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RNA squencing of human NK cells activated with IL-15, a combination of IL-12, IL-15, and IL-18, or with a trimeric fusion protein 18/12/TxM

ABSTRACT: RNA squencing of human NK cells activated with IL-15, a combination of IL-12, IL-15, and IL-18, or with a trimeric fusion protein 18/12/TxM

PROVIDER: PRJNA699884 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13642627_1.fastq.gz	Fastqsanger.gz
	SRR13642627_2.fastq.gz	Fastqsanger.gz
	SRR13642628_1.fastq.gz	Fastqsanger.gz
	SRR13642628_2.fastq.gz	Fastqsanger.gz
	SRR13642629_1.fastq.gz	Fastqsanger.gz

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RNA squencing of human NK cells activated with IL-15, a combination of IL-12, IL-15, and IL-18, or with a trimeric fusion protein 18/12/TxM

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RNA squencing of human NK cells activated with IL-15 or IL-12, IL-15, and IL-18

Project description:NK cells are an emerging cancer cellular therapy and potent mediators of anti-tumor immunity. Cytokine-induced memory-like (ML) NK cellular therapy is safe and induces remissions in acute myeloid leukemia (AML) patients. However, the dynamic molecular changes that occur after memory-like differentiation in vitro are unclear. Here, control or ML NK cells purified from normal donor PBMC were generated in vitro. Briefly, RosetteSep-purified NK cells were incubated in IL-12, IL-15, and IL-18, or low-dose IL-15 as a control for 16-18 hours. Control or cytokine-activated NK cells were washed three times and cultured for 6 days in low-dose IL-15, which is required for NK cell survival. After 6 days, RNA was isolated from control and memory-like (ML) NK cells (IL12/15/18 activation) and RNA-sequencing performed. Because the transcription factor GATA-3 was increased specifically in ML NK cells, we hypothesized ML NK cells would exhibit a GATA-3 gene signature compared to control NK cells. Indeed, using GSEA, a significant gene signature was associated with ML NK cell differentiation. These data support the role for GATA-3 in regulating the ML NK cell molecular program.

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Transcriptional changes in IL-2, IL-12 and IL-18 stimulated human natural killer cells

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Glucocorticoids and the cytokines IL-12, IL-15 and IL-18 present in the tumor microenvironment induce PD-1 expression on human Natural Killer cells

Project description:Background: PD-1 immune checkpoint blockade has provided significant clinical efficacy across various types of cancer by unleashing both T- and NK cell-mediated anti-tumor responses. However, resistance to immunotherapy occurs for many patients, rendering the identification of the mechanisms that control PD-1 expression extremely important to increase the response to the therapy Objective: To identify the stimuli and the molecular mechanisms that induce the de novo PD-1 expression on human NK cells in the tumor setting Methods: NK cells freshly isolated from peripheral blood of healthy donors were stimulated with different combinations of molecules, and PD-1 expression was studied at the mRNA and protein level. Moreover, ex vivo analysis of tumor microenvironment and NK cell phenotype was performed. Results: Glucocorticoids (GCs) are indispensable for PD-1 induction on human NK cells, in cooperation with a combination of cytokines that are abundant at the tumor site. Mechanistically, GCs together with IL-12, IL-15 and IL-18 not only upregulate PDCD1 transcription, but also activate a previously unrecognized transcriptional program leading to enhanced mRNA translation and resulting in an increased PD-1 protein amount in NK cells. Conclusion: Our results provide evidence of a novel immune suppressive mechanism of GCs involving the transcriptional and translational control of an important immune checkpoint

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RNAseq analysis of IL-18 WT- and IL-18 D69A-overexpressing B16-F10 tumors

Project description:To gain insights into the function and mechanism of Interleukin (IL)-18 in cancer cells, we conducted RNA-seq analysis to compare the gene expression profiles between IL-18 WT- and IL-18 D69A (IL-18 mutagenesis replacing aspartate (D) with alanine (A) at position 69)-overexpressing B16-F10 tumors. As expected, many genes were upregulated in IL-18 WT samples. Notably, transcripts related to many signaling pathways such as interferon signaling pathway were significantly enriched in samples expressing IL-18 WT.

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