Abstract 102: Overcoming MET inhibitor resistance in GBM therapy

Abstract

Abstract Glioblastoma (GBM) is the most common and most lethal primary malignant brain tumor. The receptor tyrosine kinase MET is frequently upregulated or over activated in GBM. Clinically applicable MET inhibitors have been developed and tested in the lab and in clinical trials. However, resistance to single modality anti-MET drugs frequently occurs, rendering these agents ineffective. This study aimed to determine the mechanisms of MET inhibitor resistance in GBM and to use the acquired information to develop novel therapeutic approaches to overcome resistance. We investigated two clinically applicable MET inhibitors: PF-02341066, an ATP-competitive small molecule inhibitor of MET, and MetMab, a monovalent monoclonal antibody that binds to the extracellular domain of the MET receptor. We generated PF-02341066 and MetMab resistant GBM cell lines and primary cells by subjecting them to increasing concentrations of drug over a period of time. We utilized RNA sequencing (RNA-seq) and reverse phase protein arrays (RPPA) in addition to death and proliferation assays to identify the pathways altered in the resistant GBM cells compared to wild type GBM cells. We discovered many critical proteins that were altered in the resistant cells lines compared to wild type cells. These included FAK, COX-2, p-FGFR1, Vimentin, mTOR and p-STAT3. There was substantial but not complete overlap between the molecules that were altered in cells resistant to the small molecule as compared to molecules that were changed in cells resistant to the antibody. The protein changes in resistant GBM cells were verified by western blotting. Notably, we discovered that both COX-2 and p-FGFR1 were upregulated in GBM resistant cells and thus investigated whether inhibition of these targets could restore MET inhibitor sensitivity. Celecoxib, an FDA-approved drug, acts to inhibit COX-2 and Debio-1347 acts to inhibit the FGFR family, with a higher affinity for FGFR1. Combining Celecoxib or Debio-1347 with PF-02341066 or MetMab led to increased cell death in resistant cells compared to either agent alone. In addition, the drug combination decreased resistance to cell proliferation inhibition compared to either agent alone, indicating restored sensitivity to both MET inhibitors. We are currently undertaking in vivo experiments to establish whether the combination of celecoxib and MET inhibitors can restore sensitivity to MET inhibition in resistant cells as effectively in vivo as seen with in vitro experiments. These data indicate that MET inhibitor resistance can be overcome by targeting the resulting upregulated pathways using FDA-approved drugs and that multi-drug combinations may revert resistance during treatment. Citation Format: Nichola A. Cruickshanks, Ying Zhang, Sarah Hatef, Julia Wulfkuhle, Isela Gallagher, Alexander Koeppel, David Schiff, See Phan, Stephen Turner, Emanuel Petricoin, Roger Abounader. Overcoming MET inhibitor resistance in GBM therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 102. doi:10.1158/1538-7445.AM2017-102