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

Abstract 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 find...

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Main Authors: Hayato Ogura, Yuka Nagatake-Kobayashi, Jun Adachi, Takeshi Tomonaga, Naoya Fujita, Ryohei Katayama
Format: Article
Language:English
Published: Nature Publishing Group 2017-07-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-017-05736-9
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spelling doaj-c2b81b6512f34d54906db2bec495e3632020-12-08T00:08:17ZengNature Publishing GroupScientific Reports2045-23222017-07-017111710.1038/s41598-017-05736-9TKI-addicted ROS1-rearranged cells are destined to survival or death by the intensity of ROS1 kinase activityHayato Ogura0Yuka Nagatake-Kobayashi1Jun Adachi2Takeshi Tomonaga3Naoya Fujita4Ryohei Katayama5Division of Experimental Chemotherapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer ResearchDivision of Experimental Chemotherapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer ResearchLaboratory of Proteome Research, National Institute of Biomedical Innovation, Health and NutritionLaboratory of Proteome Research, National Institute of Biomedical Innovation, Health and NutritionDivision of Experimental Chemotherapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer ResearchDivision of Experimental Chemotherapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer ResearchAbstract 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.https://doi.org/10.1038/s41598-017-05736-9
collection DOAJ
language English
format Article
sources DOAJ
author Hayato Ogura
Yuka Nagatake-Kobayashi
Jun Adachi
Takeshi Tomonaga
Naoya Fujita
Ryohei Katayama
spellingShingle Hayato Ogura
Yuka Nagatake-Kobayashi
Jun Adachi
Takeshi Tomonaga
Naoya Fujita
Ryohei Katayama
TKI-addicted ROS1-rearranged cells are destined to survival or death by the intensity of ROS1 kinase activity
Scientific Reports
author_facet Hayato Ogura
Yuka Nagatake-Kobayashi
Jun Adachi
Takeshi Tomonaga
Naoya Fujita
Ryohei Katayama
author_sort Hayato Ogura
title TKI-addicted ROS1-rearranged cells are destined to survival or death by the intensity of ROS1 kinase activity
title_short TKI-addicted ROS1-rearranged cells are destined to survival or death by the intensity of ROS1 kinase activity
title_full TKI-addicted ROS1-rearranged cells are destined to survival or death by the intensity of ROS1 kinase activity
title_fullStr TKI-addicted ROS1-rearranged cells are destined to survival or death by the intensity of ROS1 kinase activity
title_full_unstemmed TKI-addicted ROS1-rearranged cells are destined to survival or death by the intensity of ROS1 kinase activity
title_sort tki-addicted ros1-rearranged cells are destined to survival or death by the intensity of ros1 kinase activity
publisher Nature Publishing Group
series Scientific Reports
issn 2045-2322
publishDate 2017-07-01
description Abstract 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.
url https://doi.org/10.1038/s41598-017-05736-9
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