Supplementary Table 2 from Vemurafenib Resistance Signature by Proteome Analysis Offers New Strategies and Rational Therapeutic Concepts

Paulitschke, Verena; Berger, Walter; Paulitschke, Philipp; Hofstätter, Elisabeth; Knapp, Bernhard; Dingelmaier-Hovorka, Ruth; Födinger, Dagmar; Jäger, Walter; Szekeres, Thomas; Meshcheryakova, Anastasia; Bileck, Andrea; Pirker, Christine; Pehamberger, Hubert; Gerner, Christopher; Kunstfeld, Rainer

doi:10.1158/1535-7163.22502334.v1

15357163mct140701-sup-136538_1_supp_2795901_ngzvvv.docx (19.42 kB)

Supplementary Table 2 from Vemurafenib Resistance Signature by Proteome Analysis Offers New Strategies and Rational Therapeutic Concepts

journal contribution

posted on 2023-04-03, 14:26 authored by Verena Paulitschke, Walter Berger, Philipp Paulitschke, Elisabeth Hofstätter, Bernhard Knapp, Ruth Dingelmaier-Hovorka, Dagmar Födinger, Walter Jäger, Thomas Szekeres, Anastasia Meshcheryakova, Andrea Bileck, Christine Pirker, Hubert Pehamberger, Christopher Gerner, Rainer Kunstfeld

Comparison of protein expression in A375 and M24met by 2D-gel electrophoresis. Corresponding proteins which are assigned in the 2-D gel in Figure 2. Accession numbers are from the Uniprot database. Vol indicates the volume identified by 2D-gel electrophoresis n.d =not determined

History

ARTICLE 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. Mol Cancer Ther; 14(3); 757–68. ©2015 AACR.