Apoptotic cells induce proliferation of peritoneal macrophages
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA691612 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR13422665.fastq.gz | Fastqsanger.gz | |||
| SRR13422666.fastq.gz | Fastqsanger.gz | |||
| SRR13422667.fastq.gz | Fastqsanger.gz | |||
| SRR13422668.fastq.gz | Fastqsanger.gz | |||
| SRR13422669.fastq.gz | Fastqsanger.gz |
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Project description:The interaction of macrophages with apoptotic cells is required for efficient resolution of inflammation. While apoptotic cell removal prevents inflammation due to secondary necrosis, it also alters the macrophage phenotype to hinder further inflammatory reactions. The interaction between apoptotic cells and macrophages is often studied by chemical or biological induction of apoptosis, which may introduce artifacts by affecting the macrophages as well and/or triggering unrelated signaling pathways. Here, we set up a pure cell death system in which NIH 3T3 cells expressing dimerizable Caspase-8 were co-cultured with peritoneal macrophages in a transwell system. Phenotype changes in macrophages induced by apoptotic cells were evaluated by RNA sequencing, which revealed an unexpectedly dominant impact on macrophage proliferation. This was confirmed in functional assays with primary peritoneal macrophages and IC-21 macrophages. Moreover, inhibition of apoptosis during Zymosan-induced peritonitis in mice decreased mRNA levels of cell cycle mediators in peritoneal macrophages. Proliferation of macrophages in response to apoptotic cells may be important to increase macrophage numbers to allow efficient clearance and resolution of inflammation.
2021-03-18 | GSE164709 | GEO
Project description:The interaction of macrophages with apoptotic cells is required for efficient resolution of inflammation. While apoptotic cell removal prevents inflammation due to secondary necrosis, it also alters the macrophage phenotype to hinder further inflammatory reactions. The interaction between apoptotic cells and macrophages is often studied by chemical or biological induction of apoptosis, which may introduce artifacts by affecting the macrophages as well and/or triggering unrelated signaling pathways. Here, we set up a pure cell death system in which NIH 3T3 cells expressing dimerizable Caspase-8 were co-cultured with peritoneal macrophages in a transwell system. Phenotype changes in macrophages induced by apoptotic cells were evaluated by RNA sequencing, which revealed an unexpectedly dominant impact on macrophage proliferation. This was confirmed in functional assays with primary peritoneal macrophages and IC-21 macrophages. Moreover, inhibition of apoptosis during Zymosan-induced peritonitis in mice decreased mRNA levels of cell cycle mediators in peritoneal macrophages. Proliferation of macrophages in response to apoptotic cells may be important to increase macrophage numbers in order to allow efficient clearance and resolution of inflammation.
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Project description:Macrophages induce inflammation by efferocytosis of apoptotic prostate cancer cells via HIF-1α stabilization
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Project description:Peritoneal macrophages from control and Mac-Gata6 KO (LysM-cre;Gata6-floxed) mice were determined for genome wide gene expression. Sorted peritoneal macrophages from control and Mac-Gata6 KO mice were performed for whole genome expression analysis by Illumina microarray
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Project description:Clearance of apoptotic cancer cells by macrophages, known as efferocytosis, fuels the bone-metastatic growth of prostate cancer cells via pro-inflammatory and immunosuppressive mechanisms that are still unclear. In this study, single-cell transcriptomics of bone marrow macrophages undergoing efferocytosis of apoptotic prostate cancer cells revealed a significant enrichment of a cellular response to hypoxia. Here we show that efferocytic macrophages promote HIF-1α stabilization under normoxic conditions through interaction with phosphorylated STAT3. Inflammatory cytokine gene expression analysis of efferocytic HIF-1α-mutant macrophages revealed a reduced expression of the pro-tumorigenic Mif. Furthermore, stabilization of HIF-1α using the HIF-prolyl-hydroxylase inhibitor, Roxadustat, enhanced MIF expression in macrophages. Finally, macrophages treated with recombinant MIF protein activated NF-κB (p65) signaling increased the expression of pro-inflammatory cytokines. Altogether, these findings suggest that the clearance of apoptotic cancer cell by tumor-associated macrophages triggers p-STAT3/HIF-1α/MIF signaling to enhance tumor-promoting inflammation in bone, suggesting this axis as a target for metastatic prostate cancer.
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Project description:In vitro models are often used to study the functions of macrophages, including the process of phagocytosis. The use of primary macrophages has limitations associated with the individual characteristics of animals, which can lead to insufficient standardization of the results obtained. This disadvantage is lacking in immortalized cell lines. The PMJ2-R cell line was derived from in vivo immortalization and retains many of the primary peritoneal macrophages functions, but little is known about its differences from normal cells. In this article, we carried out a comparative analysis of the proteomes of PMJ2-R cells and primary peritoneal macrophages isolated from the C57BL/J6 mice. Particular attention was paid to the analysis of proteins involved in the process of phagocytosis. A total of 4005 proteins were identified, of which 797 were quantified. Our results indicate that there are significant differences in the abundance of a large number of proteins, including important proteins associated with the process of phagocytosis, such as Elmo1, Gsn, Hspa8, Itgb1, Ncf2, Rac2, Rack1, Sirpa, Sod1, C3, Msr1. Thus, when using PMJ2R cells as model cells in the study of the peritoneal macrophages functions, features revealed in this study should be taken into account.
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Project description:Purpose: Characterize the gene expression profile of wild-type and Zeb1 (+/-) peritoneal mouse macrophages through RNA sequencing Methods: RNA sequencing of RNA from peritoneal macrophages isolated by peritoneal lavage and FACS sorting (F4/80+ CD11b+) from wild-type and Zeb1 (+/-) mice (2 mice pooled for each sample/set) Results: Wild-type and Zeb1 (+/-) peritoneal mouse macrophages display differential gene expression. Zeb1 (+/-) peritoneal macrophages express lower levels of SPM (F4/80low)-associated genes Conclusions: Wild-type and Zeb1 (+/-) peritoneal mouse macrophages display differential gene expression. Zeb1 (+/-) peritoneal macrophages express lower levels of SPM (F4/80low)-associated genes
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