20.500.12556/DiRROS-22873 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 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. antitumour function protitumorska funkcija true false false BMJ Publishing Group Združeno kraljestvo Angleški jezik Slovenski jezik © Author(s) (or their employer(s)) 2025. Neznano 2025-07-07 13:36:48 2025-07-07 13:36:48 2025-10-24 03:52:24 0000-00-00 00:00:00 2025 0 0 Nasl. z nasl. zaslona; Soavtorji iz Slovenije: Emanuela Senjor, Milica Perišić Nanut, Janko Kos; Opis vira z dne 7. 7. 2025; str. 1-20 iss. 1 Vol. 4 Jan. 2025 0000-00-00 Zaloznikova Objavljeno NiDoloceno 2025-01-29 2025-04-28 2025-06-10 616-097 2752-7948 10.1136/bmjonc-2024-000676 241621763 https://bmjoncology.bmj.com/content/4/1/e000676 1 ac14418a-5b26-11f0-8080-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=33290 RAZ_Ko_Meng-Wei_2025.pdf RAZ_Ko_Meng-Wei_2025.pdf 1 C8F809BFA1E4520674D9CD9F937262D2 62622b93bee9bc69f4e2e7f06077752d0110769061ac842036767e262d457392 a7f45e2d-5b27-11f0-8080-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=33291 Institut Jožef Stefan 0 0 0