Osteoclast-expanded supercharged NK cells perform superior antitumour effector functions
Objective Natural killer (NK) cells are the largest innate lymphocyte subset with potent antitumour and antiviral functions. However, clinical utilisation of human NK cells is hampered due to a lack of reliable methods to augment their antitumour potential. We demonstrated technology in which human...
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| Format: | Article |
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BMJ Publishing Group
2025-06-01
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| Series: | BMJ Oncology |
| Online Access: | https://bmjoncology.bmj.com/content/4/1/e000676.full |
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| author | Po-Chun Chen Subramaniam Malarkannan Ao Mei Kawaljit Kaur Anahid Jewett Meng-Wei Ko Emanuela Senjor Milica Perišić Nanut Lucy Wanrong Gao Paul Wong Whitaker Cohn Julian P Whitelegge Janko Kos |
| author_facet | Po-Chun Chen Subramaniam Malarkannan Ao Mei Kawaljit Kaur Anahid Jewett Meng-Wei Ko Emanuela Senjor Milica Perišić Nanut Lucy Wanrong Gao Paul Wong Whitaker Cohn Julian P Whitelegge Janko Kos |
| author_sort | Po-Chun Chen |
| collection | DOAJ |
| description | Objective Natural killer (NK) cells are the largest innate lymphocyte subset with potent antitumour and antiviral functions. However, clinical utilisation of human NK cells is hampered due to a lack of reliable methods to augment their antitumour potential. We demonstrated technology in which human NK cells were cocultured with osteoclasts in the presence of probiotic bacteria. This approach significantly augmented the antitumour cytotoxicity and polyfunctionality of human NK cells, resulting in the generation of supercharged NK (sNK) cells.Methods and analysis We explored the proteomic, transcriptomic and functional characterisation of sNK cells using cell imaging, flow cytometric analysis, 51-chromium release cytotoxicity assay, ELISA, ELIspot, IsoPLexis single-cell secretome analysis, proteomic analysis, RNA analysis, western blot and enzyme kinetics.Results We found that sNK cells were less susceptible to split anergy and tumour-induced exhaustion. Proteomic analyses revealed that sNK cells significantly increased their cell motility and proliferation. Single-cell transcriptomes uncovered sNK cells undertaking a unique differentiation trajectory and turning on STAT1, JUN, BHLHE40, ELF1, MAX and MYC regulons essential for augmenting antitumour effector functions and proliferation, respectively. Both proteomic and single-cell transcriptomes revealed that an increase in Cathepsin C helped to augment the quantity and function of Granzyme B.Conclusions These results support that this unique method produces potent NK cells for clinical utilisation and delineate the molecular mechanisms associated with this process. |
| format | Article |
| id | doaj-art-3d3847cb79ec4e5b8fdd6ae5b10e82f4 |
| institution | DOAJ |
| issn | 2752-7948 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | BMJ Publishing Group |
| record_format | Article |
| series | BMJ Oncology |
| spelling | doaj-art-3d3847cb79ec4e5b8fdd6ae5b10e82f42025-08-20T02:44:23ZengBMJ Publishing GroupBMJ Oncology2752-79482025-06-014110.1136/bmjonc-2024-000676Osteoclast-expanded supercharged NK cells perform superior antitumour effector functionsPo-Chun Chen0Subramaniam Malarkannan1Ao Mei2Kawaljit Kaur3Anahid Jewett4Meng-Wei Ko5Emanuela Senjor6Milica Perišić Nanut7Lucy Wanrong Gao8Paul Wong9Whitaker Cohn10Julian P Whitelegge11Janko Kos12Division of Oral Biology and Medicine, The Jane and Jerry Weintraub Center for Reconstructive Biotechnology, University of California Los Angeles School of Dentistry, Los Angeles, California, USADepartment of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, Wisconsin, USADepartment of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, Wisconsin, USADivision of Oral Biology and Medicine, The Jane and Jerry Weintraub Center for Reconstructive Biotechnology, University of California Los Angeles School of Dentistry, Los Angeles, California, USADivision of Oral Biology and Medicine, The Jane and Jerry Weintraub Center for Reconstructive Biotechnology, University of California Los Angeles School of Dentistry, Los Angeles, California, USADivision of Oral Biology and Medicine, The Jane and Jerry Weintraub Center for Reconstructive Biotechnology, University of California Los Angeles School of Dentistry, Los Angeles, California, USADivision of Oral Biology and Medicine, The Jane and Jerry Weintraub Center for Reconstructive Biotechnology, University of California Los Angeles School of Dentistry, Los Angeles, California, USADepartment of Biotechnology, Jožef Stefan Institute, Ljubljana, SloveniaSemel Institution, University of California Los Angeles, Los Angeles, California, USADivision of Oral Biology and Medicine, The Jane and Jerry Weintraub Center for Reconstructive Biotechnology, University of California Los Angeles School of Dentistry, Los Angeles, California, USASemel Institution, University of California Los Angeles, Los Angeles, California, USASemel Institution, University of California Los Angeles, Los Angeles, California, USADepartment of Biotechnology, Jožef Stefan Institute, Ljubljana, SloveniaObjective Natural killer (NK) cells are the largest innate lymphocyte subset with potent antitumour and antiviral functions. However, clinical utilisation of human NK cells is hampered due to a lack of reliable methods to augment their antitumour potential. We demonstrated technology in which human NK cells were cocultured with osteoclasts in the presence of probiotic bacteria. This approach significantly augmented the antitumour cytotoxicity and polyfunctionality of human NK cells, resulting in the generation of supercharged NK (sNK) cells.Methods and analysis We explored the proteomic, transcriptomic and functional characterisation of sNK cells using cell imaging, flow cytometric analysis, 51-chromium release cytotoxicity assay, ELISA, ELIspot, IsoPLexis single-cell secretome analysis, proteomic analysis, RNA analysis, western blot and enzyme kinetics.Results We found that sNK cells were less susceptible to split anergy and tumour-induced exhaustion. Proteomic analyses revealed that sNK cells significantly increased their cell motility and proliferation. Single-cell transcriptomes uncovered sNK cells undertaking a unique differentiation trajectory and turning on STAT1, JUN, BHLHE40, ELF1, MAX and MYC regulons essential for augmenting antitumour effector functions and proliferation, respectively. Both proteomic and single-cell transcriptomes revealed that an increase in Cathepsin C helped to augment the quantity and function of Granzyme B.Conclusions These results support that this unique method produces potent NK cells for clinical utilisation and delineate the molecular mechanisms associated with this process.https://bmjoncology.bmj.com/content/4/1/e000676.full |
| spellingShingle | Po-Chun Chen Subramaniam Malarkannan Ao Mei Kawaljit Kaur Anahid Jewett Meng-Wei Ko Emanuela Senjor Milica Perišić Nanut Lucy Wanrong Gao Paul Wong Whitaker Cohn Julian P Whitelegge Janko Kos Osteoclast-expanded supercharged NK cells perform superior antitumour effector functions BMJ Oncology |
| title | Osteoclast-expanded supercharged NK cells perform superior antitumour effector functions |
| title_full | Osteoclast-expanded supercharged NK cells perform superior antitumour effector functions |
| title_fullStr | Osteoclast-expanded supercharged NK cells perform superior antitumour effector functions |
| title_full_unstemmed | Osteoclast-expanded supercharged NK cells perform superior antitumour effector functions |
| title_short | Osteoclast-expanded supercharged NK cells perform superior antitumour effector functions |
| title_sort | osteoclast expanded supercharged nk cells perform superior antitumour effector functions |
| url | https://bmjoncology.bmj.com/content/4/1/e000676.full |
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