TKI-addicted ROS1-rearranged cells are destined to survival or death by the intensity of ROS1 kinase activity

• Published in: Scientific reports Vol. 7; no. 1; pp. 5519 - 17
• Main Authors: Ogura, Hayato, Nagatake-Kobayashi, Yuka, Adachi, Jun, Tomonaga, Takeshi, Fujita, Naoya, Katayama, Ryohei
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 17-07-2017; Nature Publishing Group; Nature Portfolio
• Subjects: 13; 13/109; 692/4028/67/1059/602; 692/4028/67/1612/1350; 82/58; 82/80; 96/31; 96/98; Anilides - pharmacology; Antigens, Differentiation, B-Lymphocyte - genetics; Antigens, Differentiation, B-Lymphocyte - metabolism; Apoptosis; Apoptosis - drug effects; Cell Line, Tumor; Doxycycline; Doxycycline - pharmacology; Drug Resistance, Neoplasm - genetics; Enzyme inhibitors; Ethyl nitrosourea; Ethylnitrosourea - pharmacology; Histocompatibility Antigens Class II - genetics; Histocompatibility Antigens Class II - metabolism; Humanities and Social Sciences; Humans; Imidazoles - pharmacology; Kinases; Lung cancer; Molecular modelling; multidisciplinary; Mutagenesis; Mutagenesis, Site-Directed; Mutation; Non-small cell lung carcinoma; Overexpression; p38 Mitogen-Activated Protein Kinases - antagonists & inhibitors; p38 Mitogen-Activated Protein Kinases - metabolism; Phosphopeptides - analysis; Protein Kinase Inhibitors - pharmacology; Protein Structure, Tertiary; Protein-tyrosine kinase; Protein-Tyrosine Kinases - chemistry; Protein-Tyrosine Kinases - genetics; Protein-Tyrosine Kinases - metabolism; Proto-Oncogene Proteins - chemistry; Proto-Oncogene Proteins - genetics; Proto-Oncogene Proteins - metabolism; Pyridines - pharmacology; Science; Science (multidisciplinary); Signal Transduction - drug effects; Tandem Mass Spectrometry; Tumors
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-017-05736-9

ROS1
rearrangement is observed in 1–2% of non-small cell lung cancers (NSCLC). The ROS1 tyrosine kinase inhibitor (TKI) crizotinib has induced marked tumour shrinkage in
ROS1
-rearranged cancers. However, emergence of acquired resistance to TKI is inevitable within a few years. Previous findings indicate that cabozantinib overcomes secondary mutation–mediated crizotinib-resistance in
ROS1
-fusion-positive cells. Here we attempted to establish cabozantinib-resistant cells by
N
-ethyl-
N
-nitrosourea mutagenesis screening using CD74-ROS1–expressing Ba/F3 cells. Two resistant cell lines with CD74-ROS1 F2004V or F2075C mutations, which are homologous to ALK F1174 or F1245 mutations, survived in the presence of a low dose of ROS1-TKI. Removal of ROS1-TKI from these TKI-addicted cells induced excessive activation of ROS1 tyrosine kinase followed by apoptosis. We succeeded in recapturing the TKI-addicted phenotype using doxycycline-inducible CD74-ROS1 mutant over-expression in Ba/F3 cells, suggesting that excessive
ROS1
oncogenic signaling itself induced apoptosis instead of cell growth. Phosphoproteomic analysis and high-throughput inhibitor screening revealed that excessive ROS1 signaling in the TKI-addicted cells phosphorylated or activated apoptosis-related molecules such as FAF1 or p38. Collectively, our findings partly clarify molecular mechanisms of excessive
ROS1
oncogenic signaling that mediates paradoxical induction of apoptosis.