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MiRNA profiling of pooled serum circulating cell-free miRNA of Tn+ tumor-bearing mice

ABSTRACT: To investigate the systemic impact of Tn+ tumor phenotypes, we conducted an analysis of circulating miRNAs in the serum of mice bearing Tn+- 4T1 tumors. The 4T1/Tn+ cell line was generated via CRISPR/Cas9 targeting of Cosmc, and tumors were established orthotopically in BALB/c mice. Tumor-bearing animals were monitored for primary growth and micrometastatic dissemination to the lungs, followed by serum collection at endpoint. Serum samples were pooled according to tumor Tn status (Tn+ or Tn–) for total RNA extraction, including small RNAs. Using the NanoString nCounter Mouse v1.5 miRNA assay, we profiled miRNA expression in three serum pools (two Tn+, one Tn–), followed by normalization and quality control via the nCounter Advanced Analysis Software. Expression values were normalized using geometric means of internal positive and negative controls, and only stable miRNA regions (CV < 15%) were retained for downstream analysis.

ORGANISM(S): Mus musculus

PROVIDER: GSE302365 | GEO | 2026/06/23

REPOSITORIES: GEO

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Project description:Myeloid-derived suppressor cells (MDSCs) are immature myeloid cells that accumulate in the tumor microenvironment of most cancer patients. There MDSCs suppress both adaptive and innate immune responses, hindering immunotherapies. Moreover, many cancers are accompanied by inflammation, a processes that further intensifies MDSC suppressive activity, causing aggressive tumor progression and metastasis. MDSCs collected from tumor-bearing mice profusely release nano-scale membrane-bound extracellular vesicles, called exosomes, which carry biologically active proteins between cells and contribute directly to the immune suppressive functions of MDSC. Many studies on other cell types have shown that exosomes may also carry microRNAs (miRNAs) and messenger RNAs (mRNAs) which can also be transferred to surrounding and distant cells. However, to the best of our knowledge, the miRNA and mRNA cargo of MDSC-derived exosomes has not yet been interrogated. This study aims to identify and quantify the cargo of MDSC and their immunosuppressive exosomes to gather knowledge that can offer insights on the mechanisms by which MDSCs contribute to immune suppression, focusing on the role of exosomes as intercellular communication mediators in the tumor microenvironment. In order to achieve our objective a well-established mouse model based on a conventional mammary carcinoma (4T1 cells) and heightened inflammation (4T1 transduced to express the cytokine interleukin-1b) was used. We provide evidence that MDSC-derived exosomes carry proteins, mRNAs and miRNAs. Relative quantitation demonstrated quantitative differences between the exosome cargo and the cargo of their parental cells, supporting the hypothesis that selective loading into the exosomes is possible. Additionally, quantitative and functional analyses of the exosome cargo generated under conventional and heightened inflammation conditions are consistent with clinical observations that inflammation is linked to cancer development.

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MiRNA profiling of pooled serum circulating cell-free miRNA of Tn+ tumor-bearing mice

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