New Mechanisms of Resistance to MEK Inhibitors in Melanoma Revealed by Intravital Imaging

• Published in: Cancer research (Chicago, Ill.) Ill.), 2018-01, Vol.78 (2), p.542-557
• Main Authors: Brighton, Hailey E, Angus, Steven P, Bo, Tao, Roques, Jose, Tagliatela, Alicia C, Darr, David B, Karagoz, Kubra, Sciaky, Noah, Gatza, Michael L, Sharpless, Norman E, Johnson, Gary L, Bear, James E
• Format: Article
• Language: English
• Subjects: 1-Phosphatidylinositol 3-kinase; AKT protein; Animal models; Animals; Apoptosis - drug effects; Biomarkers, Tumor - genetics; c-Kit protein; Cancer; Case-Control Studies; Cell Proliferation - drug effects; Collagen; Drug resistance; Drug Resistance, Neoplasm; Gene Expression Profiling; Humans; Inhibitors; Intravital Microscopy - methods; Longitudinal Studies; MAP kinase; MAP Kinase Kinase 1 - antagonists & inhibitors; MEK inhibitors; Melanoma; Melanoma - drug therapy; Melanoma - metabolism; Melanoma - pathology; Mesenchyme; Metastases; Mice; Mutation; Prognosis; Protein Kinase Inhibitors - pharmacology; Pyridones - pharmacology; Pyrimidinones - pharmacology; Raf protein; Signal Transduction; Stroma; Transcription; Tumor cells; Tumor Cells, Cultured; Tumors
• ISSN: 0008-5472; 1538-7445
• DOI: 10.1158/0008-5472.can-17-1653

Targeted therapeutics that are initially effective in cancer patients nearly invariably engender resistance at some stage, an inherent challenge in the use of any molecular-targeted drug in cancer settings. In this study, we evaluated resistance mechanisms arising in metastatic melanoma to MAPK pathway kinase inhibitors as a strategy to identify candidate strategies to limit risks of resistance. To investigate longitudinal responses, we developed an intravital serial imaging approach that can directly visualize drug response in an inducible RAF-driven, autochthonous murine model of melanoma incorporating a fluorescent reporter allele (tdTomatoLSL). Using this system, we visualized formation and progression of tumors , starting from the single-cell level longitudinally over time. Reliable reporting of the status of primary murine tumors treated with the selective MEK1/2 inhibitor (MEKi) trametinib illustrated a time-course of initial drug response and persistence, followed by the development of drug resistance. We found that tumor cells adjacent to bundled collagen had a preferential persistence in response to MEKi. Unbiased transcriptional and kinome reprogramming analyses from selected treatment time points suggested increased c-Kit and PI3K/AKT pathway activation in resistant tumors, along with enhanced expression of epithelial genes and epithelial-mesenchymal transition downregulation signatures with development of MEKi resistance. Similar trends were observed following simultaneous treatment with BRAF and MEK inhibitors aligned to standard-of-care combination therapy, suggesting these reprogramming events were not specific to MEKi alone. Overall, our results illuminate the integration of tumor-stroma dynamics with tissue plasticity in melanoma progression and provide new insights into the basis for drug response, persistence, and resistance. A longitudinal study tracks the course of MEKi treatment in an autochthonous imageable murine model of melanoma from initial response to therapeutic resistance, offering new insights into the basis for drug response, persistence, and resistance. .