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Published on: September 19, 2018
Phosphoproteomic Analysis of Cell-Based Resistance to BRAF Inhibitor Therapy in Melanoma
Robert Parker1, Laura J Vella2, Dylan Xavier1
1Australian Proteome Analysis Facility, Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, NSW , Australia.
Abstract:
The treatment of melanoma by targeted inhibition of the mutated kinase BRAF with small molecules only temporarily suppresses metastatic disease. In the face of chemical inhibition tumor plasticity, both innate and adaptive, promotes survival through the biochemical and genetic reconfiguration of cellular pathways that can engage proliferative and migratory systems. To investigate this process, high-resolution mass spectrometry was used to characterize the phosphoproteome of this transition in vitro. A simple and accurate, label-free quantitative method was used to localize and quantitate thousands of phosphorylation events. We also correlated changes in the phosphoproteome with the proteome to more accurately determine changes in the activity of regulatory kinases determined by kinase landscape profiling. The abundance of phosphopeptides with sites that function in cytoskeletal regulation, GTP/GDP exchange, protein kinase C, IGF signaling, and melanosome maturation were highly divergent after transition to a drug resistant phenotype.
Insights
Targeted BRAF inhibition in melanoma initially works but drug resistance emerges. Tumor cells adapt by reconfiguring pathways, leading to sustained proliferation and migration, highlighting key molecular changes.
Area of Science:
- Oncology
- Biochemistry
- Molecular Biology
Background:
- Targeted therapy with BRAF inhibitors offers temporary control of metastatic melanoma.
- Tumor cells develop resistance through plasticity, involving biochemical and genetic pathway reconfiguration.
- This adaptive response enables cell survival via proliferative and migratory systems.
Purpose of the Study:
- To investigate the phosphoproteome changes during melanoma drug resistance.
- To characterize the molecular mechanisms underlying tumor plasticity and survival.
- To correlate phosphoproteome and proteome shifts to understand kinase activity.
Main Methods:
- High-resolution mass spectrometry was employed to analyze the phosphoproteome.
- A label-free quantitative method was used to identify and quantify phosphorylation events.
- Kinase landscape profiling correlated phosphoproteome and proteome changes.
Main Results:
- Thousands of phosphorylation events were localized and quantified.
- Significant divergence in phosphopeptides related to cytoskeletal regulation, GTP/GDP exchange, protein kinase C, IGF signaling, and melanosome maturation was observed.
- These changes occurred after the transition to a drug-resistant phenotype.
Conclusions:
- Melanoma drug resistance involves complex phosphoproteome and proteome alterations.
- Understanding these adaptive pathways is crucial for developing more effective melanoma treatments.
- Targeting specific signaling pathways identified in resistant cells may overcome therapeutic limitations.
