The role of EGFR double minutes in modulating the response of malignant gliomas to radiotherapy

// Yi-Hong Zhou 1 , Yumay Chen 2 , Yuanjie Hu 1 , Liping Yu 3 , Katherine Tran 3 , Erich Giedzinski 3 , Ning Ru 1 , Alex Gau 1 , Francine Pan 1 , Jiao Qiao 1 , Naomi Atkin 1 , Khang Chi Ly 1 , Nathan Lee 1 , Eric R. Siegel 4 , Mark E. Linskey 1 , Ping Wang 2 and Charles Limoli 3 1 UC Irvine Brain Tumor Laboratory and Department of Surgery, University of California Irvine, Irvine, CA, USA 2 UC Irvine Diabetes Center and Department of Medicine, University of California Irvine, Irvine, CA, USA 3 Department of Radiation Oncology, University of California Irvine, Irvine, CA, USA 4 Departments of Biostatistics, University of Arkansas for Medical Sciences, Little Rock, AR, USA Correspondence to: Yi-Hong Zhou, email: yihongz@uci.edu Charles Limoli, email: climoli@uci.edu Keywords: glioblastoma multiforme, EGFR, double minute (DM) chromosome mis-segregation, radiation, tumor heterogeneity Received: March 20, 2017      Accepted: August 04, 2017      Published: September 08, 2017 ABSTRACT EGFR amplification in cells having double minute chromosomes (DM) is commonly found in glioblastoma multiforme (GBM); however, how much it contributes to the current failure to treat GBM successfully is unknown. We studied two syngeneic primary cultures derived from a GBM with and without cells carrying DM, for their differential molecular and metabolic profiles, in vivo growth patterns, and responses to irradiation (IR). Each cell line has a distinct molecular profile consistent with an invasive “go” (with DM) or angiogenic “grow” phenotype (without DM) demonstrated in vitro and in intracranial xenograft models. Cells with DM were relatively radio-resistant and used higher glycolytic respiration and lower oxidative phosphorylation in comparison to cells without them. The DM-containing cell was able to restore tumor heterogeneity by mis-segregation of the DM-chromosomes, giving rise to cell subpopulations without them. As a response to IR, DM-containing cells switched their respiration from glycolic metabolism to oxidative phosphorylation and shifted molecular profiles towards that of cells without DM. Irradiated cells with DM showed the capacity to alter their extracellular microenvironment to not only promote invasiveness of the surrounding cells, regardless of DM status, but also to create a pro-angiogenic tumor microenvironment. IR of cells without DM was found primarily to increase extracellular MMP2 activity. Overall, our data suggest that the DM-containing cells of GBM are responsible for tumor recurrence due to their high invasiveness and radio-resistance and the mis-segregation of their DM chromosomes, to give rise to fast-growing cells lacking DM chromosomes.