Human lung cancer neutrophils generate NETs with preserved anti-tumor cytotoxicity but impaired anti-migratory activity
Neutrophil extracellular traps (NETs) are DNA-protein structures released during a form of programmed neutrophil death known as NETosis. While NETs have been implicated in both tumor inhibition and promotion, their functional role in cancer remains ambiguous. In this study, we compared the NET-formi...
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Frontiers Media S.A.
2025-08-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1643609/full |
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| author | Nuha Alsharif Nuha Alsharif Mohamad Qaisi Merav Shaul Naomi Kaisar-Iluz Naomi Kaisar-Iluz Dan Padawer Dan Padawer Osnath Bouhanna Osnath Bouhanna Yael Volman Yael Volman Zvi G. Fridlender Zvi G. Fridlender |
| author_facet | Nuha Alsharif Nuha Alsharif Mohamad Qaisi Merav Shaul Naomi Kaisar-Iluz Naomi Kaisar-Iluz Dan Padawer Dan Padawer Osnath Bouhanna Osnath Bouhanna Yael Volman Yael Volman Zvi G. Fridlender Zvi G. Fridlender |
| author_sort | Nuha Alsharif |
| collection | DOAJ |
| description | Neutrophil extracellular traps (NETs) are DNA-protein structures released during a form of programmed neutrophil death known as NETosis. While NETs have been implicated in both tumor inhibition and promotion, their functional role in cancer remains ambiguous. In this study, we compared the NET-forming capacity and functional effects of NETs derived from lung cancer (LC) patients and healthy donors (H). Neutrophils were isolated from peripheral blood and stimulated in vitro with phorbol 12-myristate 13-acetate (PMA) to induce NETosis. Isolated NETs were quantified and assessed for their cytotoxicity against A549 lung cancer cells and their impact on cancer cell migration. Whereas LC neutrophils (LCN) were less cytotoxic to tumor cells than H neutrophils (HN), their NETs maintained similar tumoricidal capacity – 41.6% ± 25.3% (LCN) vs. 46.4% ± 14.5% (HN), ns. Interestingly, we noted a correlation between the amount of NETs and their cytotoxicity to tumor cells. This effect could not be recapitulated with purified genomic DNA, inducing only 3.99% of cytotoxicity to tumor cells, and confirming that intact NETs are required for the anti-tumor activity. LCN displayed an increased frequency of NETosis following PMA stimulation, yet produced significantly fewer NETs per cell – 1569 ± 306 ng (LCN) vs. 2619 ± 313 ng (HN); p = 0.025. Reactive oxygen species (ROS) production was elevated in LC neutrophils, indicating that the NETosis defect was not due to impaired oxidative burst. LCN had increased expression of immunosuppression (PDL-1) as well as exhaustion and aging markers CD62L and CD11b). Only NETs from HN inhibited the migration of A549 tumor cells, whereas those from LCN failed to suppress, and in some cases appeared to enhance, cell motility. Our data suggest that NETs in lung cancer retain anti-tumor cytotoxicity capabilities but lose their anti-migratory capacity, highlighting their dual role in tumor biology and potential as therapeutic targets. |
| format | Article |
| id | doaj-art-5d25b2e8fcae4d839811d3fa88b6f73b |
| institution | Kabale University |
| issn | 1664-3224 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Immunology |
| spelling | doaj-art-5d25b2e8fcae4d839811d3fa88b6f73b2025-08-25T12:18:11ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-08-011610.3389/fimmu.2025.16436091643609Human lung cancer neutrophils generate NETs with preserved anti-tumor cytotoxicity but impaired anti-migratory activityNuha Alsharif0Nuha Alsharif1Mohamad Qaisi2Merav Shaul3Naomi Kaisar-Iluz4Naomi Kaisar-Iluz5Dan Padawer6Dan Padawer7Osnath Bouhanna8Osnath Bouhanna9Yael Volman10Yael Volman11Zvi G. Fridlender12Zvi G. Fridlender13Institute of Pulmonary Medicine, Hadassah Hebrew University Medical Center, Jerusalem, IsraelFaculty of Medicine, Hebrew University, Jerusalem, IsraelInstitute of Pulmonary Medicine, Hadassah Hebrew University Medical Center, Jerusalem, IsraelInstitute of Pulmonary Medicine, Hadassah Hebrew University Medical Center, Jerusalem, IsraelInstitute of Pulmonary Medicine, Hadassah Hebrew University Medical Center, Jerusalem, IsraelFaculty of Medicine, Hebrew University, Jerusalem, IsraelInstitute