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MRNA expression profiles in primary microglia cells treated with R848, IFN-α, and R848+ IFN-α

ABSTRACT: Using RNA-seq, we have reported signaling crosstalk between IFN-α and R848 at the level of transcriptomes highlighting synergsitic gene expression dynamics upon different receptors signaling in microglia cells

ORGANISM(S): Mus musculus

PROVIDER: GSE104056 | GEO | 2018/09/20

REPOSITORIES: GEO

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mRNA expression profiles in primary microglia cells treated with R848, IFN-α, and R848+ IFN-α

Project description:mRNA expression profiles in primary microglia cells treated with R848, IFN-α, and R848+ IFN-α

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Microglia have a more extensive and divergent response to interferon-alpha compared with astrocytes

Project description:Type I interferons (IFN-I) are crucial for effective antimicrobial defence in the central nervous system (CNS) but also can cause severe neurological disease (termed cerebral interferonopathy) as exemplified by Aicardi-Goutières Syndrome and chronic viral infection. In the CNS, microglia and astrocytes have essential roles in host responses to infection and injury, with both cell types responding to IFN-I. However, the extent to which the IFN-I responses of these cells differ, if at all, is still unknown. Here we determined the global transcriptional responses of astrocytes and microglia to the IFN-I, IFN-alpha. MGCs were prepared from 2–4 day-old C57BL/6 mice. Purified primary astrocytes were obtained from the MGCs by magnetic activated cell sorting using anti-CD11b beads. Microglia were obtained from mixed glial cell cultures by mechanical shaking for 4 h. After treating astrocytes and microglia with IFN-alpha for 12 h, microarray using Affymetrix mouse genome array 430 2.0 array was performed on total RNA extracted from these cells. We found that under basal conditions, each cell type has a unique gene expression pattern reflective of its developmental origin and biological function. Following stimulation with IFN-alpha for 12 h, astrocytes and microglia also displayed a common core response that was characterized by the increased expression of genes required for pathogen detection and elimination. Microglia had a more extensive and diverse response to IFN-alpha with twice the number of genes upregulated (282 vs. 141 genes) when compared with astrocytes. Validation of the findings in vivo further suggested that astrocytes and microglia play important but distinct roles in the development of IFN-alpha-driven cerebral interferonopathies.

2018-03-18 | E-MTAB-6301 | biostudies-arrayexpress

Sustained R848 treatment of Mice

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2008-09-30 | GSE12957 | GEO

Transcription profiling of neonatal rat microglia after IFN-gamma activation

Project description:IFN-gamma is a classical microglial stimulant. We used microarrays to investigate the microglial gene regulatory network activated by interferon-gamma. Experiment Overall Design: Primary rat microglia cultures were established and maintained for 15 days. For activation studies, fresh media containing IFN-gamma (100 U/ml) were added and left overnight (16 hours). Total RNA was extraction and hybridized on Affymetrix microarrays (RG_U34A). Five arrays were run from total RNA derived from unstimulated 5 independent cultures (Mgl_CON1, Mgl_CON2, Mgl_CON3, Mgl_CON5 and Mgl_CON6). Three standard microglial cultures were stimulated (Mgl_IFNgamma_2 and Mgl_IFNgamma_3) with one of the samples analysed in triplicate as a technical control (Mgl_IFNgamma_1a, Mgl_IFNgamma_1b and Mgl_IFNgamma_1c).

2008-06-14 | E-GEOD-6353 | biostudies-arrayexpress

mouse BMDM poly(I:C), R848, or poly(I:C)+R848 stimulation

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2017-08-17 | GSE89987 | GEO

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The Effect of TLR2 Activation on Transcriptome of IFN-induced Microglia

Project description:In murine glioma models, TLR2 is selectively upregulated in glioma-associated microglia. We found that TLR2 activation led to down-regulation of MHC-II in microglia. Therefore, we examined the effect of TLR2 activation on IFN-gamma-induced microglial transcriptome by in vitro culture of murine adult microglia. In the study, we confirmed that TLR2 activation led to Ciita shutdown and inhibition of MHC-II related genes, which also revealed the role of TLR2 in inducing APC disability and tumor immune escape. In addition, this work can be used to reveal the transcriptional characteristics of murine microglia during IFN-gamma and TLR2 activation.

2018-09-25 | GSE109297 | GEO

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Project description:iCell human microglia were treated in triplicate (replicates a, b and c) with vehicle or 5 ng/mL human IFN-β for 6 or 24 h followed by RNA-sequencing (RNA-seq). The sample preparation, RNA-seq and transcriptomics data analysis were performed by AstraZeneca. The raw FASTQ data files cannot be made publicly available in accordance with the UK General Data Protection Regulation and the Data Protection Act 2018.

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Long noncoding RNA signatures induced by TLR7 and type I IFN signaling in activated human plasmacytoid dendritic cells

Project description:Using genome-wide sequencing approaches we viewed the contributions of Toll-like receptor 7 (TLR7) and type I interferon (IFN-I) in the regulation of coding and noncoding RNA expression in CAL-1 pDC treated with R848 or IFNβ. Functional enrichment analysis revealed both the unique and synergistic roles of TLR7 and IFN-I signaling in the orchestration of pDC function.

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Microarray analysis to evaluate the role of USP18 in primary microglia and the microglia cell line BV-2

Project description:Microglia are tissue macrophages of the central nervous system (CNS) that control tissue homeostasis, and as such they are crucially important for organ integrity. Microglia dysregulation is thought to be causal for a group of neuropsychiatric, neurodegenerative and neuroinflammatory diseases, called ‘microgliopathies’. However, how the intracellular stimulation machinery in microglia is controlled is poorly understood. By using expression studies, we identified the ubiquitin-specific protease (Usp) 18 in white matter microglia that essentially contributes to microglial quiescence under homeostatic conditions. We further found that microglial Usp18 negatively regulated the activation of STAT1 and concomitant induction of interferon-induced genes thereby disabling the termination of IFN signalling. Unexpectedly, the Usp18-mediated feedback loop was independent from the catalytic domain of the protease but instead required the interacting region of Ifnar2. Additionally, the absence of Ifnar1 completely rescued microglial activation indicating a tonic IFN signal mediated by receptor interactions under non-diseased conditions. Finally, conditional depletion of Usp18 only in myeloid cells significantly enhanced the disease burden in a mouse model of CNS autoimmunity, increased axonal and myelin damage and determined the spatial distributions of CNS lesions that shared the same STAT1 signature as myeloid cells found in active multiple sclerosis (MS) lesions. These results identify Usp18 as novel negative regulator of microglia activation, demonstrate a protective role of the IFNAR pathway for microglia and establish Usp18 as potential therapeutic target for the treatment of MS. Primary microglia (WT, USP18ko and USP18_C61A mice) and BV-2 cells (treated with control siRNA or siRNA against USP18) were incubated with 500 U/ml IFN-b. At different timepoints (0h, 6h, and 24h) RNA samples were taken and analyzed via Microarray

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