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Cancer-educated mesenchymal stem cells promote pre-metastatic niche formation for disseminated cancer cells via the induction of PMN-MDSC expansion

ABSTRACT: We subcutaneously injected the RFP-LLC cells and GFP-BMSCs into C57BL/6 mice and then collected GFP-BMSCs from the primary inoculation cancer site and from the bone marrow by flow cytometric sorting technology.

ORGANISM(S): Mus musculus

PROVIDER: GSE120456 | GEO | 2019/09/01

REPOSITORIES: GEO

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RNA sequencing in LLCs from co-injected mice or LLC injection alone.

Project description:we analyzed transcriptomic profiles in LLC cells isolated from primary cancer sites. C57BL/6 mice were subcutaneously injected with LLC-RFP with or without BMSCs.

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Stroke Exacerbates Cancer Progression by Upregulating LCN2 in PMN-MDSC.

Project description:Acute ischemic stroke (AIS) is common in patients with cancer, and mounting clinical evidence suggests that it may shorten the survival of cancer patients. But how stroke affects the progression of cancer remains unclear. We inoculated B16 tumor cells (2 × 105) subcutaneously before distal middle cerebral artery occlusion (dMCAO) or sham surgery in C57BL/6 mice and found that compared to sham operated mice, dMCAO mice developed significantly increased tumor volume and were accompanied by lower survival rate. To explore the underlying mechanism, we performed RNA-sequencing analysis of the tumor tissue from mice with or without stroke and found prominent upregulation of lipocalin 2 (LCN2) in the tumor from stroke mice compared to those from sham mice. Using quantitative reverse transcription-PCR, we confirmed increased mRNA expression of LCN2 as well as anti-inflammatory cytokines-Arg1, IL-10, and decreased mRNA level of pro-inflammatory cytokines-IL-6, IL-23 in the tumor of cancer-bearing stroke mice. Both immunofluorescence staining and flow cytometry analysis revealed that increased expression of LCN2 was mainly derived from the polymorphonuclear myeloid derived suppressor cells (PMN-MDSCs) in the tumor. We also found that stroke reduced the PMN-MDSCs in the peripheral blood, but increased PMN-MDSCs in the tumor of the cancer-bearing mice after stroke. In conclusion, cerebral ischemic stroke may exacerbate cancer progression by increasing LCN2 expression in PMN-MDSCs, which turns out to be a promising therapeutic target to suppress cancer progression after ischemic stroke.

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Gene expression profiling of mature dendritic cells from mice s. c. treated with GM-CSF-gene transduced LLC cells (LLC/SeV/GM) or control cells (LLC, LLC/SeV/GFP)

Project description:Irradiated granulocyte macrophage-colony stimulating factor (GM-CSF)-transduced autologous tumor cells induce substantial antitumor immunity through the maturation and migration of dendritic cells (DCs). However, little is known about the key molecules involved in GM-CSF-sensitized DCs (GM-DCs) in tumor draining lymph nodes (TDLNs). We initially confirmed that mice subcutaneously injected with poorly immunogenic syngeneic Lewis lung carcinoma (LLC) cells transduced with Sendai virus encoding GM-CSF (LLC/SeV/GM) significantly rejected the tumor growth. Using microarray expression profiling, we obtained a large number of gene expression data files from GM-DCs and control DCs in TDLNs, and subjected them to network-based cluster analysis and unexpectedly unraveled the expression levels of type I IFNs-related genes specifically expressed in plasmacytoid DCs (pDC) were robustly up-regulated in GM-DCs. In vivo depletion assay showed that pDC-depleted mice treated with subcutaneous LLC/SeV/GM cells abrogated the antitumor effects observed in control mice. Moreover combination use of imiquimod for TLR7 triggering on pDC with irradiated LLC/SeV/GM cells induced a significant therapeutic antitumor effect with marked induction of CD9+ pDC with antitumor phenotype, whereas other control mice groups had only minimal to-modest antitumor responses, implicating that this combined vaccine strategy using imiquimod could be promising for improvement of GM-CSF-induced antitumor immunity. Mouse GM-CSF induced gene expression in mature dendritic cells in tumor draining lymph nodes from C57/BL6N female mouse was measured at 2 days after s.c. tumor challenge with GM-CSF gene-transduced LLC cells (LLC/SeV/GM) or control cells (LLC, LLC/SeV/GFP).

