IFN-λ Inhibits MiR-122 Transcription through a Stat3-HNF4α Inflammatory Feedback Loop in an IFN-α Resistant HCV Cell Culture System.
IFN-λ Inhibits MiR-122 Transcription through a Stat3-HNF4α Inflammatory Feedback Loop in an IFN-α Resistant HCV Cell Culture System.
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DOI：
10.1371/journal.pone.0141655
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Dash S
中科院分区：
文献类型：
--
作者：
Aboulnasr F;Hazari S;Nayak S;Chandra PK;Panigrahi R;Ferraris P;Chava S;Kurt R;Song K;Dash A;Balart LA;Garry RF;Wu T;Dash S
在持续感染的细胞培养中，丙型肝炎病毒复制仍然对干扰素-α/RBV联合治疗具有抵抗力，而干扰素-λ1诱导病毒清除。干扰素-λ1诱导丙型肝炎病毒持续清除的抗病毒机制尚未确定。目的探讨干扰素-λ在丙型肝炎病毒细胞培养模型中清除丙型肝炎病毒复制的机制。用等浓度的干扰素-α或干扰素-α处理敏感细胞(S3-GFP)和耐药细胞(R4-GFP)。通过检测丙型肝炎病毒复制、流式细胞仪检测丙型肝炎病毒绿色荧光蛋白表达和实时定量RT-α检测病毒λ水平，比较干扰素-GFP和干扰素-GFP 1的相对抗病毒作用。检测S3-GFP和R4-GFP细胞中Jak-Stat信号的激活、干扰素刺激基因(ISG)的表达和miRNA-122的转录。我们发现，干扰素-λ-1通过Jak-Stat信号通路以剂量依赖的方式诱导干扰素-α耐药和敏感复制子细胞清除丙型肝炎病毒，并诱导STAT 1和STAT 2的激活、ISRE-荧光素酶启动子的激活和ISG的表达。STAT3的激活也参与了干扰素-λ-1诱导的丙型肝炎病毒细胞的抗病毒活性。干扰素-λ1通过miR-24磷酸化降低R4-GFP细胞中肝细胞核因子4α(HNF4α)的表达。HNF4miR-α表达降低与miR-122表达降低有关，从而产生抗丙型肝炎病毒的作用。Northern印迹分析证实，干扰素-绿色荧光蛋白1降低了R4-λ细胞中miR-122的水平。我们的结果表明，在丙型肝炎病毒细胞培养中，干扰素-λ1激活了Stat3-HNF4miR122的α反馈炎症环，从而抑制了miR122的转录。除了经典的JAK-STAT抗病毒信号通路外，干扰素-λ1还通过炎症STAT3-HNF4α反馈环抑制miRNA122的转录，从而抑制丙型肝炎病毒的复制。在慢性炎症期间，由IFN激活的炎症反馈电路使无反应者暴露于肝细胞癌的风险中。
HCV replication in persistently infected cell culture remains resistant to IFN-α/RBV combination treatment, whereas IFN-λ1 induces viral clearance. The antiviral mechanisms by which IFN-λ1 induces sustained HCV clearance have not been determined. To investigate the mechanisms by which IFN-λ clears HCV replication in an HCV cell culture model. IFN-α sensitive (S3-GFP) and resistant (R4-GFP) cells were treated with equivalent concentrations of either IFN-α or IFN-λ. The relative antiviral effects of IFN-α and IFN-λ1 were compared by measuring the HCV replication, quantification of HCV-GFP expression by flow cytometry, and viral RNA levels by real time RT-PCR. Activation of Jak-Stat signaling, interferon stimulated gene (ISG) expression, and miRNA-122 transcription in S3-GFP and R4-GFP cells were examined. We have shown that IFN-λ1 induces HCV clearance in IFN-α resistant and sensitive replicon cell lines in a dose dependent manner through Jak-Stat signaling, and induces STAT 1 and STAT 2 activation, ISRE-luciferase promoter activation and ISG expression. Stat 3 activation is also involved in IFN-λ1 induced antiviral activity in HCV cell culture. IFN-λ1 induced Stat 3 phosphorylation reduces the expression of hepatocyte nuclear factor 4 alpha (HNF4α) through miR-24 in R4-GFP cells. Reduced expression of HNF4α is associated with decreased expression of miR-122 resulting in an anti-HCV effect. Northern blot analysis confirms that IFN-λ1 reduces miR-122 levels in R4-GFP cells. Our results indicate that IFN-λ1 activates the Stat 3-HNF4α feedback inflammatory loop to inhibit miR-122 transcription in HCV cell culture. In addition to the classical Jak–Stat antiviral signaling pathway, IFN-λ1 inhibits HCV replication through the suppression of miRNA-122 transcription via an inflammatory Stat 3–HNF4α feedback loop. Inflammatory feedback circuits activated by IFNs during chronic inflammation expose non-responders to the risk of hepatocellular carcinoma.