifying receptor signaling and contributing to drug resistance in clinical settings. We present enhanced bystander bioluminescence resonance energy transfer-based biosensors providing new insights into RTK biology and pharmacology critical for the development of more effective RTK-targeting drugs. Distinct SH2-specific effector biosensors allow for real-time and spatiotemporal monitoring of signal transduction pathways engaged upon RTK activation. Using EGFR as a model, we demonstrate the capacity of these biosensors to differentiate unique signaling signatures, with EGF and Epiregulin ligands displaying differences in efficacy, potency, and responses within different cellular compartments. We further demonstrate that EGFR single point mutations found in Glioblastoma or non-small cell lung cancer, impact the constitutive activity of EGFR and response to tyrosine kinase inhibitor. The BRET-based biosensors are compatible with microscopy, and more importantly characterize the next generation of therapeutics directed against RTKs.
Insights
New biosensors track receptor tyrosine kinase (RTK) signaling in real-time, offering insights into cancer drug resistance and guiding the development of targeted therapies.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Receptor tyrosine kinase (RTK) mutations drive cancer development and drug resistance.
- Understanding RTK signaling is crucial for effective cancer drug development.
Purpose of the Study:
- To develop and validate enhanced bioluminescence resonance energy transfer (BRET)-based biosensors for monitoring RTK signaling.
- To provide new insights into RTK biology and pharmacology for improved targeted therapies.
Main Methods:
- Development of SH2-specific effector biosensors for real-time, spatiotemporal monitoring of RTK signal transduction.
- Utilizing epidermal growth factor receptor (EGFR) as a model system.
- Employing microscopy for BRET-based biosensor compatibility.
Main Results:
- Biosensors successfully differentiated unique signaling signatures of EGFR activation by different ligands (EGF, Epiregulin).
- EGFR mutations associated with glioblastoma and non-small cell lung cancer were shown to affect constitutive activity and drug response.
- Demonstrated differences in ligand efficacy, potency, and cellular compartment responses.
Conclusions:
- Enhanced BRET biosensors offer novel tools for studying RTK signaling dynamics.
- These biosensors are critical for characterizing next-generation therapeutics targeting RTKs.
- The findings facilitate the development of more effective RTK-targeting drugs.
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