AKT can modulate the in vitro response of HNSCC cells to irreversible EGFR inhibitors

// Renato Jose Silva-Oliveira 1 , Matias Melendez 1 , Olga Martinho 1, 2, 3 , Maicon F. Zanon 1 , Luciano de Souza Viana 1, 4 , Andre Lopes Carvalho 1 and Rui Manuel Reis 1, 2, 3 1 Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos, Brazil 2 Life and Health Sciences Research Institute (ICVS), Health Sciences School, University of Minho, Braga, Portugal 3 ICVS/3B’s-PT Government Associate Laboratory, Braga, Portugal 4 Department of Medical Oncology, Barretos Cancer Hospital, Barretos, Brazil Correspondence to: Rui Manuel Reis, email: ruireis.hcb@gmail.com Keywords: HNSCC, anti-EGFR, anti- AKT, AKT1, resistance Received: July 20, 2016     Accepted: May 16, 2017     Published: June 07, 2017 ABSTRACT Epidermal growth factor receptor (EGFR) is overexpressed in up to 90% of head and neck squamous cell carcinoma (HNSCC) tumors. Cetuximab is the first targeted (anti-EGFR) therapy approved for the treatment of HNSCC patients. However, its efficacy is limited due to primary and secondary resistance, and there is no predict biomarkers of response. New generation of EGFR inhibitors with pan HER targeting and irreversible action, such as afatinib and allitinib, represents a significant therapeutic promise. In this study, we intend to compare the potential cytotoxicity of two anti-EGFR inhibitors (afatinib and allitinib) with cetuximab and to identify potential predictive biomarkers of response in a panel of HNSCC cell lines. The mutational analysis in the eight HNSCC cell lines revealed an EGFR mutation (p.H773Y) and gene amplification in the HN13 cells. According to the growth inhibition score (GI), allitinib was the most cytotoxic drug, followed by afatinib and finally cetuximab. The higher AKT phosphorylation level was associated with resistance to anti-EGFR agents. Therefore, we further performed drug combinations with anti-AKT agent (MK2206) and AKT1 gene editing, which demonstrated afatinib and allitinib sensitivity restored. Additionally, in silico analysis of TCGA database showed that AKT1 overexpression was present in 14.7% (41/279) of HNSCC cases, and was associated with perineural invasion in advanced stage. In conclusion, allitinib presented a greater cytotoxic profile when compared to afatinib and cetuximab. AKT pathway constitutes a predictive marker of allitinib response and combination with AKT inhibitors could restore response and increase treatment success.