Division of Dengue Hemorrhagic Fever Research, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
PLoS Negl Trop Dis. 2020 Nov 20;14(11):e0008835. doi: 10.1371/journal.pntd.0008835. eCollection 2020 Nov.
Suitable cell models are essential to advance our understanding of the pathogenesis of liver diseases and the development of therapeutic strategies. Primary human hepatocytes (PHHs), the most ideal hepatic model, are commercially available, but they are expensive and vary from lot-to-lot which confounds their utility. We have recently developed an immortalized hepatocyte-like cell line (imHC) from human mesenchymal stem cells, and tested it for use as a substitute model for hepatotropic infectious diseases. With a special interest in liver pathogenesis of viral infection, herein we determined the suitability of imHC as a host cell target for dengue virus (DENV) and as a model for anti-viral drug testing. We characterized the kinetics of DENV production, cellular responses to DENV infection (apoptosis, cytokine production and lipid droplet metabolism), and examined anti-viral drug effects in imHC cells with comparisons to the commonly used hepatoma cell lines (HepG2 and Huh-7) and PHHs. Our results showed that imHC cells had higher efficiencies in DENV replication and NS1 secretion as compared to HepG2 and Huh-7 cells. The kinetics of DENV infection in imHC cells showed a slower rate of apoptosis than the hepatoma cell lines and a certain similarity of cytokine profiles to PHHs. In imHC, DENV-induced alterations in levels of lipid droplets and triacylglycerols, a major component of lipid droplets, were more apparent than in hepatoma cell lines, suggesting active lipid metabolism in imHC. Significantly, responses to drugs with DENV inhibitory effects were greater in imHC cells than in HepG2 and Huh-7 cells. In conclusion, our findings suggest superior suitability of imHC as a new hepatocyte model for studying mechanisms underlying viral pathogenesis, liver diseases and drug effects.
合适的细胞模型对于深入了解肝脏疾病的发病机制和开发治疗策略至关重要。原代人肝细胞(PHH)是最理想的肝模型,但它们价格昂贵,且批次间存在差异,这限制了其应用。我们最近从人间充质干细胞中开发了一种永生化的肝样细胞系(imHC),并测试其作为肝嗜性传染病替代模型的适用性。鉴于对病毒感染导致的肝脏发病机制的特别关注,本文旨在确定 imHC 作为登革热病毒(DENV)宿主细胞靶标和抗病毒药物检测模型的适用性。我们研究了 DENV 产生的动力学、细胞对 DENV 感染的反应(细胞凋亡、细胞因子产生和脂滴代谢),并比较了 imHC 细胞与常用肝癌细胞系(HepG2 和 Huh-7)和 PHH 对抗病毒药物的作用。结果表明,与 HepG2 和 Huh-7 细胞相比,imHC 细胞在 DENV 复制和 NS1 分泌方面效率更高。imHC 细胞中 DENV 感染的动力学显示,细胞凋亡的速度比肝癌细胞系更慢,与 PHH 的细胞因子谱具有一定的相似性。在 imHC 细胞中,DENV 诱导的脂滴水平和三酰基甘油(脂滴的主要成分)变化比肝癌细胞系更明显,提示 imHC 细胞中存在活跃的脂代谢。重要的是,imHC 细胞对具有抑制 DENV 作用的药物的反应大于 HepG2 和 Huh-7 细胞。综上所述,我们的研究结果表明,imHC 作为研究病毒发病机制、肝脏疾病和药物作用的新型肝细胞模型具有更好的适用性。