Role of Endocytosis Proteins in Gefitinib-Mediated EGFR Internalisation in Glioma Cells.

Elisabete Cruz Da Silva,Philippe Rondé,Maxime Lehmann,Stéphane Dedieu,Jessica Thevenard-Devy,Christophe Schneider,Philippe Carl,Laurence Choulier

doi:10.3390/cells10113258

Abstract

EGFR (epidermal growth factor receptor), a member of the ErbB tyrosine kinase receptor family, is a clinical therapeutic target in numerous solid tumours. EGFR overexpression in glioblastoma (GBM) drives cell invasion and tumour progression. However, clinical trials were disappointing, and a molecular basis to explain these poor results is still missing. EGFR endocytosis and membrane trafficking, which tightly regulate EGFR oncosignaling, are often dysregulated in glioma. In a previous work, we showed that EGFR tyrosine kinase inhibitors, such as gefitinib, lead to enhanced EGFR endocytosis into fused early endosomes. Here, using pharmacological inhibitors, siRNA-mediated silencing, or expression of mutant proteins, we showed that dynamin 2 (DNM2), the small GTPase Rab5 and the endocytosis receptor LDL receptor-related protein 1 (LRP-1), contribute significantly to gefitinib-mediated EGFR endocytosis in glioma cells. Importantly, we showed that inhibition of DNM2 or LRP-1 also decreased glioma cell responsiveness to gefitinib during cell evasion from tumour spheroids. By highlighting the contribution of endocytosis proteins in the activity of gefitinib on glioma cells, this study suggests that endocytosis and membrane trafficking might be an attractive therapeutic target to improve GBM treatment.

Highlights

We have previously shown that in, U87, T98G and LN443 GBM cells that express wild type EGFR [23], cytostatic concentrations of gefitinib lead to the accumulation of EGFR in enlarged early endosomes and increase EGF endocytosis, a phenomenon we called gefitinib-mediated endocytosis (GME) [22]
In various GBM cells, gefitinib perturbs membrane trafficking and increasesshowed
GBM cells, we identified threethat endocytosis proteins, dynamin 2 (DNM2), Rab5effiand LRPsiRNA-mediated approaches, we showed

Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. EGFR (epidermal growth factor receptor), a member of the ErbB tyrosine kinase receptor family, is commonly found amplified and/or mutated in near 60% of glioblastoma (GBM), the most aggressive brain tumour. PI3K/Akt (phosphatidyl-inositol-Kinase/Akt), MAPK/ERK (mitogen-activated protein kinases/extracellular signal-regulated kinases), signal transducer and activator of transcription 3 (STAT3), and phospholipase C gamma signalling cascades. These EGFR transduced signals promote GBM cell proliferation and invasion, and tumour progression [1,2]

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