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Systematic investigation of cytokine signaling activity at the tissue and single-cell levels

ABSTRACT: Purpose: The role of different TGF-beta isoforms in tumor progression is not well established. The goal of the study was to assess the effect on the 4T1 mouse mammary tumor transcriptomes of treatment with isoform-selective anti-TGFbeta neutralizing antibodies to address whether sparing TGF-beta3 had any biological impact. Methods: 40,000 4T1 cells were implanted in the #4 mammary fatpad of syngeneic female BALB/c mice. Mice were treated with therapeutic or control antibodies (5mg/Kg i.p. 3x q wk) for 12 days prior to tumor harvest for RNA-Seq analysis. Treatments were: 1D11 pan-TGFbeta mouse monoclonal, XPA.42.681 pan-TGFbeta human monoclonal, XPA.42.089 anti-TGFbeta1+2 human monoclonal, anti-KLH control human monoclonal, 13C4 control mouse monoclonal, or vehicle control. Tumor mRNA profiles were generated by deep-sequencing of n=5 to 7 samples/treatment group using Illumina HiSeq2500 with TruSeq v4 chemistry. Results: 51-115 million pass-filtered reads were obtained for all samples, with >=91% of bases having a quality score of 30 or above.

ORGANISM(S): Mus musculus

PROVIDER: GSE174686 | GEO | 2021/07/26

REPOSITORIES: GEO

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Project description:Accumulating data support the concept that ionizing radiation therapy (RT) has the potential to convert the tumor into an in situ, individualized vaccine; however this potential is rarely realized by RT alone. Transforming growth factor β (TGFβ) is an immunosuppressive cytokine that is activated by RT and inhibits the antigen-presenting function of dendritic cells and the differentiation of effector CD8+ T cells. Here we tested the hypothesis that TGFβ hinders the ability of RT to promote anti-tumor immunity. Development of tumor-specific immunity was examined in a pre-clinical model of metastatic breast cancer. Mice bearing established 4T1 mouse mammary carcinoma treated with pan-isoform specific TGFβ neutralizing antibody, 1D11, showed significantly improved control of the irradiated tumor and non-irradiated metastases, but no effect in the absence of RT. Notably, whole tumor transcriptional analysis demonstrated the selective upregulation of genes associated with immune-mediated rejection only in tumors of mice treated with RT+TGFβ blockade. Mice treated with RT+TGFβ blockade exhibited cross-priming of CD8+ T cells producing IFNγ in response to three tumor-specific antigens in tumor-draining lymph nodes, which was not evident for single modality treatment. Analysis of the immune infiltrate in mouse tumors showed a significant increase in CD4+ and CD8+ T cells only in mice treated with the combination of RT+TGFβ blockade. Depletion of CD4+ or CD8+ T cells abrogated the therapeutic benefit of RT+TGFβ blockade. These data identify TGFβ as a master inhibitor of the ability of RT to generate an in situ tumor vaccine, which supports testing inhibition of TGFβ during radiotherapy to promote therapeutically effective anti-tumor immunity. We used genome-wide microarray to depict main biological processes responsibles for the therapeutic benefit of the combination ofTGF-beta blockade and local radiotherapy. To gain a more comprehensice protrait of the effects of RT and TGFbeta blockade on gene expressionin tumors, we collected 4T1 tumors 4 days after completion of RT. Three tumors from each group were then subjected to RNA extraction and hybridization on affymetrix array.

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Systematic investigation of cytokine signaling activity at the tissue and single-cell levels

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