MicroRNA-204-3p inhibits lipopolysaccharide-induced cytokines in familial Mediterranean fever via the phosphoinositide 3-kinase γ pathway
MicroRNA-204-3p inhibits lipopolysaccharide-induced cytokines in familial Mediterranean fever via the phosphoinositide 3-kinase γ pathway
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DOI：
10.1093/rheumatology/kex451
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发表时间：
2018-04-01
期刊：
影响因子：
5.5
通讯作者：
Kawakami, Atsushi
中科院分区：
文献类型：
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作者：
Koga, Tomohiro;Migita, Kiyoshi;Kawakami, Atsushi
目标。我们试图鉴定FMF中的microRNA （miRNA）谱和潜在的生物标志物，并阐明它们的基因靶点，以阐明FMF的发病机制。我们使用发作期和缓解期FMF患者的血清进行了miRNA微阵列检测。然后，我们检测了toll样受体（TLR）配体刺激THP-1细胞衍生的巨噬细胞中miRNAs的表达。用脂多糖（lps）刺激THP-1细胞衍生的巨噬细胞，转染pre-miRNA，以定量炎症细胞因子的产生。为了鉴定靶基因，我们过表达了它们的miRNA，并进行了互补DNA微阵列。用报告基因构建体和前体miRNA转染，证实了对目标mrna的抑制作用。我们发现，发作时FMF患者血清中miR-204-3p明显降低。LPS刺激THP-1细胞衍生的巨噬细胞中miR-204-3p的表达被抑制，miR-204-3p的抑制显著诱导tlr4相关细胞因子的产生。生物信息学分析显示，miR-204-3p可通过调控PI3K信号通路靶向TLR通路相关基因。报告基因实验显示，miR-204-3p直接抑制PIK3CG报告基因构建体3′-UTR的荧光素酶活性。抑制PI3K γ导致FMF患者单核细胞中IL-6和IL-12p40的含量降低。这些数据表明，血清miR-204-3p有潜力作为FMF患者有用的生物标志物，并且miR-204-3p通过靶向PI3K γ途径抑制FMF中炎症细胞因子的产生。
Objective. We sought to identify the microRNA (miRNA) profile and potential biomarkers in FMF and to clarify their gene targets to elucidate the pathogenesis of FMF.Methods. We performed an miRNA microarray using serum from FMF patients in attack and in remission. We then examined the expression of miRNAs in macrophages derived from THP-1 cells stimulated with toll-like receptor (TLR) ligands. Macrophages derived from THP-1 cells transfected with pre-miRNA were stimulated with lipopolysaccharides (LPSs) for the quantification of inflammatory cytokine production. To identify the target genes, we overexpressed their miRNA and performed a complementary DNA microarray. Transfection with reporter construct and the precursor miRNA was performed to confirm the suppression of target mRNA.Results. We found that miR-204-3p was greatly decreased in the serum from FMF patients in attack. The expression of miR-204-3p was suppressed by LPS stimulation in the macrophages derived from THP-1 cells and the inhibition of miR-204-3p significantly induced the production of TLR4-related cytokines. The bioinformatic analysis showed that miR-204-3p is predicted to target genes implicated in the TLR pathway through the regulation of PI3K gamma signalling. The reporter assay revealed that miR-204-3p directly suppressed the luciferase activity of 3'-UTR of PIK3CG reporter construct. The inhibition of PI3K gamma resulted in decreased amounts of IL-6 and IL-12p40 in monocytes from FMF patients.Conclusion. These data suggest that serum miR-204-3p has potential as a useful biomarker in FMF patients and that miR-204-3p serves as a suppressor of inflammatory cytokine production in FMF by targeting the PI3K gamma pathway.