Department of Gastroenterology and Hepatology, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Laboratory of Proteome Research/Proteome for Drug Discovery, National Institute of Biomedical Innovation, Health and Nutrition, 7-6-8, Saito-Asagi, Ibaraki, Osaka, 567-0085, Japan.
Sci Rep. 2020 Sep 17;10(1):15290. doi: 10.1038/s41598-020-72294-y.
Autophagy, a degradation system, works to maintain cellular homeostasis. However, as the impact of Hepatitis C virus (HCV) infection on hepatocyte autophagy and its effect on HCV replication remain unclear, we examined them. HCV infection suppressed late-stage autophagy and increased Rubicon. siRNA-mediated knockdown of Rubicon promoted autophagy in HCV-infected cells. In Huh-7 cells harbouring the HCV replicon, Rubicon knockdown downregulated the expression of type 1 interferon (IFN)-related genes and upregulated HCV replication. Rubicon overexpression or administration of bafilomycin A1 or chloroquine, an inhibitor of late-stage autophagy, suppressed autophagy and activated the type 1 IFN pathway. On the other hand, Atg7 knockout suppressed early-stage autophagy and did not activate the type 1 IFN pathway. In livers of humanized liver chimeric mice, HCV infection increased Rubicon and enhanced type 1 IFN signalling. Elimination of HCV in the mice reduced the increase in Rubicon due to HCV infection. The expression levels of Rubicon and IFN-stimulated genes in chronic hepatitis C patients were higher than those in non-B, non-C hepatitis patients. HCV infection increased Rubicon and suppressed hepatocyte autophagy, leading to activation of the intracellular immune response. Rubicon induction is involved in HCV replication via activation of the intracellular immune response.
自噬是一种降解系统,其作用是维持细胞内环境稳定。然而,丙型肝炎病毒(HCV)感染对肝细胞自噬的影响及其对 HCV 复制的影响尚不清楚,因此我们对此进行了研究。HCV 感染抑制晚期自噬并增加 Rubicon。Rubicon 的 siRNA 介导的敲低促进 HCV 感染细胞中的自噬。在含有 HCV 复制子的 Huh-7 细胞中,Rubicon 敲低下调了 I 型干扰素(IFN)相关基因的表达并上调了 HCV 复制。Rubicon 过表达或用巴弗洛霉素 A1 或氯喹(晚期自噬的抑制剂)处理可抑制自噬并激活 I 型 IFN 途径。另一方面,Atg7 敲除抑制早期自噬,但不会激活 I 型 IFN 途径。在人源化肝嵌合小鼠的肝脏中,HCV 感染增加了 Rubicon 并增强了 I 型 IFN 信号转导。小鼠中 HCV 的消除减少了由于 HCV 感染而增加的 Rubicon。慢性丙型肝炎患者的 Rubicon 和 IFN 刺激基因的表达水平高于非 B、非 C 肝炎患者。HCV 感染增加了 Rubicon 并抑制了肝细胞自噬,导致细胞内免疫反应的激活。Rubicon 的诱导通过激活细胞内免疫反应参与 HCV 复制。