Dual inhibition of GTP-bound KRAS and mTOR in lung adenocarcinoma and squamous cell carcinoma harboring KRAS G12C

Abstract Background Kirsten rat sarcoma (KRAS) mutations are somatic variants in lung adenocarcinoma. One of the most prevalent mutations, G12C, has led to the clinical approval of targeted inhibitors for advanced stages in lung cancer. Research has increasingly focused on the efficacy of combinatio...

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Main Authors: Masaoki Ito, Yoshihiro Miyata, Shoko Hirano, Nagisa Morihara, Misako Takemoto, Fumiko Irisuna, Kei Kushitani, Kenichi Suda, Junichi Soh, Yukio Takeshima, Yasuhiro Tsutani, Morihito Okada
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Language:English
Published: BMC 2025-05-01
Series:Cell Communication and Signaling
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Online Access:https://doi.org/10.1186/s12964-025-02187-y
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author Masaoki Ito
Yoshihiro Miyata
Shoko Hirano
Nagisa Morihara
Misako Takemoto
Fumiko Irisuna
Kei Kushitani
Kenichi Suda
Junichi Soh
Yukio Takeshima
Yasuhiro Tsutani
Morihito Okada
author_facet Masaoki Ito
Yoshihiro Miyata
Shoko Hirano
Nagisa Morihara
Misako Takemoto
Fumiko Irisuna
Kei Kushitani
Kenichi Suda
Junichi Soh
Yukio Takeshima
Yasuhiro Tsutani
Morihito Okada
author_sort Masaoki Ito
collection DOAJ
description Abstract Background Kirsten rat sarcoma (KRAS) mutations are somatic variants in lung adenocarcinoma. One of the most prevalent mutations, G12C, has led to the clinical approval of targeted inhibitors for advanced stages in lung cancer. Research has increasingly focused on the efficacy of combination therapies that target multiple tumorigenic pathways. Cases harboring KRAS G12C mutation are heterogenous. We explored alternative changes in genetic pathways and evaluated the effectiveness of combination therapy using several types of cell lines and KRAS inhibitors. Methods We comprehensively investigated genetic changes induced by KRAS G12C inhibition using RNA sequences and the candidate to inhibit in combination therapy was explored. Three lung cancer cell lines (two adenocarcinoma and one squamous cell carcinoma) and three KRAS G12C inhibitors (AMG 510, MRTX849, and ARS-1620) were used. KRAS G12C and candidate gene were simultaneously inhibited in cell lines and the efficiency of combination therapy was evaluated using clonogenic assays and MTS assay. Pathway activation was assessed via western blotting. A combination index (CI) < 0.8 was considered statistically synergistic. Results RNA sequences revealed treatment with two of the three KRAS G12C inhibitors led to a significant increase in mTOR expression across all three cell lines. mTOR was targeted in combination therapy; each KRAS G12C inhibitor and mTOR inhibitor (RAD001) combination exhibited synergism (CI < 0.8) in MTS and clonogenic assays. Single inhibition of mTOR induced activation of guanosine triphosphate (GTP)-RAS, thereby activating the RAS-MEK-ERK and PI3K-AKT-mTOR pathways in WB, suggesting mTOR activation is crucial for KRAS-driving lung cancer. A combination strategy targeting KRAS G12C and mTOR abrogated GTP-RAS, pmTOR (Ser2448), and pERK (Thr202/Tyr204) more efficiently. Conclusions KRAS G12C inhibitor plus RAD001 consistently revealed synergism. Targeting KRAS G12C and mTOR abrogates the RAS-MEK-ERK and PI3K-AKT-mTOR pathways. Our data suggests that a combined strategy targeting GTP-bound KRAS G12C and mTOR shows promise for primary lung cancers with KRAS G12C mutations. This approach may also be effective even for lung cancers harboring KRAS G12C mutation but having different profiles.
