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Gene expression profile at single cell level of FACS sorted CD8+ T cells from subcutaneous MC38 tumors from WT and Susd2-/- mice

ABSTRACT: Gene expression profile at single cell level of FACS sorted CD8+ T cells from subcutaneous MC38 tumors from WT and Susd2-/- mice

PROVIDER: PRJNA867183 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR20958500_1.fastq.gz	Fastqsanger.gz
	SRR20958500_2.fastq.gz	Fastqsanger.gz
	SRR20958501_1.fastq.gz	Fastqsanger.gz
	SRR20958501_2.fastq.gz	Fastqsanger.gz
	SRR20958502_1.fastq.gz	Fastqsanger.gz

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Gene expression profile at single cell level of FACS sorted CD8+ T cells from subcutaneous MC38 tumors from WT and Susd2-/- mice

Project description:Tumor-infiltrating CD8+ T cells are phenotypically heterogeneous with varying degrees of functional defects. We used single cell RNA sequencing (scRNA-seq) to analyze their sex differences.

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Project description:To determine differential metabolic gene expression in distinct cell populations from subcutaneous MC38 tumors at baseline and in response to rapamycin, tumors from mice treated with vehicle or rapamycin were harvested, reconstituted to single cell suspensions, and flow sorted into cancer cell (CD45-), tumor-associated macrophage (TAM, CD45+ CD11b+ Ly6G- Ly6C lo F4/80 hi), monocytic myeloid-derived suppressor cell (M-MDSC, CD45+ CD11b+ Ly6G- Ly6C hi), CD8 T cell (CD45+ CD3+ CD8a+), and CD4 T cell (CD45+ CD3+ CD8a-) populations. RNA was extracted and transcripts were quantified by the NanoString nCounter Metabolic Pathways Panel.

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Effect of rapamycin on murine subcutaneous MC38 tumor cell populations

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Transcriptional profiling of IL6R-deficient CD8 T cells isolated from subcutaneous MC38 tumors treated with anti-PD-L1 therapy

Project description:Interleukin 6 (IL-6) is a pleiotropic cytokine with diverse roles in homeostasis, inflammation, and cancer. In multiple syngeneic mouse tumor models, we found that blockade of IL-6 signaling (using an IL6R-blocking antibody) synergized with anti-PD-L1 therapy to drive potent anti-tumor CD8 T cell responses and tumor rejection. To better characterize the cell-intrinsic effects of IL-6 signaling in tumor-reactive CD8+ T cells during anti-PD-L1 therapy, we generated mice with genetic IL6R deficiency restricted to CD8 T cells by crossing IL6R.loxp and E8i.CD8.Cre mice (CD8ΔIL6R mice). Compared to WT littermate controls, we found that CD8ΔIL6R mice had stronger respones to anti-PD-L1 therapy in terms of improved CD8 T cell function (e.g. increased production of IFNγ and TNF, measured by flow cytometry) and enhanced tumor control, suggesting that direct IL-6 signaling in CD8 T cells is sufficient to impair anti-tumor immunity. In this study we aimed to characterize the phenotype of IL6R-deficient CD8 T cells in more detail via whole-transcriptome profiling. CD8ΔIL6R and WT littermates were implanted with MC38 tumors in the right flank; when tumors reached ~150mm3 in volume, animals were randomized to isotype control or anti-PD-L1 treatment. CD8 T cells were FACS-purified from tumor tissue 7 days later and profiled by bulk RNAseq. Compared to cells from WT mice, CD8 T cells from CD8ΔIL6R mice showed increased expression of interferon-driven gene signatures, increased expression of cell cycle genes, and increased expression of genes critical for oxidative phosphorylation. In contrast, WT cells had higher expression of genes associated with naive and memory precursor cells. Thus, IL-6 signaling in tumor-reactive CD8 T cells limits their capacity to differentiate into potent anti-tumor effectors.

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