Mus musculus
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA219401 | ENA |
REPOSITORIES: ENA
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Project description:Myeloid-derived suppressor cells (MDSCs) are one of the main cell populations that negatively regulate immune responses. However, the mechanism underlying the differentiation and expansion of MDSCs remains unclear. Employing a low-density miRNA microarray and a TaqMan probe-based qRT-PCR assay, we investigated the miRNAs expression profiles of MDSC. Compared to freshly isolated BM cells, BM-MDSCs had a significantly altered miRNA profile. There were 45 miRNAs that were upregulated and 17 miRNAs that were downregulated more than 2-fold. Among these miRNAs, miR-155 and miR-21 have the highest upregulation after cytokine induction. BM cells were planted into dishes using RPMI1640 medium supplemented with 2 mM L-glutamine, 10mM HEPES, 20M-BM-5M 2-mercaptoethanol, 150 U/ml streptomycin, 200 U/ml penicillin, 10% FBS and stimulated with combinations of GM-CSF (40ng/ml), and IL-6 (40ng/ml). Cells were cultured at 37M-BM-0C in 5% CO2-humidified atmosphere for 4 days.Total RNA were isolated, low-density miRNA microarray was performed
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Project description:The aim of the study is to evaluate whether the preoperative level of myeloid-derived suppressor cells is associated with postoperative complications classified by Clavien-Dindo categories. Levels of all MDSC, polymorphonuclear MDSC (PMNMDSC), monocytic MDSC (MMDSC), early-stage MDSC (EMDSC) and monocytic to polymorphonuclear MDSC ratio (M/PMN MDCS) were established and compared in patients with postoperative complications, severe postoperative complications (>= IIIA according to Clavien-Dindo) and severe septic complications.
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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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