CT45A1‐mediated MLC2 (MYL9) phosphorylation promotes natural killer cell resistance and outer cell fate in a cell‐in‐cell structure, potentiating the progression of microsatellite instability‐high colorectal cancer
Patients with microsatellite instability‐high (MSI‐H) colorectal cancer (CRC) have high tumor mutation burden and tumor immunogenicity, exhibiting a higher response rate to immunotherapy and better survival. However, a portion of MSI‐H CRC patients still experience adverse disease outcomes. We aimed...
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| Format: | Article |
| Language: | English |
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Wiley
2025-02-01
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| Series: | Molecular Oncology |
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| Online Access: | https://doi.org/10.1002/1878-0261.13736 |
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| author | Hao‐Wei Teng Hsiang‐Yueh Huang Chun‐Chi Lin Yuh‐Ching Twu Wen‐Hao Yang Wen‐Chun Lin Hsin‐Yi Lan Yen‐Yu Lin Wei‐Lun Hwang |
| author_facet | Hao‐Wei Teng Hsiang‐Yueh Huang Chun‐Chi Lin Yuh‐Ching Twu Wen‐Hao Yang Wen‐Chun Lin Hsin‐Yi Lan Yen‐Yu Lin Wei‐Lun Hwang |
| author_sort | Hao‐Wei Teng |
| collection | DOAJ |
| description | Patients with microsatellite instability‐high (MSI‐H) colorectal cancer (CRC) have high tumor mutation burden and tumor immunogenicity, exhibiting a higher response rate to immunotherapy and better survival. However, a portion of MSI‐H CRC patients still experience adverse disease outcomes. We aimed to identify the tumor‐autonomous regulators determining these heterogeneous clinical outcomes. The Cancer Genome Atlas (TCGA) dataset was used to identify regulators in MSI‐H CRC patients with unfavorable outcomes. Stable CRC tumor clones expressing targeted regulators were established to evaluate migratory and stemness properties, immune cell vulnerability, and cell‐in‐cell (CIC) structure formation. RNA‐sequencing (RNA‐seq) was used to identify enriched biological pathways in stable CRC tumor clones. Clinicopathological characterization of formalin‐fixed paraffin‐embedded (FFPE) MSI‐H CRC specimens was performed to explore the underlying mechanisms involved. We showed that cancer/testis antigen family 45 member A1 (CT45A1) expression was upregulated in MSI‐H CRC patients with poor survival outcomes. CT45A1‐expressing microsatellite stable (MSS) CRC cells showed enhanced migratory ability. However, CT45A1‐expressing MSI‐H CRC cells, but not MSS CRC cells, showed higher resistance to natural killer (NK) cell cytotoxicity and served as outer cells in homotypic CIC structures, preventing exogenous or therapeutic antibody access to inner CRC cells. Inactivating RHO‐ROCK/MLCK‐MLC2 signaling with small‐molecule inhibitors or short‐hairpin RNAs (shRNAs) targeting myosin light chain kinase (MYLK) abolished NK cell resistance and reduced the outer cell fate of CT45A1‐expressing MSI‐H CRC cells. In MSI‐H CRC patients, CT45A1‐positive tumors exhibited increased MLC2 phosphorylation, increased outer cell fate, and decreased survival. We demonstrated that CT45A1 potentiates the advanced progression of MSI‐H CRC, and targeting MLC2 phosphorylation may enhance immunotherapy efficacy in CT45A1‐positive MSI‐H CRC patients. |
| format | Article |
| id | doaj-art-947287fb253e4d1bac2c6c53a7b2bc4d |
| institution | Kabale University |
| issn | 1574-7891 1878-0261 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Molecular Oncology |
| spelling | doaj-art-947287fb253e4d1bac2c6c53a7b2bc4d2025-02-04T17:30:20ZengWileyMolecular Oncology1574-78911878-02612025-02-0119243045110.1002/1878-0261.13736CT45A1‐mediated MLC2 (MYL9) phosphorylation promotes natural killer cell resistance and outer cell fate in a cell‐in‐cell structure, potentiating the progression of microsatellite instability‐high colorectal cancerHao‐Wei Teng0Hsiang‐Yueh Huang1Chun‐Chi Lin2Yuh‐Ching Twu3Wen‐Hao Yang4Wen‐Chun Lin5Hsin‐Yi Lan6Yen‐Yu Lin7Wei‐Lun Hwang8Division of Medical Oncology, Department of Oncology Taipei Veterans General Hospital TaiwanDepartment of Biotechnology and Laboratory Science in Medicine National Yang Ming Chiao Tung University Taipei TaiwanDivision of Colon and Rectum Surgery, Department of Surgery Taipei Veterans General Hospital TaiwanDepartment of Biotechnology and Laboratory Science in Medicine National Yang Ming Chiao Tung University Taipei TaiwanGraduate Institute of Biomedical Sciences, Research Center for Cancer Biology and Center for Molecular Medicine China Medical University Taichung TaiwanDepartment of Biotechnology and Laboratory Science in Medicine