Phosphoproteomic Analysis Identified Mutual Phosphorylation of FAK and Src as a Mechanism of Osimertinib Resistance in EGFR-Mutant Lung Cancer

Takehiro Tozuka, MD; Rintaro Noro, MD, PhD; Keisuke Yoshida, PhD; Satoshi Takahashi, MD, PhD; Mariko Hirao, MD; Kuniko Matsuda, MD; Yasuhiro Kato, MD; Shinji Nakamichi, MD, PhD; Susumu Takeuchi, MD, PhD; Masaru Matsumoto, MD, PhD; Akihiko Miyanaga, MD, PhD; Shinobu Kunugi, MD, PhD; Kazufumi Honda, DDS, PhD; Jun Adachi, PhD; Masahiro Seike, MD, PhD

JTO Clinical and Research Reports (Apr 2024)

Phosphoproteomic Analysis Identified Mutual Phosphorylation of FAK and Src as a Mechanism of Osimertinib Resistance in EGFR-Mutant Lung Cancer

Takehiro Tozuka, MD,
Rintaro Noro, MD, PhD,
Keisuke Yoshida, PhD,
Satoshi Takahashi, MD, PhD,
Mariko Hirao, MD,
Kuniko Matsuda, MD,
Yasuhiro Kato, MD,
Shinji Nakamichi, MD, PhD,
Susumu Takeuchi, MD, PhD,
Masaru Matsumoto, MD, PhD,
Akihiko Miyanaga, MD, PhD,
Shinobu Kunugi, MD, PhD,
Kazufumi Honda, DDS, PhD,
Jun Adachi, PhD,
Masahiro Seike, MD, PhD

Affiliations

Takehiro Tozuka, MD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Rintaro Noro, MD, PhD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Keisuke Yoshida, PhD: Department of Bioregulation, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Satoshi Takahashi, MD, PhD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Mariko Hirao, MD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Kuniko Matsuda, MD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Yasuhiro Kato, MD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Shinji Nakamichi, MD, PhD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Susumu Takeuchi, MD, PhD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Masaru Matsumoto, MD, PhD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Akihiko Miyanaga, MD, PhD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
Shinobu Kunugi, MD, PhD: Department of Analytic Human Pathology, Nippon Medical School, Tokyo, Japan
Kazufumi Honda, DDS, PhD: Department of Bioregulation, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan; Institution for Advanced Medical Science, Nippon Medical School, Tokyo, Japan
Jun Adachi, PhD: Laboratory of Proteomics for Drug Discovery, Center for Drug Design Research, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan
Masahiro Seike, MD, PhD: Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan; Corresponding author. Address for correspondence: Masahiro Seike, MD, PhD, Department of Pulmonary Medicine and Oncology, Graduate School of Medicine, Nippon Medical School, 1-1-5, Sendagi, Bunkyo-ku, Tokyo, 113-8603, Japan.

Journal volume & issue: Vol. 5, no. 4
p. 100668

Abstract

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Introduction: Osimertinib is a standard treatment for patients with EGFR-mutant NSCLC. Although some osimertinib resistance mechanisms have been identified, nearly 50% of the mechanisms remain to be elucidated. This study was aimed at identifying non-genetic mechanisms underlying osimertinib resistance. Methods: We established two osimertinib-resistant cell lines from EGFR mutation-positive PC-9 and HCC827 NSCLC cell lines (PC-9OR and HCC827OR, respectively) using a stepwise method. We compared the phosphoproteomic profiles of the osimertinib-resistant and parental cells using mass spectrometry. Upstream kinases were identified using the application Kinase Enrichment Analysis version 3. Results: Phosphoproteomic analysis revealed 80 phosphorylation sites that were mutually up-regulated in PC-9OR and HCC827OR cells. The Kinase Enrichment Analysis version 3 analysis identified focal adhesion kinase (FAK) and proto-oncogene tyrosine-protein kinase Src (Src) as upstream kinases of these up-regulated phosphoproteins. The small-interfering RNA–mediated knockdown of FAK reduced Src phosphorylation and that of Src reduced FAK phosphorylation in both cell lines. Furthermore, FAK- or Src-specific small-interfering RNA treatments restored EGFR phosphorylation in PC-9OR and HCC827OR cells. The combination of FAK and Src inhibitors inhibited PC-9OR and HCC827OR cell proliferation in vitro and suppressed tumor growth in a xenograft mouse model. Immunohistochemistry of tumors from patients with EGFR-mutant NSCLC suggested that phosphorylated FAK and Src are involved in initial and acquired resistance to osimertinib. Conclusions: Phosphoproteomic analysis may help elucidate the mechanisms of resistance to molecular-targeted therapies in lung cancer. Mutual phosphorylation of FAK and Src is involved in osimertinib resistance. Thus, FAK and Src inhibition may be novel treatment strategies for osimertinib-resistant NSCLC.

Published in JTO Clinical and Research Reports

ISSN: 2666-3643 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Medicine: Internal medicine: Neoplasms. Tumors. Oncology. Including cancer and carcinogens
Website: https://www.journals.elsevier.com/jto-clinical-and-research-reports/

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