Abstract 3995: Exploring non-small cell lung carcinoma cell lines' sensitivity to neratinib
Authors: Owen Sioned , Dart Dafydd A. , Lalani Alshad S. , Avogadri-Connors Francesca , Bryce Richard P. , Jiang Wen G. 
Published: August 2018

Introduction: Lung cancer is the second most common diagnosed malignancy and accounts for the biggest number of cancer related deaths worldwide. Despite increased health warnings associated with habits, such as smoking, ten year survival rates for lung cancer in the UK remain similar to those seen in the 1970's. In the USA, the 5 year survival rates dramatically reduce with late disease stage, from 49 percent at Stage 1A to 5 percent at Stage IIIB. Mutations in epidermal growth factor receptor (EGFR) appears to be quite common in lung cancer. ERBB2 mutations also appear, albeit at lower frequency. In addition, ERBB2 amplifications have also been reported, particularly in tumors that have become resistant to EGFR-TKIs. Neratinib is an orally available tyrosine kinase inhibitor that irreversibly binds and inhibits EGFR, ERBB2 and ERBB4 receptor tyrosine kinases. This study aimed to characterize in detail the expression profile of genes downstream of the EGFR family members in two NSCLC cell lines and their differential sensitivity to neratinib. Materials and Methods: A549 (adenocarcinoma), a cell line with intermediate sensitivity to gefitnib, and SKMES-1 (squamous cell carcinoma) NSCLC cell lines were purchased from American Tissue Culture Collection (ATCC). Cytotoxicity was assessed using an MTT assay. Changes in gene expression in response to neratinib were quantified by next generation sequencing using AmpliSeq technology and analysed using Ingenuity pathway analysis (IPA) software.

Results: SKMES-1 lung cancer cells were found to be more sensitive to neratinib than A549 cancer cells. Both cell lines exhibited similar low gene expression levels of ERBB2. SKMES-1 cancer cells showed higher gene expression profiles for the remaining HER family members, EGFR, ERBB3 and ERBB4, compared to the A549 cancer cells. IPA analysis showed that in SKMES-1 cells treated with neratinib differentially expressed genes were associated with inhibition of Akt and ERK pathways and their signal transduction whereas, this was not seen in the A549 cells.

Conclusions: This study has shown that although these two NSCLC cell lines have similar gene expression levels of ERBB2, there is a difference in profile amongst the other receptor family members. EGFR and ERBB4 mRNA transcript levels were twice as high in SK-MES-1 compared to A549. This variance in the ERBB family gene expression profile may account for the differing sensitivities these cell lines have to neratinib. Further analysis indicates that Akt/ERK signal transduction was inhibited in SKMES-1 cells treated with neratinib though the exact mechanism for this remains unknown.

Citation Format: Sioned Owen, Dafydd A. Dart, Alshad S. Lalani, Francesca Avogadri-Connors, Richard P. Bryce, Wen G. Jiang. Exploring non-small cell lung carcinoma cell lines' sensitivity to neratinib [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3995.