National Yang Ming Chiao Tung University Taipei TaiwanDepartment of Biotechnology and Laboratory Science in Medicine National Yang Ming Chiao Tung University Taipei TaiwanDepartment of Pathology, Fu Jen Catholic University Hospital Fu Jen Catholic University New Taipei City TaiwanDepartment of Biotechnology and Laboratory Science in Medicine National Yang Ming Chiao Tung University Taipei TaiwanPatients with microsatellite instability‐high (MSI‐H) colorectal cancer (CRC) have high tumor mutation burden and tumor immunogenicity, exhibiting a higher response rate to immunotherapy and better survival. However, a portion of MSI‐H CRC patients still experience adverse disease outcomes. We aimed to identify the tumor‐autonomous regulators determining these heterogeneous clinical outcomes. The Cancer Genome Atlas (TCGA) dataset was used to identify regulators in MSI‐H CRC patients with unfavorable outcomes. Stable CRC tumor clones expressing targeted regulators were established to evaluate migratory and stemness properties, immune cell vulnerability, and cell‐in‐cell (CIC) structure formation. RNA‐sequencing (RNA‐seq) was used to identify enriched biological pathways in stable CRC tumor clones. Clinicopathological characterization of formalin‐fixed paraffin‐embedded (FFPE) MSI‐H CRC specimens was performed to explore the underlying mechanisms involved. We showed that cancer/testis antigen family 45 member A1 (CT45A1) expression was upregulated in MSI‐H CRC patients with poor survival outcomes. CT45A1‐expressing microsatellite stable (MSS) CRC cells showed enhanced migratory ability. However, CT45A1‐expressing MSI‐H CRC cells, but not MSS CRC cells, showed higher resistance to natural killer (NK) cell cytotoxicity and served as outer cells in homotypic CIC structures, preventing exogenous or therapeutic antibody access to inner CRC cells. Inactivating RHO‐ROCK/MLCK‐MLC2 signaling with small‐molecule inhibitors or short‐hairpin RNAs (shRNAs) targeting myosin light chain kinase (MYLK) abolished NK cell resistance and reduced the outer cell fate of CT45A1‐expressing MSI‐H CRC cells. In MSI‐H CRC patients, CT45A1‐positive tumors exhibited increased MLC2 phosphorylation, increased outer cell fate, and decreased survival. We demonstrated that CT45A1 potentiates the advanced progression of MSI‐H CRC, and targeting MLC2 phosphorylation may enhance immunotherapy efficacy in CT45A1‐positive MSI‐H CRC patients.https://doi.org/10.1002/1878-0261.13736cell‐in‐cell structurecolorectal cancerCT45A1microsatellite instability‐highnatural killer cells |
| spellingShingle | Hao‐Wei Teng Hsiang‐Yueh Huang Chun‐Chi Lin Yuh‐Ching Twu Wen‐Hao Yang Wen‐Chun Lin Hsin‐Yi Lan Yen‐Yu Lin Wei‐Lun Hwang CT45A1‐mediated MLC2 (MYL9) phosphorylation promotes natural killer cell resistance and outer cell fate in a cell‐in‐cell structure, potentiating the progression of microsatellite instability‐high colorectal cancer Molecular Oncology cell‐in‐cell structure colorectal cancer CT45A1 microsatellite instability‐high natural killer cells |
| title | CT45A1‐mediated MLC2 (MYL9) phosphorylation promotes natural killer cell resistance and outer cell fate in a cell‐in‐cell structure, potentiating the progression of microsatellite instability‐high colorectal cancer |
| title_full | CT45A1‐mediated MLC2 (MYL9) phosphorylation promotes natural killer cell resistance and outer cell fate in a cell‐in‐cell structure, potentiating the progression of microsatellite instability‐high colorectal cancer |
| title_fullStr | CT45A1‐mediated MLC2 (MYL9) phosphorylation promotes natural killer cell resistance and outer cell fate in a cell‐in‐cell structure, potentiating the progression of microsatellite instability‐high colorectal cancer |
| title_full_unstemmed | CT45A1‐mediated MLC2 (MYL9) phosphorylation promotes natural killer cell resistance and outer cell fate in a cell‐in‐cell structure, potentiating the progression of microsatellite instability‐high colorectal cancer |
| title_short | CT45A1‐mediated MLC2 (MYL9) phosphorylation promotes natural killer cell resistance and outer cell fate in a cell‐in‐cell structure, potentiating the progression of microsatellite instability‐high colorectal cancer |
| title_sort | ct45a1 mediated mlc2 myl9 phosphorylation promotes natural killer cell resistance and outer cell fate in a cell in cell structure potentiating the progression of microsatellite instability high colorectal cancer |
| topic | cell‐in‐cell structure colorectal cancer CT45A1 microsatellite instability‐high natural killer cells |
| url | https://doi.org/10.1002/1878-0261.13736 |
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