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Homo sapiens

ABSTRACT: RNA sequencing of macrophages derived from monocytes differentiated in presence of M-CSF, GM-CSF and heat killed mycobacteria M. obuense

PROVIDER: PRJNA397874 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15050902.fastq.gz	Fastqsanger.gz
	SRR15050903.fastq.gz	Fastqsanger.gz
	SRR15050904.fastq.gz	Fastqsanger.gz
	SRR15050905.fastq.gz	Fastqsanger.gz
	SRR15050906.fastq.gz	Fastqsanger.gz

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RNA sequencing of macrophages derived from monocytes differentiated in presence of M-CSF, GM-CSF and heat killed mycobacteria M. obuense

Project description:Earlier work has shown that macrophages derived from differentiation of monocytes by heat killed (HK) mycobacteria (e.g. M. obuense) exhibit unique immunophenotypic and molecular properties. Yet, our knowledge of these properties is still limited. The goal of the study is to understand global gene expression programs that are differentially modulated in macrophages derived from monocytes that were differentiated through three different routes: in presence of M-CSF, GM-CSF and the heat killed mycobacterium M. obuense. We performed RNA sequencing (RNA-Seq) of monocyte-derived macrophages (MDM) that were acquired from four separate healthy donors and differentiated by incubation with M-CSF (M-MDM), GM-CSF (GM-MDM) and HK M. obuense (Mob-MDM) (n = 12 samples). We report that Mob-MDM exhibit unique gene expression programs that may explain its unique immunophenotypic properties and thus immunomodulatory capacity.

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Project description:Human monocyte-derived macrophages (MDM) from HDM-allergic donors were compared to MDM from healthy donors. CD14+ monocytes were isolated from donated blood and differentiated in the presence of GM-CSF and TGFb (alveolar-like MDM) for 7 days before havest of RNA

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Methylation data and effect of JAK3 inhibition on human MO-to-DC and MO-to-MAC differentiation

Project description:Comparison of the DNA methylation profiles of CD14+ monocytes from human peripheral blood with derived dendritic cells (DCs) and macrophages (MACs) obtained by exposure with GM-CSF/IL-4 and GM-CSF, respectively. Effects on the methylation profiles of DCs and MACs of JAK3 inhibitor PF-956980 The methylation profiles of bisulfite-modified DNA of human CD14+ monocytes were compared with derived dendritic cells (DCs), macrophages (MACs) following GM-CSF/IL-4 and GM-CSF incubation, and DC and MAC samples incubated with JAK3 inhibitor PF-956980 using the Infinium HumanMethylation450 BeadChips (Illumina, Inc., San Diego, CA,). This platform allows the interrogation of >485,000 methylation sites per sample at single-nucleotide resolution, and comprises an average of 17 CpG sites per gene in the 99% of RefSeq genes. 96% of CpG islands are covered, with additional coverage in CpG island shores and the regions flanking them. The samples were hybridized in the array following the manufacturerÃ¢ÂÂs instructions. Total DNA isolated by standard procedures from CD14+ cells (total monocytes, MOs) corresponding to three sets of samples of monocytes (MOs), derived DCs and MACs (DCs and iMACs; DMSO as these samples were differentiated in the absence of JAK3 inhibitors) and DCs and MACs differentiated in the presence of JAK3 inhibitor PF-956980.

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Danger signal initiates differentiation of monocytes into osteopontin‑producing macrophages III

Project description:Purpose: We report the application of single-cell RNA sequencing for profiling the genexpression of different monocyte types in order to characterize monocytes that differentiate into macrophages under danger signal in comparison to conventional GM-CSF and M-CSF macrophages Methods: Monocytes freshly isolated from PBMCs were differentiated into GM-CSF, M-CSF, and danger-signal induced macrophages. Two million cells of each macrophage population were tagged accordingly with TotalSeq™‑B Hashtags. Single cell RNA sequencing was performed on all macrophage populations in order to characterize cells from each population. ScRNAseq analysis was performed with the R package Seurat on 16470 barcodes after filtering. Results: Cluster analysis reveals distinct separation of all three macrophage populations. Conclusion: The clear difference in overall gene expression profile suggests that macrophages exposed to danger signal differentiate into a novel macrophage type in contrast to conventional GM-CSF and M-CSF monocytes.

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Project description:Caspases, which are key effectors of apoptosis, have demonstrated non-apoptotic functions. One of these functions is the differentiation into macrophages of peripheral blood monocytes exposed to Colony-Stimulating Factor-1 (CSF1). Conversely, GM-CSF induces the differentiation of monocytes into macrophages in a caspase-independent manner. Macrophages generated by CSF1 and GM-CSF have distinct polarity. Macrophage polarization plays an important role in the pathogenesis of diverse human diseases as cancer, leading us to explore if caspase inhibition would affect macrophage polarization. To explore the role of caspases in CSF1 differentiation, we used human monocytes sorted from buffy coats treated by cytokines. We reported that caspase inhibition delays the ex vivo differentiation of peripheral blood monocytes exposed to CSF1 and modifies the phenotype of generated macrophages, e.g. cell shape, surface markers. Moreover, by RNAseq, we observed that the macrophages generated in presence of CSF1 and QVD are different from CSF1-treated monocytes and from GM-CSF-treated monocytes. Cell cycle and focal adhesion-related pathway genes were selectively down-regulated. This study confirms the importance of caspase activation in CSF1 differentiation.

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