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Characterization of neutrophil extracellular trap-associated RNA, a novel DAMP acting in concert with the antimicrobial peptide LL37 as intentional but self-limiting drivers of inflammation

ABSTRACT: Neutrophil extracellular traps (NETs) are a key antimicrobial feature of cellular innate immunity mediated by polymorphonuclear neutrophils (PMNs). NETs trap and kill microbes but have also been linked to inflammation in atherosclerosis, arthritis, or psoriasis by unknown mechanisms. We here report that NET-associated RNA (naRNA, which was RNA-sequenced from human primary neutrophil NETs here) stimulated further NET formation in naïve PMNs via a unique TLR8-NLRP3-caspase-1-gasdermin D-dependent inflammasome pathway. Keratinocytes also responded to naRNA with expression of psoriasis-related genes (e.g. IL17, IL36) via atypical NOD2-RIPK signaling. In vivo naRNA drove skin inflammation, which was drastically ameliorated by genetic ablation of RNA sensing. The naRNA-LL37 ‘composite DAMP’ was pre-stored in resting neutrophil granules, defining sterile NETs as intentionally inflammatory webs that amplify neutrophil activation. However, the activity of the naRNA-LL37 DAMP was transient and hence supposedly self-limiting under physiological conditions. Only upon dysregulated NET release like psoriasis, TLR-NLRP3-mediated naRNA sensing may represent both potential cause of disease and new intervention target.

ORGANISM(S): Homo sapiens

PROVIDER: GSE253440 | GEO | 2024/05/06

REPOSITORIES: GEO

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Human lupus netting neutrophils induce endothelial damage, infiltrate tissues and expose immunostimulatory molecules

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2011-06-22 | E-GEOD-26975 | biostudies-arrayexpress

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2023-11-23 | PXD025702 | Pride

Human lupus netting neutrophils induce endothelial damage, infiltrate tissues and expose immunostimulatory molecules

Project description:Our group has proposed that low-density granulocytes (LDGs) play an important role in lupus pathogenesis, as they can damage endothelial cells and synthesize increased levels of proinflammatory cytokines and type I interferons. LDGs have a heightened capacity to synthesize neutrophil extracellular traps (NETs). NETs from LDGs display increased levels of bactericidal and immunostimulatory proteins, such as the cathelicidin LL37 and externalize double-stranded DNA (dsDNA). Lupus netting LDGs have increased capacity to kill endothelial cells and expose IL-17. Through NETosis, lupus neutrophils stimulate plasmacytoid DCs to synthesize IFN-α. Our results further expand the potential pathogenic role of aberrant lupus neutrophils through a NET-mediated effect. We used microarrays to analyze the gene expression of neutrophils in healthy and lupus patients, and of low-density granulocytes in lupus patients.

2011-06-22 | GSE26975 | GEO

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NETOME: The Molecular Characterization of Neutrophil Extracellular Traps (NETs)

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Characterization of Early-Phase Neutrophil Extracellular Traps in Urinary Tract Infections

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Proteomic analysis of neutrophil extracellular traps (healthy control, rheumatoid arthritis, systemic lupus erythematosus)_C4PR_LIV

Project description:Neutrophil Extracellular Traps (NETs) are chromatin-derived extracellular structures that are expelled from neutrophils in response to infectious or inflammatory stimuli. NET DNA structures are decorated with proteins including histones, myeloperoxidase and neutrophil elastase. NETs are implicated in the development of auto-immunity in diseases including rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) through the externalisation of intracellular neoepitopes e.g. dsDNA and nuclear proteins in SLE and citrullinated peptides in RA. The aim of this work was to use quantitative proteomics to identify and measure NET proteins produced by neutrophils from healthy individuals, and from patients with RA and SLE.

2019-02-18 | PXD011796 | Pride

NET induced by different stimuli: a comparative proteomic analysis

Project description:Neutrophil extracellular traps (NET) formation is part of the neutrophil response to infections, but excessive or inappropriate NETosis may trigger the production of autoantibodies and cause organ damage in autoimmune disorders. Spontaneously netting neutrophils are not frequent and induction of NET in vitro by selected stimuli is necessary to investigate their structure. In the present work, the protein composition and post-translational modifications of NET produced under different stimuli is studied by means of proteomic analysis. Neutrophils from healthy donors were stimulated by PMA, A23187, E.Coli LPS or untreated; after 3 hours cells were washed, treated with DNase and supernatants collected for mass spectrometry. Data were analyzed by unsupervised hierarchical clustering analyses. We identified proteins contained in NETs of any source or exclusive of one stimulus: LPS-induced and spontaneous NET diverge in protein composition, while PMA- and A23187-induced NET appear more similar. Among the post-translational modifications we examined, methionine sulfoxidation is frequent especially in PMA- and LPS-induced NETs. Myeloperoxidase is the protein more extensively modified. Thus, proteomic analysis indicates that NETs induced by different stimuli are heterogeneous in terms of both protein composition and post-translational modifications, suggesting that NET induced in different conditions may have different biological effects.

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