Lv Lilei, Luo Huaye, Zhang Min, Wu Chuntao, Jiang Yifeng, Tong Wu, Li Guoxin, Zhou Yanjun, Li Yanhua, Wang Zhao, Liu Changlong
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, PR China.
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, PR China; Department of Laboratory Animal Sciences, School of Basic Medicine, Naval Medical University, Shanghai 200433, PR China.
Virus Res. 2024 Dec;350:199502. doi: 10.1016/j.virusres.2024.199502. Epub 2024 Nov 23.
Porcine epidemic diarrhea virus (PEDV) is a highly contagious virus that poses a serious threat to the global pig industry. Despite extensive efforts, the mechanism underlying virus entry for PEDV remains elusive. In this study, we first identified PEDV-susceptible and non-susceptible cell lines by using PEDV spike pseudotyped vesicular stomatitis virus. Subsequently, we conducted a comprehensive transcriptomic analysis on these cell lines. Through integrating differential expression gene analysis with weighted gene co-expression network analysis, we identified the key pathways that are correlated with the PEDV entry. Our analysis revealed a strong correlation between cholesterol, sterols, and lipid transport with PEDV entry, suggesting a potential role for cholesterol transport in the PEDV entry. For further investigation, we treated Huh7, Vero and LLC-PK1 cells with a cholesterol transport inhibitor, ezetimibe, and observed a significant inhibition of PEDV entry and subsequent viral replication in these cells. Interestingly, pre-treating Huh7 cells with ezetimibe resulted in an increase in the entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Middle East respiratory syndrome coronavirus (MERS-CoV) pseudoviruses. Moreover, we found that cholesterol could facilitate the entry of PEDV into Huh7 and Vero cells, and this promoting effect can be blocked by ezetimibe. These findings suggest that targeting cholesterol transport specifically inhibits PEDV entry into susceptible cells. Our study offers novel insights into the mechanism of PEDV entry and the development of new therapeutic strategies against this economically important virus.
猪流行性腹泻病毒（PEDV）是一种具有高度传染性的病毒，对全球养猪业构成严重威胁。尽管已付出巨大努力，但PEDV进入细胞的机制仍不清楚。在本研究中，我们首先使用PEDV刺突蛋白假型化的水疱性口炎病毒鉴定出对PEDV敏感和不敏感的细胞系。随后，我们对这些细胞系进行了全面的转录组分析。通过将差异表达基因分析与加权基因共表达网络分析相结合，我们确定了与PEDV进入相关的关键途径。我们的分析揭示了胆固醇、甾醇和脂质转运与PEDV进入之间存在强烈相关性，表明胆固醇转运在PEDV进入过程中可能发挥作用。为进一步研究，我们用胆固醇转运抑制剂依泽替米贝处理Huh7、Vero和LLC-PK1细胞，并观察到这些细胞中PEDV进入及随后的病毒复制受到显著抑制。有趣的是，用依泽替米贝预处理Huh7细胞会导致严重急性呼吸综合征冠状病毒2（SARS-CoV-2）和中东呼吸综合征冠状病毒（MERS-CoV）假病毒的进入增加。此外，我们发现胆固醇可促进PEDV进入Huh7和Vero细胞，且这种促进作用可被依泽替米贝阻断。这些发现表明，靶向胆固醇转运可特异性抑制PEDV进入易感细胞。我们的研究为PEDV进入机制以及针对这种具有重要经济意义的病毒开发新的治疗策略提供了新见解。
