MicroRNA-223 inhibits neutrophil extracellular traps formation through regulating calcium influx and small extracellular vesicles transmission

Abstract

Modulation of miRNAs and neutrophil extracellular traps (NETs) formation are both implicated in inflammatory disorders. Adult-onset Still’s disease (AOSD) is a systemic autoinflammatory disease with neutrophilic leukocytosis and unknown etiology. Although the NETs formation is elevated in AOSD patients, the regulatory roles of miRNAs in NETs formation in AOSD remains unclear. We revealed that the circulating levels of IL-18, NETs, and miR-223 were significantly higher in active AOSD patients, compared with inactive AOSD patients or healthy controls (P < 0.005). Moreover, IL-18 increased calcium influx into neutrophils, which led to mitochondrial ROS (mROS) production and NETs formation. Elevated levels of NETs-DNA could induce miR-223 expression in neutrophils through activating Toll-like receptor 9. The upregulated miR-223 expression in neutrophils suppressed mROS production by blocking calcium influx, and subsequently inhibited IL-18-mediated NETs formation. Besides, the increased neutrophil-derived exosomal miR-223 levels were observed in active AOSD patients compared with healthy controls (P < 0.005). Our in vitro assays demonstrated that the neutrophil-derived small extracellular vesicles carried miR-223, which could repress IL-18 production in macrophages. Together, these results suggest a fine-tuned mechanism between inflammatory (IL-18 induced NETs) and anti-inflammatory (miR-223) factors in AOSD. MiR-223, mROS inhibitors, and calcium channel blockers are the potential therapeutics for autoinflammatory diseases such as AOSD.

An Update on the Pathogenic Role of Neutrophils in Systemic Juvenile Idiopathic Arthritis and Adult-Onset Still’s Disease

“…Another molecule involved in this process is microRNA–223, which modulates the immune response of the host. In this study, a fine-tuned mechanism was proposed in which IL–18, microRNA–223, and NET formation regulate inflammation in AOSD, in which NET formation inhibits calcium influx into neutrophils by upregulation of microRNA–223, and it eventually inhibits IL–18 mediated NET formation [ 110 ]. The role of LDGs and NET formation in AOSD was also identified, and it has been reported that the concentration of high mobility group box–1 protein (HMGB–1) and cathelicidin LL–37 in NETs was increased in patients with AOSD [ 111 ].…”

Bibliometrics analysis on the research status and trends of adult-onset Still’s disease: 1921-2021

“…Serum CXCL10, CXCL13 ( 54 ) and S100A12 ( 55 ) levels can be used as clinical markers to assess the disease activity of AOSD. Chen, D.Y’s team found that there is a fine-tuned mechanism between inflammatory and anti-inflammatory factors in AOSD ( 56 ), and IL-18 is an important predictor of active AOSD ( 57 ). In addition, galectin-3 ( 58 ), C-Type Lectin Domain Family 5-Member A ( 59 ), B19-NS1 ( 60 ), microRNA-134 ( 61 ), and the NLRP3 inflammasome ( 62 ) are involved in the pathogenesis of AOSD.…”

Adult-onset Still’s disease: A disease at the crossroad of innate immunity and autoimmunity

“…S100 A8/A9 and A12 are parts of NETs, function as DAMPs, and act as the ligand of TLR4 or RAGE, involved in the pathogenesis of AOSD by enhancing the feedback loop (36,48,49). IL-18 is widely reported to be increased in the serum of AOSD patients, and an updated study found that it regulates mitochondrial ROS generation by increasing calcium influx to induce the formation of NETs in a process which could be suppressed by microRNA-223 (50). Hu and colleagues reported a gene of leukocyte immunoglobulin-like receptor (LIR)-A3 (LILRA3) that plays a pathogenic role in AOSD and speculated that the functional LILRA3 is a new genetic susceptibility factor for neutrophil activation and NETosis in AOSD (51) conducted a study that revealed increased type I interferon (IFN) could induce NETs, especially ones containing oxidized mitochondrial DNA in AOSD (46).…”