bioRxiv Science⌕ Search

Biology subjects

Wingert, C.

Publications and source records attributed to Wingert, C..

3 recordsLinked to original sources

Sexual dimorphism in zymosan-induced arthritis is linked with a higher IFN response in myeloid cell subsets that faithfully recapitulate RA synovial cell clusters

A key feature of rheumatoid arthritis (RA) is sexual dimorphism, with a higher incidence of RA in females. Myeloid cells are key drivers of the inflammatory effector phase of RA, but little is known about their contribution to a sexually dimorphic arthritis phenotype. We wished to utilize an arthritis model that recapitulates RA pathogenic myeloid cell subsets defined by single cell transcriptome analysis to investigate sex differences in human disease-relevant cell types and related molecular pathways. We developed a computational strategy that rigorously maps scRNAseq-defined mouse arthritis myeloid cells and clusters onto previously defined RA subsets. Synovial myeloid cells in the zymosan-induced arthritis (ZIA) model closely recapitulated four pathogenic RA myeloid cell subsets, strongly supporting the disease-relevance of the ZIA model. ZIA also effectively modeled myeloid cells in immune checkpoint inhibitor arthritis (ICI-A). These RA, ICI-A and ZIA myeloid cells express genes in TNF-NF-kB, PGE2, and IFN-STAT pathways. In ZIA, an induction phase dominated by cell clusters expressing NF-kB and PGE2 pathways transitioned to peak arthritis characterized by cell subsets co-expressing NF-kB and IFN pathways, as occurs in RA and various inflammatory diseases. Female mice exhibited increased arthritis linked with early induction of interferon-stimulated genes and increased expansion of IFN signature-expressing cell subsets. Our study identifies a computational strategy and animal model that enables investigation of recently described pathogenic myeloid cell subsets, and links increased arthritis in female mice with specific myeloid cell subsets that exhibit a stronger IFN response.

immunology↗

Epigenetic Regulation of Inflammatory NF-κB Target Genes by IFN-γ via IRF1

The regulation of inflammatory gene expression involves complex interactions between transcription factors (TFs), signaling pathways and epigenetic chromatin-mediated mechanisms. This study investigated mechanisms by which by IFN-{gamma}-mediated priming augments TLR-induced expression of NF-{kappa}B target genes in primary human monocytes. IFN-{gamma} priming enhanced the expression of signature inflammatory genes such as IL6, TNF, IL1B, and CXCL10 when monocytes were exposed to various TLR agonists. RNA-seq analysis identified genes synergistically activated by IFN-{gamma} and LPS, which were enriched in inflammatory pathways. Similar synergistic activation was observed with the TLR1/2 agonist PAM3CYS, suggesting a shared regulatory mechanism. ATAC-seq analysis revealed that TLR ligands induce IRF1 TF activity independently of IFN-{gamma}. JAK1/2 inhibitor (iJAK) treatment reduced IRF1 expression and protein levels, especially in IFN-{gamma}-treated monocytes, but not in LPS-stimulated monocytes, suggesting LPS-induced IRF1 may compensate for loss of IFN-{gamma}-induced IRF1. We applied CRISPR-Cas9 to knock out IRF1 in primary human monocytes and found loss of IRF1 abrogates synergistic activation of key inflammatory genes, suggesting a pivotal role for IRF1. This genetic data was corroborated by IRF1 CUT&RUN data showing resistance of IRF1 binding to JAK inhibition under (IFN-{gamma} + LPS) costimulated conditions, and co-occupancy of IRF1 binding sites by NF-{kappa}B. This study enhances our understanding of inflammatory gene regulation, highlighting IRF1 as a key player and a potential therapeutic target for inflammatory diseases.

immunology↗

IL-10 Targets IRFs to Suppress IFN and Inflammatory Response Genes by Epigenetic Mechanisms

Interleukin-10 (IL-10) is pivotal in suppressing inflammation and innate immune activation, in large part by suppressing induction of genes by potent inflammatory factors such as TLR ligands. Despite decades of research, molecular mechanisms underlying this inhibition have not been resolved. This study utilized an integrated epigenomic analysis of gene transcription, chromatin accessibility, histone modifications and transcription factor binding to investigate IL-10-mediated suppression of LPS and TNF responses in primary human monocytes. Instead of inhibiting core TLR4-activated pathways such as NF-{kappa}B, MAPK-AP-1 and TBK1-IRF3 signaling, IL-10 targeted IRF transcription factor activity and DNA binding, particularly IRF5 and an IRF1-mediated amplification loop that is operative in monocytes. This resulted in suppression of inflammatory NF-{kappa}B target genes, in whose activation IRFs play an amplifying role, and near-complete suppression of interferon-stimulated genes. Mechanisms of TLR4 and TNFR target gene inhibition included downregulation of chromatin accessibility, de novo enhancer formation, and IRF1-associated H3K27ac activating histone marks. These results provide a mechanism by which IL-10 suppresses inflammatory NF-{kappa}B target genes, highlight the role of IRF1 in inflammatory gene expression, and describe an underappreciated suppression of IFN responses by epigenetic mechanisms.

immunology↗