Adaptation of NK-92 cells to serum free culture conditions
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ABSTRACT:
PROVIDER: PRJEB33549 | ENA |
REPOSITORIES: ENA
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Project description:Identification of the mechanisms through which BET inhibitor (OTX-015) stimulates natural killer (NK) activation. RNA-seq was performed comparing vehicle- (DMSO) to OTX-015-treated NK-92 cell line.
2024-01-26 | E-MTAB-12853 | biostudies-arrayexpress
Project description:To investigate how the overexpression of a constitutively active human NHE1 may affect cytotoxicity-related gene expression in NK-92 cells, we established NK-92 cell lines stably express the NHE1 or empty vector (EV) control by lentiviral transduction.
2022-06-02 | GSE205146 | GEO
Project description:NK-92 cells and their CAR-modified derivatives exhibit strong cytotoxic activity against tumors and are promising "off-the-shelf" therapeutics compared to CAR-T cells. In order to ensure safety and prevent the occurrence of secondary tumors, (CAR-)NK-92 cells need to be inactivated before transfusion. With increasing clinical use, there is a growing demand for enhanced safety and an efficient method that stops cell proliferation, but better maintains the cytotoxic effector functions of the cells compared to commonly used gamma irradiation. Recently, low-energy electron irradiation (LEEI) was successfully applied for inactivation of bacteria and viruses for vaccine production, and was described as a method for NK-92 inactivation for the first time. In the present publication, data on extensive characterization of LEEI and its comparison with gamma irradiation for the inactivation of parental NK-92 cells as well as CD123-directed CAR-NK-92 cells are provided. Our results show that both irradiation methods cause a progressive decrease in cell viability and are therefore suitable for inhibition of proliferation. Notably, cytotoxicity of the NK cells was significantly reduced by gamma irradiation, but not by LEEI three days after irradiation, compared to non-irradiated cells. Both gamma irradiation as well as LEEI led to substantial DNA-damage and an accumulation of irradiated cells in the G2/M phase. In addition, transcriptomic analysis of cells irradiated with both methods revealed approximately 12-fold more differentially expressed genes after gamma irradiation compared to LEEI. Analysis of surface molecules revealed an irradiation-induced decrease in CD56 expression but no changes in the levels of the activating receptors NKp46, NKG2D, and NKp30. Conclusions: The present data show that LEEI efficiently inactivates (CAR )NK-92 cells and sustains their cytotoxic capacity better than gamma irradiation. Taking into account additional logistic advantages of LEEI proves a potent alternative in the manufacturing of (CAR-)NK-92 cells for clinical application.
2021-05-17 | GSE166976 | GEO
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Project description:We engrafted empty vector, wild type CCL22, and Pro79Arg-CCL22 mutant-expressing NK-92 cells into NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(IL15)1Sz/SzJ mice to assess in vivo function of detected CCL22 mutations. Engrafted Pro79Arg NK-92 cells recapitulated the phenotype of CLPD-NK patients with CCL22 mutations.
2022-01-19 | GSE163864 | GEO
Project description:Transcriptional profiling of mbIL2 activated NK-92 cells compared to recombinant human IL2 activated NK-92 cells.
2022-10-21 | GSE199391 | GEO
Project description:In this study we have compared the proteomic profile of extracellular vesicles (EVs) prepared from primary, human NK cells or the human NK cell lines NK-92 and KHYG-1 cultured for 48hrs in serum-free conditions. EVs were harvested from cells either under resting conditions (culture in IL-15) or upon activation (combination of IL-12, IL-15, and IL-18). In addition, primary NK cells were activated in the presence of anti-CD16-coated beads, and EVs harvested after 48hrs. The aim was to compare their ability to target and kill a variety of tumor cell line-derived spheroids
2022-10-14 | PXD027284 | Pride
Project description:Purpose: This study aims to evaluate the effect of FACT complex inhibitor CBL0137 to the total transcriptome of NK-92 cells through RNA-seq assay. Methods: NK-92 cells were treated with CBL0137 (500 nM) or DMSO for 24 h. RNA samples were extracted and submitted to GENEWIZ (South Plainfield, NJ 07080) for quality control and subsequent rRNA removal through polyA selection, followed by the library preparation. Sequencing was carried out on an Illumina HiSeq platform with the configuration of 2 × 150 bp (paired end), and > 20 million reads per sample were achieved. Differential expression of genes was analyzed by DESeq2. Results: R packages, pheatmap and clusterProfiler, were used for heatmap construction and pathway analysis, respectively. 917 genes were enriched based on the fold-change > 1.5 as well as the adjusted P-value < 0.05. GO biological process (BP) analysis shows that inteferon signaling and anti-viral responses are among top enriched pathways. Expression of interferons and interferon-stimulated genes (ISGs) are increased based on heatmap, which was further validated through quantitative reverse transcription PCR (RT-qPCR) assay. Conclusions: CBL0137 causes the systemic upregulation of IFNs and ISGs with statistical significance in NK-92 cells, which shows the potential of CBL0137 to trigger the anti-viral innate immunity. In addition, due to the upragulation of IFN-gamma pathways, CBL0137 also shows the ability to increase the NK cytotoxicity.
2022-07-29 | GSE184784 | GEO
Project description:Natural killer (NK) cells are a type of innate lymphocytes that play key roles in immune surveillance against tumors and viral infection. NK cells distinguish abnormal cells from healthy cells by cell-cell interaction with cell surface proteins and then attack target cells via multiple mechanisms involving TRAIL, Fas Ligand, cytokine secretion, perforin, and granzymes. In addition, extracellular vesicles (EVs), including exosomes derived from NK cells (NK-EVs), possess cytotoxic capacity against tumor cells, but their characteristics and regulation by cytokines remain unknown. Here, we report that EVs derived from human NK-92 cells stimulated with IL-15 + IL-21 show enhanced cytotoxic capacity against tumor cells in a granzyme B independent manner. In addition, small RNA-seq and mass spectrometry analyses indicate that miRNA and protein profiles in EVs are altered by cytokine stimulation. We also show NK-EVs are taken up by target cells via macropinocytosis. Collectively, our findings reveal novel characteristics of NK-EVs and the mechanism of their incorporation into target cells.
2021-09-09 | PXD018982 | Pride