Decreased miR-4512 Levels in Monocytes and Macrophages of Individuals With Systemic Lupus Erythematosus Contribute to Innate Immune Activation and Neutrsophil NETosis by Targeting TLR4 and CXCL2.
Decreased miR-4512 Levels in Monocytes and Macrophages of Individuals With Systemic Lupus Erythematosus Contribute to Innate Immune Activation and Neutrsophil NETosis by Targeting TLR4 and CXCL2.
复制标题
系统性红斑狼疮患者单核细胞和巨噬细胞中 miR-4512 水平降低通过靶向 TLR4 和 CXCL2 促进先天免疫激活和中性粒细胞 NETosis
DOI：
10.3389/fimmu.2021.756825
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发表时间：
2021
影响因子：
7.3
通讯作者：
Deng D
中科院分区：
文献类型：
--
作者：
Yang B;Huang X;Xu S;Li L;Wu W;Dai Y;Ge MX;Yuan L;Cao W;Yang M;Wu Y;Deng D
目的系统性红斑狼疮（SLE）是一种病因复杂的自身免疫性疾病。我们的目的是阐明microRNAs（miRNAs）在藏族人群SLE中的作用机制和治疗潜力。方法采用miRNA-mRNA联合测序技术，检测SLE患者（n = 5）和健康对照者（n = 5）外周血单个核细胞中与SLE相关免疫异常相关的miRNAs。荧光素酶报告基因检测用于鉴定候选miRNA的潜在靶点。在使用多个细胞系的miRNA-agomir/Escheromir转染测定中并通过表达分析来验证靶基因。通过多种细胞因子谱评估候选miRNA对单核细胞和巨噬细胞活化的影响。通过用候选miRNA转染的单核细胞和巨噬细胞的上清液进行细胞刺激，在体外分析了神经元细胞外陷阱（NET）的形成。采用啮齿动物MRL/lpr狼疮模型评价CXCL 2Ab对系统性红斑狼疮的治疗作用以及对免疫紊乱的调节作用。结果miRNA-4512是参与调节中性粒细胞活化和趋化因子相关通路的候选miRNA。miR-4512表达在SLE患者的单核细胞和巨噬细胞中显著降低。miR-4512通过靶向TLR 4和CXCL 2抑制TLR 4通路。单核细胞和巨噬细胞miR-4512水平的降低导致多种促炎细胞因子在体外的表达。miR-4512转染的单核细胞和巨噬细胞的上清液显著促进NETs的形成（P < 0.05）。阻断CXCL 2可减轻MRL/lpr小鼠的各种致病表现，包括肾损伤和SLE免疫标志物的表达。结论miR-4512在SLE先天免疫调节中的作用。miR-4512的作用涉及通过直接靶向TLR 4和CXCL 2来调节单核细胞、巨噬细胞和NET的形成，表明miR-4512-TLR 4-CXCL 2轴是SLE中潜在的新型治疗靶点。
Objective Systemic lupus erythematosus (SLE) is an autoimmune disease with complex etiology that is not yet entirely understood. We aimed to elucidate the mechanisms and therapeutic potential of microRNAs (miRNAs) in SLE in a Tibetan population. Methods Peripheral blood mononuclear cells from SLE patients (n = 5) and healthy controls (n = 5) were used for miRNA–mRNA co-sequencing to detect miRNAs related to immune abnormalities associated with SLE. Luciferase reporter assay was used to identify potential targets of candidate miRNA. The target genes were verified in miRNA-agomir/antagomir transfection assays with multiple cells lines and by expression analysis. The effects of candidate miRNA on monocyte and macrophage activation were evaluated by multiple cytokine profiling. Neutrophil extracellular traps (NETs) formation was analyzed in vitro by cell stimulation with supernatants of monocytes and macrophages transfected with candidate miRNA. The rodent MRL/lpr lupus model was used to evaluate the therapeutic effect of CXCL2Ab on SLE and the regulation effect of immune disorders. Results Integrated miRNA and mRNA expression profiling identified miRNA-4512 as a candidate miRNA involved in the regulation of neutrophil activation and chemokine-related pathways. MiR-4512 expression was significantly reduced in monocytes and macrophages from SLE patients. MiR-4512 suppressed the TLR4 pathway by targeting TLR4 and CXCL2. Decreased monocyte and macrophage miR-4512 levels led to the expression of multiple proinflammatory cytokines in vitro. Supernatants of miR-4512 antagomir-transfected monocytes and macrophages significantly promoted NETs formation (P < 0.05). Blocking of CXCL2 alleviated various pathogenic manifestations in MRL/lpr mice, including kidney damage and expression of immunological markers of SLE. Conclusions We here demonstrated the role of miR-4512 in innate immunity regulation in SLE. The effect of miR-4512 involves the regulation of monocytes, macrophages, and NETs formation by direct targeting of TLR4 and CXCL2, indicating the miR-4512-TLR4-CXCL2 axis as a potential novel therapeutic target in SLE.