Targeting focal adhesion kinase overcomes erlotinib resistance in smoke induced lung cancer by altering phosphorylation of epidermal growth factor receptor

Hitendra S. Solanki1,2,*, Remya Raja1,*, Alex Zhavoronkov3, Ivan V. Ozerov3, Artem V. Artemov3, Jayshree Advani1,4, Aneesha Radhakrishnan1, Niraj Babu1,4, Vinuth N. Puttamallesh1,5, Nazia Syed1, Vishalakshi Nanjappa1, Tejaswini Subbannayya1, Nandini A. Sahasrabuddhe1, Arun H. Patil1,2,6, T.S. Keshava Prasad1,6,7, Daria Gaykalova8, Xiaofei Chang8, Rachana Sathyendran9, Premendu Prakash Mathur2, Annapoorni Rangarajan9, David Sidransky8, Akhilesh Pandey10,11,12,13, Evgeny Izumchenko8, Harsha Gowda1,6, Aditi Chatterjee1,6

1 Institute of Bioinformatics, International Tech Park, Bangalore 560066, India

2 School of Biotechnology, Kalinga Institute of Industrial Technology, Bhubaneswar, Odisha 751024, India

3 Insilico Medicine, Inc., Emerging Technology Centers, Johns Hopkins University at Eastern, Baltimore, MD 21218, USA

4 Manipal Academy of Higher Education, Manipal, Karnataka 576104, India

5School of Biotechnology, Amrita University, Kollam 690525, India

6Center for Systems Biology and Molecular Medicine, Yenepoya (Deemed to be University), Mangalore 575018, India

7NIMHANS-IOB Proteomics and Bioinformatics Laboratory, Neurobiology Research Centre, National Institute of Mental Health and Neurosciences, Bangalore 560029, India

8Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA

9Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore, 560012, India

10McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, US

11Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA

12Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA

13Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA

*These authors contributed equally to the manuscript

Correspondence to:

Aditi Chatterjee or Harsha Gowda, email: aditi@ibioinformatics.org or harsha@ibioinformatics.org

Keywords: cigarette smoke; epidermal growth factor receptor; NSCLC; phosphoproteomics; drug resistance

Received: July 29, 2017 Accepted: December 15, 2017 Published: February 23, 2018


EGFR-based targeted therapies have shown limited success in smokers. Identification of alternate signaling mechanism(s) leading to TKI resistance in smokers is critically important. We observed increased resistance to erlotinib in H358 NSCLC (non-small cell lung carcinoma) cells chronically exposed to cigarette smoke (H358-S) compared to parental cells. SILAC-based mass-spectrometry approach was used to study altered signaling in H358-S cell line. Importantly, among the top phosphosites in H358-S cells we observed hyperphosphorylation of EGFR (Y1197) and non-receptor tyrosine kinase FAK (Y576/577). Supporting these observations, a transcriptomic-based pathway activation analysis of TCGA NSCLC datasets revealed that FAK and EGFR internalization pathways were significantly upregulated in smoking patients, compared to the never-smokers and were associated with elevated PI3K signaling and lower level of caspase cascade and E-cadherin pathways activation. We show that inhibition of FAK led to decreased cellular proliferation and invasive ability of the smoke-exposed cells, and restored their dependency on EGFR signaling. Our data suggests that activation of focal adhesion pathway significantly contributes to erlotinib resistance, and that FAK is a potential therapeutic target for management of erlotinib resistance in smoke-induced NSCLC.

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