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Transcription profiling by array of mouse E15.5 FACS sorted pancreas populations based on double fluorescent reporter (Hes1GFP and Neurog3-RFP), DBA staining and Ptf1a-YFP.

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Endothelial-to-osteoblast transition in normal mouse bone development and postnatal growth

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Upregulated TRAIL and Reduced DcR2 Mediate Apoptosis of Decidual PMN-MDSC in Unexplained Recurrent Pregnancy Loss.

Project description:Myeloid-derived suppressor cells (MDSC), especially polymorphonuclear MDSC (PMN-MDSC), accumulate in maternal-fetal interface during pregnancy and are involved in the maintenance of immune tolerance. Decreased PMN-MDSC is associated with pregnancy complications such as unexplained recurrent pregnancy loss (URPL). In the present study we showed decreased PMN-MDSC in the URPL group compared with the normal pregnancy (NP) group, and PMN-MDSC was the major subset of MDSC in human decidua with potent immune suppression activity. We then performed gene expression profile and found that human decidual PMN-MDSC in the NP and URPL groups showed different gene and pathway signature, including apoptosis. Apoptosis of decidual PMN-MDSC was mediated by TNF-related apoptosis-induced ligand (TRAIL) in a Caspase 3 dependent manner. TRAIL was expressed in decidua and upregulated in decidua of the URPL group. Notably, of all the membrane TRAIL receptors, only DcR2 was down-regulated in PMN-MDSC in the URPL group. In vitro experiment demonstrated that DcR2 blockade sensitized PMN-MDSC to TRAIL-mediated apoptosis. Together, these data indicate that increased TRAIL and reduced DcR2 on PMN-MDSC sensitize PMN-MDSC response to TRAIL-induced apoptosis in the URPL group, which is responsible for decreased accumulation of PMN-MDSC in URPL.

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Tumor-specific mechanism of Treg enrichment mediated by the integrin avb8

Project description:Subcutaneous tumors from mock transfected LLC or b8 transfected LLC were established on flanks of C57B/6 FoxP3-Ires GFP mice. Mice were treated with isotype control antibodies or anti-b8 and Tregs (CD4+ GFP+) were sorted, pooled and total RNA isolated for RNAseq.

2021-02-23 | GSE158031 | GEO

Genome-wide reporter screen for transcriptional regulators of ribosome biogenesis genes and ribosomal protein genes

Project description:A genome-wide CRISPR/Cas9 knockout reporter screen was performed to identify transcriptional regulators of ribosomal biogenesis genes and ribosomal protein genes. The murine GeCKO v2 library provided by the Zhang lab was amplified and positive control sgRNAs targeting the reporters were spiked-in. NIH/3T3 cells (murine fibroblasts) harboring the "ribosome biogenesis" reporter Fbl-GFP (Fibrillarin promoter-driven enhanced GFP fused to the PEST domain of the murine ornithine decarboxylase) and the "ribosomal protein" reporter Rpl18-RFP (Rpl18 promoter-driven TurboRFP fused to the PEST domain of the murine ornithine decarboxylase) were transduced with the aforementioned library, selected and FACS-sorted six days after infection. The sorted conditions were i) low GFP, but middle to strong RFP expression (RiBi down); ii) low RFP, but middle to strong GFP expression (RP down); iii) low GFP and low RFP expression (Both down); iv) strong GFP and strong RFP expression (Both up). The same amount of cells that were used for sorting, were harvested as the "unsorted" condition to which all sorted samples were compared to. The plasmid library was also sequenced.

2022-07-13 | GSE207900 | GEO

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