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Next Generation Sequencing Facilitates Quantitative Analysis of Renca and B16 Murine Cancer Line Transcriptomes

ABSTRACT: Renca, a murine renal cell carcinoma, and B16F10, a murine melanoma, were transcriptionally profiled in a study designed to accompany an in vivo CRISPR screen to identify novel immunotherapy dependencies.

ORGANISM(S): Mus musculus

PROVIDER: GSE162105 | GEO | 2020/11/25

REPOSITORIES: GEO

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Next Generation Sequencing Facilitates Quantitative Analysis of Renca and B16 Murine Cancer Line Transcriptomes

Project description:Next Generation Sequencing Facilitates Quantitative Analysis of Renca and B16 Murine Cancer Line Transcriptomes

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Comparison of Renca tumor cells growing intrakidney or subcutaneously

Project description:Tumors can occur in many sites in the body, but how tissues at different anatomical sites affect tumor microenvironments and their subsequent response to therapy is not known. Here we show that different tissues can cause the same tumor to develop fundamentally different microenvironments resulting in dramatic changes in responses to immunotherapy. Whereas established subcutaneous tumors could be eradicated in the majority of mice using a combination of three agonist antibodies, tumors in visceral organs responded much less. Orthotopic kidney tumors had a microenvironment with a higher frequency of alternatively-activated macrophages and their associated molecules. Neutralizing the CCL2 chemokine or IL-13 cytokine, important in macrophage biology led to a greatly improved therapeutic effect. Importantly, a significantly reduced therapeutic response of subcutaneous tumors was observed when visceral tumors were simultaneously present in the same mice. In this study we sought to compare gene expression of mouse renal cell carcinoma cells growing in the kidney to cells growing subcutaneously. Renca is a renal cell carcinoma cell line (Murphy and Hrushesky, 1973). To produce subcutaneous tumors, mice received 2 x 10^5 Renca. Orthotopic tumors were established by first surgically isolating the organ and then injecting 2 x 10^5 Renca cells intrakidney (IK) into the outer cortex of the kidney in 20 ul of PBS. Tumors were removed from mice 12 days after implantation, when approximately 20-30 mm2. Tumors were snap frozen on liquid nitrogen prior to processing for total RNA followed by analysis on Mouse Gene 1.0 chips. Kidney Renca tumors were excised from 3 BALB/c mice and subcutaneous Renca tumors were excised from another 3 BALB/c mice. cDNA was prepared and hybridized to Mouse Gene 1.0 ST chips.

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Gene expression profiling of murine tumors derived from Renca control knockout vs Pbrm1 knockout tumor clones

Project description:We investigated the influence of Pbrm1 on the tumor microenvironment in murine tumors. We employed a Renca-BALB/c murine immune competent model for our study. We compared Renca control knockout tumors to two Pbrm1 KO tumor clones (n=5 for each of two clones).

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Comparison of Renca tumor cells growing intrakidney or subcutaneously

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Gene expression changes in mouse kidney cancer cells (RENCA) after anti-PD-L1 resistance

Project description:RNA sequencing analysis was used to identify changes in mouse kidney cancer cells (RENCA) following resistance to anti-PD-L1 (PDR).

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CXCR4 inhibition modulates the tumor microenvironment and retards the growth of B16-OVA melanoma and Renca tumors.

Project description:To determine whether blockade of the chemokine receptor CXCR4 might alter the tumor microenvironment and inhibit tumor growth, we tested the efficacy of the CXCR4 antagonist X4-136 as a single agent and in combination with various immune checkpoint inhibitors in the syngeneic murine melanoma model B16-OVA. We also tested its activity alone and in combination with axitinib in the renal cancer model Renca. We found that X4-136 exhibited potent single agent antitumor activity in the B16-OVA model that was additive to that of an anti-PDL1 antibody. The antitumor activities were associated with a reduction in the number of immunosuppressive regulatory T cells and myeloid-derived suppressor cells and an increase in the number of tumor-specific CD8/perforin cells in the tumor-microenvironment. Apart from these immune effects, X4-136 alone and in combination with checkpoint inhibitors inhibited the Akt/FOXO-3a cell survival pathway in vitro and in vivo, suggesting that it might have antitumor activity independent of its effects on immune cell trafficking. Similar effects on tumor growth and cytotoxic T lymphocytes infiltration were observed in the Renca model. These studies show that the effects of CXCR4 blockade on immune cell trafficking might serve as a useful adjunct to immune checkpoint inhibitors and other therapies in the treatment of cancer.

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Gene expression changes in mouse kidney cancer cells (RENCA) after sunitinib resistance and acute and chronic withdrawal

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The effect of TIM-3 stimulation on the gene expression profile of B16-F10 melanoma cells

Project description:The effect of TIM-3 stimulation was studied on B16F10 mouse melanoma cells, in vitro.

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