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spelling doaj-art-53b6f7b1b143402eb378e00cc15e5d8a2025-08-20T03:10:31ZengBMCCell Communication and Signaling1478-811X2025-05-0123111110.1186/s12964-025-02187-yDual inhibition of GTP-bound KRAS and mTOR in lung adenocarcinoma and squamous cell carcinoma harboring KRAS G12CMasaoki Ito0Yoshihiro Miyata1Shoko Hirano2Nagisa Morihara3Misako Takemoto4Fumiko Irisuna5Kei Kushitani6Kenichi Suda7Junichi Soh8Yukio Takeshima9Yasuhiro Tsutani10Morihito Okada11Department of Surgical Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima UniversityDepartment of Surgical Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima UniversityAnalysis Center of Life Science, Natural Science Center for Basic Research and Development, Hiroshima UniversityAnalysis Center of Life Science, Natural Science Center for Basic Research and Development, Hiroshima UniversityAnalysis Center of Life Science, Natural Science Center for Basic Research and Development, Hiroshima UniversityAnalysis Center of Life Science, Natural Science Center for Basic Research and Development, Hiroshima UniversityDepartment of Pathology, Graduate School of Biomedical & Health Sciences, Hiroshima UniversityDivision of Thoracic Surgery, Department of Surgery, Kindai University Faculty of MedicineDivision of Thoracic Surgery, Department of Surgery, Kindai University Faculty of MedicineDepartment of Pathology, Graduate School of Biomedical & Health Sciences, Hiroshima UniversityDivision of Thoracic Surgery, Department of Surgery, Kindai University Faculty of MedicineDepartment of Surgical Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima UniversityAbstract Background Kirsten rat sarcoma (KRAS) mutations are somatic variants in lung adenocarcinoma. One of the most prevalent mutations, G12C, has led to the clinical approval of targeted inhibitors for advanced stages in lung cancer. Research has increasingly focused on the efficacy of combination therapies that target multiple tumorigenic pathways. Cases harboring KRAS G12C mutation are heterogenous. We explored alternative changes in genetic pathways and evaluated the effectiveness of combination therapy using several types of cell lines and KRAS inhibitors. Methods We comprehensively investigated genetic changes induced by KRAS G12C inhibition using RNA sequences and the candidate to inhibit in combination therapy was explored. Three lung cancer cell lines (two adenocarcinoma and one squamous cell carcinoma) and three KRAS G12C inhibitors (AMG 510, MRTX849, and ARS-1620) were used. KRAS G12C and candidate gene were simultaneously inhibited in cell lines and the efficiency of combination therapy was evaluated using clonogenic assays and MTS assay. Pathway activation was assessed via western blotting. A combination index (CI) < 0.8 was considered statistically synergistic. Results RNA sequences revealed treatment with two of the three KRAS G12C inhibitors led to a significant increase in mTOR expression across all three cell lines. mTOR was targeted in combination therapy; each KRAS G12C inhibitor and mTOR inhibitor (RAD001) combination exhibited synergism (CI < 0.8) in MTS and clonogenic assays. Single inhibition of mTOR induced activation of guanosine triphosphate (GTP)-RAS, thereby activating the RAS-MEK-ERK and PI3K-AKT-mTOR pathways in WB, suggesting mTOR activation is crucial for KRAS-driving lung cancer. A combination strategy targeting KRAS G12C and mTOR abrogated GTP-RAS, pmTOR (Ser2448), and pERK (Thr202/Tyr204) more efficiently. Conclusions KRAS G12C inhibitor plus RAD001 consistently revealed synergism. Targeting KRAS G12C and mTOR abrogates the RAS-MEK-ERK and PI3K-AKT-mTOR pathways. Our data suggests that a combined strategy targeting GTP-bound KRAS G12C and mTOR shows promise for primary lung cancers with KRAS G12C mutations. This approach may also be effective even for lung cancers harboring KRAS G12C mutation but having different profiles.https://doi.org/10.1186/s12964-025-02187-yKRAS G12CmTORCombination therapyLung adenocarcinomaSquamous cell carcinomaSotorasib
spellingShingle Masaoki Ito
Yoshihiro Miyata
Shoko Hirano
Nagisa Morihara
Misako Takemoto
Fumiko Irisuna
Kei Kushitani
Kenichi Suda
Junichi Soh
Yukio Takeshima
Yasuhiro Tsutani
Morihito Okada
Dual inhibition of GTP-bound KRAS and mTOR in lung adenocarcinoma and squamous cell carcinoma harboring KRAS G12C
Cell Communication and Signaling
KRAS G12C
mTOR
Combination therapy
Lung adenocarcinoma
Squamous cell carcinoma
Sotorasib
title Dual inhibition of GTP-bound KRAS and mTOR in lung adenocarcinoma and squamous cell carcinoma harboring KRAS G12C
title_full Dual inhibition of GTP-bound KRAS and mTOR in lung adenocarcinoma and squamous cell carcinoma harboring KRAS G12C
title_fullStr Dual inhibition of GTP-bound KRAS and mTOR in lung adenocarcinoma and squamous cell carcinoma harboring KRAS G12C
title_full_unstemmed Dual inhibition of GTP-bound KRAS and mTOR in lung adenocarcinoma and squamous cell carcinoma harboring KRAS G12C
title_short Dual inhibition of GTP-bound KRAS and mTOR in lung adenocarcinoma and squamous cell carcinoma harboring KRAS G12C
title_sort dual inhibition of gtp bound kras and mtor in lung adenocarcinoma and squamous cell carcinoma harboring kras g12c
topic KRAS G12C
mTOR
Combination therapy
Lung adenocarcinoma
Squamous cell carcinoma
Sotorasib
url https://doi.org/10.1186/s12964-025-02187-y
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