An interferon-like small chemical compound CDM-3008 suppresses hepatitis B virus through induction of interferon-stimulated genes
An interferon-like small chemical compound CDM-3008 suppresses hepatitis B virus through induction of interferon-stimulated genes
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DOI：
10.1371/journal.pone.0216139
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发表时间：
2019-06-12
期刊：
影响因子：
3.7
通讯作者：
Kojima, Soichi
中科院分区：
文献类型：
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作者：
Furutani, Yutaka;Toguchi, Mariko;Kojima, Soichi
口服核苷酸类似物和注射干扰素-α（IFN α）用于实现对B型肝炎病毒（HBV）复制的立即抑制。核苷酸类似物和IFN α分别通过诱导APOBEC 3脱氨酶家族分子去除共价闭合环状DNA（cccDNA），抑制病毒聚合酶活性并导致病毒的长期根除。本研究旨在探索可口服的低分子量药物CDM-3008（RO 8191）（通过结合IFN α/β受体2（IFNAR 2）和激活JAK/STAT途径模拟IFN α）是否可以抑制HBV复制并降低cccDNA水平。在原代培养的人肝细胞中，CDM-3008处理后HBV DNA水平以剂量依赖性方式降低，半数最大抑制浓度（IC 50）值为0.1 μ M，同时伴随细胞cccDNA水平、细胞培养基中HBeAg和HBsAg水平的显著降低。使用微阵列，我们全面分析和比较CDM-3008和IFN α处理的原代培养人肝细胞中基因（mRNA）表达的变化。如前所述，CDM-3008模拟参与干扰素信号传导途径的基因的诱导。OAS 1和ISG 20 mRNA表达通过CDM-3008和IFN α类似地增强。因此，CDM-3008可抑制pgRNA表达以显示抗HBV活性。CDM-3008和IFN α处理也诱导了APOBEC 3F和3G mRNA表达，表明cccDNA可以通过诱导的APOBEC 3家族蛋白降解。我们鉴定了与IFN α处理的细胞相比，CDM-3008处理的细胞中表达特异性增强的基因。抑制STAT活化的SOCS 1、SOCS 2、SOCS 3和CISH的表达在CDM-3008处理的细胞中增强，表明与IFN α处理的细胞相比，在CDM-3008处理的细胞中JAK/STAT途径的反馈抑制增强。此外，CDM-3008与临床使用的核苷恩替卡韦在抑制HBV复制方面显示出累加效应。总之，CDM-3008通过激活JAK/STAT途径显示出抗HBV活性，诱导干扰素刺激基因（ISG）的表达，具有比IFN α更大的反馈抑制作用。
Oral administration of nucleotide analogues and injection of interferon-alpha (IFN alpha) are used to achieve immediate suppression in replication of hepatitis B virus (HBV). Nucleotide analogs and IFN alpha inhibit viral polymerase activity and cause long-term eradication of the virus at least in part through removing covalently closed circular DNA (cccDNA) via induction of the APOBEC3 deaminases family of molecules, respectively. This study aimed to explore whether the orally administrable low molecular weight agent CDM-3008 (RO8191), which mimics IFN alpha through the binding to IFN alpha/beta receptor 2 (IFNAR2) and the activation of the JAK/STAT pathway, can suppress HBV replication and reduce cccDNA levels. In primary cultured human hepatocytes, HBV DNA levels were decreased after CDM-3008-treatment in a dose-dependent manner with a half-maximal inhibitory concentration (IC50) value of 0.1 mu M, and this was accompanied by significant reductions in cellular cccDNA levels, both HBeAg and HBsAg levels in the cell culture medium. Using a microarray we comprehensively analyzed and compared changes in gene (mRNA) expression in CDM-3008- and IFN alpha-treated primary cultured human hepatocytes. As reported previously, CDM-3008 mimicked the induction of genes that participate in the interferon signaling pathway. OAS1 and ISG20 mRNA expression was similarly enhanced by both CDM-3008 and IFN alpha. Thus, CDM-3008 could suppress pgRNA expression to show anti-HBV activity. APOBEC3F and 3G mRNA expression was also induced by CDM-3008 and IFN alpha treatments, suggesting that cccDNA could be degraded through induced APOBEC3 family proteins. We identified the genes whose expression was specifically enhanced in CDM-3008-treated cells compared to IFN alpha-treated cells. The expression of SOCS1, SOCS2, SOCS3, and CISH, which inhibit STAT activation, was enhanced in CDM-3008-treated cells suggesting that a feedback inhibition of the JAK/STAT pathway was enhanced in CDM-3008-treated cells compared to IFN alpha-treated cells. In addition, CDM-3008 showed an additive effect with a clinically-used nucleoside entecavir on inhibition of HBV replication. In summary, CDM-3008 showed anti-HBV activity through activation of the JAK/STAT pathway, inducing the expression of interferon-stimulated genes (ISGs), with greater feedback inhibition than IFN alpha.