Nutlin-3a induces KRAS mutant/p53 wild type lung cancer specific methuosis-like cell death that is dependent on GFPT2
Abstract Background Oncogenic KRAS mutation, the most frequent mutation in non-small cell lung cancer (NSCLC), is an aggressiveness risk factor and leads to the metabolic reprogramming of cancer cells by promoting glucose, glutamine, and fatty acid absorption and glycolysis. Lately, sotorasib was ap...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-12-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13046-023-02922-8 |
| _version_ | 1797388008973926400 |
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| author | Dasom Kim Dongwha Min Joohee Kim Min Jung Kim Yerim Seo Byung Hwa Jung Seung‑Hae Kwon Hyunju Ro Seoee Lee Jason K. Sa Ji-Yun Lee |
| author_facet | Dasom Kim Dongwha Min Joohee Kim Min Jung Kim Yerim Seo Byung Hwa Jung Seung‑Hae Kwon Hyunju Ro Seoee Lee Jason K. Sa Ji-Yun Lee |
| author_sort | Dasom Kim |
| collection | DOAJ |
| description | Abstract Background Oncogenic KRAS mutation, the most frequent mutation in non-small cell lung cancer (NSCLC), is an aggressiveness risk factor and leads to the metabolic reprogramming of cancer cells by promoting glucose, glutamine, and fatty acid absorption and glycolysis. Lately, sotorasib was approved by the FDA as a first-in-class KRAS-G12C inhibitor. However, sotorasib still has a derivative barrier, which is not effective for other KRAS mutation types, except for G12C. Additionally, resistance to sotorasib is likely to develop, demanding the need for alternative therapeutic strategies. Methods KRAS mutant, and wildtype NSCLC cells were used in vitro cell analyses. Cell viability, proliferation, and death were measured by MTT, cell counting, colony analyses, and annexin V staining for FACS. Cell tracker dyes were used to investigate cell morphology, which was examined by holotomograpy, and confocal microscopes. RNA sequencing was performed to identify key target molecule or pathway, which was confirmed by qRT-PCR, western blotting, and metabolite analyses by UHPLC-MS/MS. Zebrafish and mouse xenograft model were used for in vivo analysis. Results In this study, we found that nutlin-3a, an MDM2 antagonist, inhibited the KRAS-PI3K/Akt-mTOR pathway and disrupted the fusion of both autophagosomes and macropinosomes with lysosomes. This further elucidated non-apoptotic and catastrophic macropinocytosis associated methuosis-like cell death, which was found to be dependent on GFPT2 of the hexosamine biosynthetic pathway, specifically in KRAS mutant /p53 wild type NSCLC cells. Conclusion These results indicate the potential of nutlin-3a as an alternative agent for treating KRAS mutant/p53 wild type NSCLC cells. |
| first_indexed | 2024-03-08T22:34:29Z |
| format | Article |
| id | doaj.art-9eae5048fa874e669d3b8525faa03c5e |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-03-08T22:34:29Z |
| publishDate | 2023-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-9eae5048fa874e669d3b8525faa03c5e2023-12-17T12:33:40ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662023-12-0142111710.1186/s13046-023-02922-8Nutlin-3a induces KRAS mutant/p53 wild type lung cancer specific methuosis-like cell death that is dependent on GFPT2Dasom Kim0Dongwha Min1Joohee Kim2Min Jung Kim3Yerim Seo4Byung Hwa Jung5Seung‑Hae Kwon6Hyunju Ro7Seoee Lee8Jason K. Sa9Ji-Yun Lee10Department of Pathology, Korea University College of MedicineDepartment of Pathology, Korea University College of MedicineDepartment of Biological Sciences, Sookmyung Women’s UniversityDepartment of Biological Sciences, Sookmyung Women’s UniversityCenter for Advanced Biomolecular Recognition, Korea Instiute of Science