Project description:miRNA expression profiling of human monocyte-derived dendritic cells (moDCs) during maturation. Immature, 4h and 16h LPS-activated moDCs were used.
Project description:mouse Freshly isolated monocyte (LY6C high) and (LY6C low) were purified from WT DBA/1 mice blood, and sorted by FACS. We used Taqman miRNA TLDA arrays to performed miRNA profiling
Project description:MicroRNAs (miRNAs, miRs) modulate a multitude of cellular events. Here, we identify functional miRNA-protein networks that regulate human monocyte-derived dendritic cell (MDDC) differentiation. MiRNA profiling revealed stage-specific differential expression of 20 miRNAs during days 1, 3 and 5 of MDDC differentiation. To identify and prioritize miRNA-protein networks for functional validation, we developed a target ranking algorithm that incorporates many features of miRNA regulatory networks. This system prioritized miR-21, miR-34a, and their cognate targets WNT1 and JAG1 for functional validation. Inhibition of both miR-21 and miR-34a stalled MDDC differentiation, as quantified by DC-SIGN/CD14 expression ratios, showing cooperative involvement of these miRNAs in MDDC differentiation. We confirmed that the 3’ UTRs of WNT1 and JAG1 were functional targets of these miRNAs and provide evidence that these targets were translationally suppressed. Significantly, exogenously added Wnt-1 and Jagged-1 also stalled MDDC differentiation, suggesting that miRNA mediated inhibition of endogenous WNT1 and JAG1 expression was important for proper MDDC differentiation. Finally, inhibition of miR-21 and miR-34a, or addition of Wnt-1 and Jagged-1 led to a decrease in endocytic capacity, a key function of immature DCs. Thus, our novel approach identified and validated some miRNA-protein networks involved in phenotypic and functional MDDC differentiation.
Project description:Human Freshly isolated monocyte (CD14++CD16-) and (CD14+ CD16++) were purified from healthy volunteers' blood, and sorted by FAC. We used Taqman miRNA TLDA arrays to performed miRNA profiling
Project description:To elucidate the classification and function of monocyte subsets in the neuroprotective retina, we used single-cell RNA sequencing on isolated retinal immune cells. We identified a novel subpopulation of monocyte named Ly6ClowMo/MΦ, which display neural restorative features.
Project description:Purpose: Identify differences in gene expression profiles in fetal monocytes - cells that persist and differentiate postnatally - according to distinct placental histologic domains. Methods: We first isolated classical and intermediate monocyte subsets via FACS and performed transcriptomic profiling of 140 samples (70 classical and 70 intermediate monocyte samples) using bulk RNA-Seq. Results: We report that placental lesions are associated with gene expression changes in fetal monocyte subsets. Specifically,fetal monocytes exposed to acute placental inflammation upregulate biological processes related to monocyte activation, monocyte chemotaxis, and platelet function while monocytes exposed to maternal vascular malperfusion lesions downregulate these processes. Additionally, we show that intermediate monocytes might be a source of mitogens, such as HBEGF, NRG1, and VEGFA, implicated in different outcomes related to prematurity. Conclusions: This is the first study to show that placental lesions are associated with unique changes in fetal monocytes and monocyte subsets. As fetal monocytes persist and differentiate into various phagocytic cells following birth, our study may provide insight into morbidity related to prematurity and ultimately potential therapeutic targets.
Project description:miRNA expression profiling of human monocyte-derived dendritic cells (moDCs) during maturation. Immature, 4h and 16h LPS-activated moDCs were used. In this study were analysed 2 biological samples of RNA extracted from 4h-post LPS stimulation moDCs and 2 bfrom 16h-post LPS stimulation. The RNA from 0h-post LPS stimulation were used as reference sample. Were also performed dye-swaps of each biological sample as technical replicates.