Gailhouste Luc, Sudoh Masayuki, Qin Xian-Yang, Watashi Koichi, Wakita Takaji, Ochiya Takahiro, Matsuura Tomokazu, Kojima Soichi, Furutani Yutaka
Liver Cancer Prevention Research Unit, RIKEN Cluster for Pioneering Research, Wako, Japan.
Department of Translational Research, Joint Research Center for Human Retrovirus Infection, Kagoshima University, Kagoshima, Japan.
Cell Death Discov. 2021 Jun 2;7(1):130. doi: 10.1038/s41420-021-00515-y.
Chronic hepatitis B virus (HBV) infections remain a health burden affecting ~250 million people worldwide. Thus far, available interferon-alpha (IFNα)-based therapies have shown unsatisfactory cure rates, and alternative therapeutic molecules are still required. However, their development has been hampered because accessible cell models supporting relevant HBV replication and appropriate antiviral activity are lacking. Strategies that reverse epigenetic alterations offer a unique opportunity for cell reprogramming, which is valuable for restoring altered cellular functions in human cell lines. This work aimed to investigate the feasibility of converting HepG2 cells that stably overexpress the HBV entry receptor (sodium/taurocholate cotransporting polypeptide, NTCP) toward IFNα-responsive cells using epigenetic reprogramming. Herein, we showed that an epigenetic regimen with non-cytotoxic doses of the demethylating compound 5-azacytidine restored the anti-HBV action of IFNα in epigenetically reprogrammed HepG2-NTCP-C4 cells, named REP-HepG2-NTCP cells. Thus, a significant inhibition in HBV DNA levels was measured in REP-HepG2-NTCP cells after IFNα treatment. This inhibitory effect was associated with the enhancement of IFNα-mediated induction of critical interferon-stimulated genes (ISGs), which was limited in non-reprogrammed cells. In particular, our data indicated that re-expression of 2'-5'-oligoadenylate synthetase 1 (OAS1) and interferon regulatory factor 9 (IRF9) was the result of an epigenetically driven unmasking of these genes in reprogrammed cells. At last, we evaluated the therapeutic potential of the IFN analog CDM-3008 in REP-HepG2-NTCP cells and demonstrated the efficiency of this chemical compound in triggering ISG induction and HBV inhibition. In summary, this study shows that epigenetic reprogramming promotes the IFNα response in HBV-infected cells and is potentially attractive for cell-based experimental screening of IFN-like compounds.
慢性乙型肝炎病毒（HBV）感染仍然是一个影响全球约2.5亿人的健康负担。到目前为止，现有的基于α干扰素（IFNα）的治疗方法治愈率并不理想，仍然需要替代治疗分子。然而，由于缺乏支持相关HBV复制和适当抗病毒活性的可及细胞模型，它们的开发受到了阻碍。逆转表观遗传改变的策略为细胞重编程提供了独特的机会，这对于恢复人类细胞系中改变的细胞功能很有价值。这项工作旨在研究使用表观遗传重编程将稳定过表达HBV进入受体（钠/牛磺胆酸共转运多肽，NTCP）的HepG2细胞转化为对IFNα有反应的细胞的可行性。在此，我们表明，用非细胞毒性剂量的去甲基化化合物5-氮杂胞苷进行的表观遗传方案恢复了IFNα在表观遗传重编程的HepG2-NTCP-C4细胞（称为REP-HepG2-NTCP细胞）中的抗HBV作用。因此，在IFNα处理后，在REP-HepG2-NTCP细胞中检测到HBV DNA水平有显著抑制。这种抑制作用与IFNα介导的关键干扰素刺激基因（ISG）诱导的增强有关，而在未重编程的细胞中这种诱导是有限的。特别是，我们的数据表明重新表达2'-5'-寡腺苷酸合成酶1（OAS1）和干扰素调节因子9（IRF9）是这些基因在重编程细胞中表观遗传驱动的去沉默的结果。最后，我们评估了IFN类似物CDM-3008在REP-HepG2-NTCP细胞中的治疗潜力，并证明了这种化合物在触发ISG诱导和HBV抑制方面的有效性。总之，这项研究表明表观遗传重编程促进了HBV感染细胞中的IFNα反应，并且对于基于细胞的IFN样化合物实验筛选可能具有吸引力。
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