and Technology (KIST)Center for Advanced Biomolecular Recognition, Korea Instiute of Science and Technology (KIST)Korea Basic Science Institute, Seoul CenterDepartment of Biological Sciences, College of Bioscience and Biotechnology, Chungnam National UniversityDepartment of Biological Sciences, College of Bioscience and Biotechnology, Chungnam National UniversityDepartment of Biomedical Sciences, Korea University College of MedicineDepartment of Pathology, Korea University College of MedicineAbstract Background Oncogenic KRAS mutation, the most frequent mutation in non-small cell lung cancer (NSCLC), is an aggressiveness risk factor and leads to the metabolic reprogramming of cancer cells by promoting glucose, glutamine, and fatty acid absorption and glycolysis. Lately, sotorasib was approved by the FDA as a first-in-class KRAS-G12C inhibitor. However, sotorasib still has a derivative barrier, which is not effective for other KRAS mutation types, except for G12C. Additionally, resistance to sotorasib is likely to develop, demanding the need for alternative therapeutic strategies. Methods KRAS mutant, and wildtype NSCLC cells were used in vitro cell analyses. Cell viability, proliferation, and death were measured by MTT, cell counting, colony analyses, and annexin V staining for FACS. Cell tracker dyes were used to investigate cell morphology, which was examined by holotomograpy, and confocal microscopes. RNA sequencing was performed to identify key target molecule or pathway, which was confirmed by qRT-PCR, western blotting, and metabolite analyses by UHPLC-MS/MS. Zebrafish and mouse xenograft model were used for in vivo analysis. Results In this study, we found that nutlin-3a, an MDM2 antagonist, inhibited the KRAS-PI3K/Akt-mTOR pathway and disrupted the fusion of both autophagosomes and macropinosomes with lysosomes. This further elucidated non-apoptotic and catastrophic macropinocytosis associated methuosis-like cell death, which was found to be dependent on GFPT2 of the hexosamine biosynthetic pathway, specifically in KRAS mutant /p53 wild type NSCLC cells. Conclusion These results indicate the potential of nutlin-3a as an alternative agent for treating KRAS mutant/p53 wild type NSCLC cells.https://doi.org/10.1186/s13046-023-02922-8KRAS mutationGFPT2HBPMacropinocytosisNutlin-3aMethuosis |
| spellingShingle | Dasom Kim Dongwha Min Joohee Kim Min Jung Kim Yerim Seo Byung Hwa Jung Seung‑Hae Kwon Hyunju Ro Seoee Lee Jason K. Sa Ji-Yun Lee Nutlin-3a induces KRAS mutant/p53 wild type lung cancer specific methuosis-like cell death that is dependent on GFPT2 Journal of Experimental & Clinical Cancer Research KRAS mutation GFPT2 HBP Macropinocytosis Nutlin-3a Methuosis |
| title | Nutlin-3a induces KRAS mutant/p53 wild type lung cancer specific methuosis-like cell death that is dependent on GFPT2 |
| title_full | Nutlin-3a induces KRAS mutant/p53 wild type lung cancer specific methuosis-like cell death that is dependent on GFPT2 |
| title_fullStr | Nutlin-3a induces KRAS mutant/p53 wild type lung cancer specific methuosis-like cell death that is dependent on GFPT2 |
| title_full_unstemmed | Nutlin-3a induces KRAS mutant/p53 wild type lung cancer specific methuosis-like cell death that is dependent on GFPT2 |
| title_short | Nutlin-3a induces KRAS mutant/p53 wild type lung cancer specific methuosis-like cell death that is dependent on GFPT2 |
| title_sort | nutlin 3a induces kras mutant p53 wild type lung cancer specific methuosis like cell death that is dependent on gfpt2 |
| topic | KRAS mutation GFPT2 HBP Macropinocytosis Nutlin-3a Methuosis |
| url | https://doi.org/10.1186/s13046-023-02922-8 |
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