Vemurafenib Resistance Signature by Proteome Analysis Offers New Strategies and Rational Therapeutic Concepts

Abstract

The FDA-approved BRAF inhibitor vemurafenib achieves outstanding clinical response rates in patients with melanoma, but early resistance is common. Understanding the pathologic mechanisms of drug resistance and identification of effective therapeutic alternatives are key scientific challenges in the melanoma setting. Using proteomic techniques, including shotgun analysis and 2D-gel electrophoresis, we identified a comprehensive signature of the vemurafenib-resistant M24met in comparison with the vemurafenib-sensitive A375 melanoma cell line. The resistant cells were characterized by loss of differentiation, induction of transformation, enhanced expression of the lysosomal compartment, increased potential for metastasis, migration, adherence and Ca2(+) ion binding, enhanced expression of the MAPK pathway and extracellular matrix proteins, and epithelial-mesenchymal transformation. The main features were verified by shotgun analysis with QEXACTIVE orbitrap MS, electron microscopy, lysosomal staining, Western blotting, and adherence assay in a VM-1 melanoma cell line with acquired vemurafenib resistance. On the basis of the resistance profile, we were able to successfully predict that a novel resveratrol-derived COX-2 inhibitor, M8, would be active against the vemurafenib-resistant but not the vemurafenib-sensitive melanoma cells. Using high-throughput methods for cell line and drug characterization may thus offer a new way to identify key features of vemurafenib resistance, facilitating the design of effective rational therapeutic alternatives.

Proteomic identification of a marker signature for MAPK i resistance in melanoma

“…Comparing the sensitive (S1) and resistant (R1) melanoma cells by zymography, we detected an increase in MMP2 (matrix metalloproteinase 2) activity in the resistant cells (R1; Fig D), a feature associated with invasiveness in EMT (Bae et al , ). Consistently, N‐cadherin levels were increased in culture R1 (Fig E), which is both in line with the proteome data and our previous publication (Paulitschke et al , ).…”

“…Previously, we identified in an acquired resistance model seven characteristics that were associated with vemurafenib resistance (Paulitschke et al , ). We were also able to confirm these data in the primary cells tested in this study.…”

Targeting breast cancer-associated fibroblasts to improve anti-cancer therapy

“…It is well established that different kinds of cancer cells derived from one tissue may display very profound differences in genetic aberrations, proteome expression profiles and drug responsiveness [16,17]. While it seems plausible, it was not yet systematically investigated whether different kinds of cancer cell activities may trigger the establishment of different functional subtypes of CAFs.…”

Multi-omics Analysis of Serum Samples Demonstrates Reprogramming of Organ Functions Via Systemic Calcium Mobilization and Platelet Activation in Metastatic Melanoma

“…Using mass spectrometry-based proteomics, we have previously investigated melanoma and neighboring stroma cells, focusing on the identification of biomarkers associated with intrinsic and extrinsic drug resistance (5). In the present article, we followed the question how metastatic melanoma may reprogram distant organ functions, and how these changes may be depicted in serum signatures taken from patients treated in a phase II trial on metastatic melanoma.…”