MiR-155 Regulates PAD4-Dependent Formation of Neutrophil Extracellular Traps
MiR-155 Regulates PAD4-Dependent Formation of Neutrophil Extracellular Traps
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DOI：
10.3389/fimmu.2019.02462
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发表时间：
2019-11-01
影响因子：
7.3
通讯作者：
Thorlacius, Henrik
中科院分区：
文献类型：
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作者：
Hawez, Avin;Al-Haidari, Amr;Thorlacius, Henrik
越来越多的数据表明，中性粒细胞胞外陷阱（NET）在多种疾病中发挥着关键作用。肽基精氨酸脱亚胺酶 4 (PAD4) 通过组蛋白瓜氨酸化调节 NET 形成。本研究的目的是检查 miR-155 在控制 PAD4 依赖性 NET 生成中的作用。用 PMA 和 MIP-2 刺激骨髓中性粒细胞。中性粒细胞与翻译抑制剂（放线菌酮或嘌呤霉素）预孵育可显着减少 PMA 或 MIP-2 诱导的 NET 形成。用模拟 miR-155 转染中性粒细胞可增加 PMA 诱导的 PAD4 mRNA 表达和 NET 形成。相比之下，用 antagomiR-155 转染可减少 PAD4 mRNA 和 NET 响应 PMA 攻击的诱导。 PAD4 的生物信息学检查揭示了 3'-UTR 区域富含 AU 的元件的潜在结合位点。通过使用靶位点阻断剂和 RNA 免疫沉淀检查了 miR-155 与 PAD4 的结合，揭示了 miR-155 对 PAD4 mRNA 的调节是通过 3'-UTR 区域中富含 AU 的元件介导的。总之，我们的研究结果表明 miR-155 正向调节中性粒细胞 PAD4 的表达和细胞外陷阱的排出。因此，我们的新结果表明，靶向 miR-155 可能有助于抑制炎症性疾病中过度的 NET 生成。
Accumulating data suggest that neutrophil extracellular traps (NETs) exert a key function in several diseases. Peptidylarginine deiminase 4 (PAD4) regulates NET formation via citrullination of histones. The aim of this study was to examine the role of miR-155 in controlling PAD4-dependent generation of NETs. Bone marrow neutrophils were stimulated with PMA and MIP-2. Pre-incubation of neutrophils with translational inhibitors (cycloheximide or puromycin) markedly decreased NET formation induced by PMA or MIP-2. Neutrophil transfection with a mimic miR-155 increased PMA-induced PAD4 mRNA expression and NET formation. In contrast, transfection with an antagomiR-155 decreased induction of PAD4 mRNA and NETs in response to PMA challenge. Bioinformatical examination of PAD4 revealed a potential binding site in AU-rich elements at the 3 '-UTR region. MiR-155 binding to PAD4 was examined by use of target site blockers and RNA immunoprecipitation, revealing that miR-155 regulation of PAD4 mRNA is mediated via AU-rich elements in the 3 '-UTR region. In conclusion, our findings demonstrate that miR-155 positively regulates neutrophil expression of PAD4 and expulsion of extracellular traps. Thus, our novel results indicate that targeting miR-155 might be useful to inhibit exaggerated NET generation in inflammatory diseases.