A.F. Biggi , R.N. Silvestre , M.C. Tirapelle , J.T. de Azevedo , D. Covas , K.C. Malmegrim , M.L. Figueiredo , V. Picanço-Castro
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引用次数: 0
Abstract
Background & Aim
Interleukin 27 (IL-27), part of the IL-12 cytokine family, enhances NK cell cytotoxicity by inducing activating receptors (e.g., NKp46, NKG2D), and stimulating IFN-gamma production. While chimeric antigen receptor (CAR) has shown promise for NK cells in early-phase clinical trials, improving CAR-NK cell therapeutic effectiveness is critical. Although transgenic expression of cytokines (e.g., IL-15) in CAR-NK cells partially enhances pro-survival or proliferative properties, further research is needed to identify the optimal cytokine. Therefore, investigating the role of co-expressing IL-27 with a fourth-generation CD19-targeted CAR on NK cells is crucial.
Methods, Results & Conclusion
Lentiviral particles carrying CAR19 and CAR19-IL27 vectors were used to transduce NK-92 cell line. CAR+ cells were FACS-sorted and the potency of CAR-NK cells was evaluated against CD19+ cell lines (NALM-6 and Raji) both in vitro and in in vivo murine models. Tumor burden was quantified through bioluminescence. RNA from target-activated CAR19 and CAR19-IL27 was prepared following Illumina's TruSeq-stranded-mRNA protocol and conducted by LC Sciences. Our findings suggest that CAR19-IL27 enhances NK-92 inherent proliferative ability by about 4-fold compared to CAR19. Furthermore, CAR19-IL27 cells exhibited higher in vitro cytotoxicity against NALM-6 and Raji compared to CAR19, p<0.005 (NALM-6) and p<0.00005 (Raji); higher activation-induced degranulation (CD107a MFI), and IFN-gamma secretion; and decreased expression of inhibitory checkpoint CTLA-4 at both protein (-20-fold) and RNA levels (-1,45 log2FC). Potentially, IL-27 enhances NK cell cytotoxicity through MAPK-related pathways, as genes associated with this pathway were upregulated in CAR19-IL27 cells (e.g., CCR7, ICAM-1, IGF2, NTRK2, and NTRK3). Furthermore, in vivo experiments show that CAR19-IL27 NK cells, in contrast to CAR19 cells, exhibit an extended ability to control tumor growth, reducing tumor burden on day 21 (4 mice/group). Therefore, integrating IL-27 into CAR-NK cells is a promising strategy to boost therapeutic responses against cancer cells, supporting the evidence of fourth-generation CAR engineering in advancing NK cell-based immunotherapies. Financially supported by FAPESP 2020/07055-9, 2014/50947-7, 2013/08135-2; CNPq 440543/2022-3; CAPES 88887.817043/2023-00.
期刊介绍:
The journal brings readers the latest developments in the fast moving field of cellular therapy in man. This includes cell therapy for cancer, immune disorders, inherited diseases, tissue repair and regenerative medicine. The journal covers the science, translational development and treatment with variety of cell types including hematopoietic stem cells, immune cells (dendritic cells, NK, cells, T cells, antigen presenting cells) mesenchymal stromal cells, adipose cells, nerve, muscle, vascular and endothelial cells, and induced pluripotential stem cells. We also welcome manuscripts on subcellular derivatives such as exosomes. A specific focus is on translational research that brings cell therapy to the clinic. Cytotherapy publishes original papers, reviews, position papers editorials, commentaries and letters to the editor. We welcome "Protocols in Cytotherapy" bringing standard operating procedure for production specific cell types for clinical use within the reach of the readership.