of Pulmonary Medicine, Hadassah Hebrew University Medical Center, Jerusalem, IsraelFaculty of Medicine, Hebrew University, Jerusalem, IsraelInstitute of Pulmonary Medicine, Hadassah Hebrew University Medical Center, Jerusalem, IsraelFaculty of Medicine, Hebrew University, Jerusalem, IsraelInstitute of Pulmonary Medicine, Hadassah Hebrew University Medical Center, Jerusalem, IsraelFaculty of Medicine, Hebrew University, Jerusalem, IsraelInstitute of Pulmonary Medicine, Hadassah Hebrew University Medical Center, Jerusalem, IsraelFaculty of Medicine, Hebrew University, Jerusalem, IsraelNeutrophil extracellular traps (NETs) are DNA-protein structures released during a form of programmed neutrophil death known as NETosis. While NETs have been implicated in both tumor inhibition and promotion, their functional role in cancer remains ambiguous. In this study, we compared the NET-forming capacity and functional effects of NETs derived from lung cancer (LC) patients and healthy donors (H). Neutrophils were isolated from peripheral blood and stimulated in vitro with phorbol 12-myristate 13-acetate (PMA) to induce NETosis. Isolated NETs were quantified and assessed for their cytotoxicity against A549 lung cancer cells and their impact on cancer cell migration. Whereas LC neutrophils (LCN) were less cytotoxic to tumor cells than H neutrophils (HN), their NETs maintained similar tumoricidal capacity – 41.6% ± 25.3% (LCN) vs. 46.4% ± 14.5% (HN), ns. Interestingly, we noted a correlation between the amount of NETs and their cytotoxicity to tumor cells. This effect could not be recapitulated with purified genomic DNA, inducing only 3.99% of cytotoxicity to tumor cells, and confirming that intact NETs are required for the anti-tumor activity. LCN displayed an increased frequency of NETosis following PMA stimulation, yet produced significantly fewer NETs per cell – 1569 ± 306 ng (LCN) vs. 2619 ± 313 ng (HN); p = 0.025. Reactive oxygen species (ROS) production was elevated in LC neutrophils, indicating that the NETosis defect was not due to impaired oxidative burst. LCN had increased expression of immunosuppression (PDL-1) as well as exhaustion and aging markers CD62L and CD11b). Only NETs from HN inhibited the migration of A549 tumor cells, whereas those from LCN failed to suppress, and in some cases appeared to enhance, cell motility. Our data suggest that NETs in lung cancer retain anti-tumor cytotoxicity capabilities but lose their anti-migratory capacity, highlighting their dual role in tumor biology and potential as therapeutic targets.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1643609/fullneutrophil extracellular traps (NETs)direct-cell killingtumor microenvironmentlung cancerneutrophils |
| spellingShingle | Nuha Alsharif Nuha Alsharif Mohamad Qaisi Merav Shaul Naomi Kaisar-Iluz Naomi Kaisar-Iluz Dan Padawer Dan Padawer Osnath Bouhanna Osnath Bouhanna Yael Volman Yael Volman Zvi G. Fridlender Zvi G. Fridlender Human lung cancer neutrophils generate NETs with preserved anti-tumor cytotoxicity but impaired anti-migratory activity Frontiers in Immunology neutrophil extracellular traps (NETs) direct-cell killing tumor microenvironment lung cancer neutrophils |
| title | Human lung cancer neutrophils generate NETs with preserved anti-tumor cytotoxicity but impaired anti-migratory activity |
| title_full | Human lung cancer neutrophils generate NETs with preserved anti-tumor cytotoxicity but impaired anti-migratory activity |
| title_fullStr | Human lung cancer neutrophils generate NETs with preserved anti-tumor cytotoxicity but impaired anti-migratory activity |
| title_full_unstemmed | Human lung cancer neutrophils generate NETs with preserved anti-tumor cytotoxicity but impaired anti-migratory activity |
| title_short | Human lung cancer neutrophils generate NETs with preserved anti-tumor cytotoxicity but impaired anti-migratory activity |
| title_sort | human lung cancer neutrophils generate nets with preserved anti tumor cytotoxicity but impaired anti migratory activity |
| topic | neutrophil extracellular traps (NETs) direct-cell killing tumor microenvironment lung cancer neutrophils |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1643609/full |
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