Wu Ning, Yang Bo, Wen Bo, Wang Ting, Guo Jiaona, Qi Xuefeng, Wang Jingyu
College of Veterinary Medicine, Northwest A&F University, Xianyang, China.
Shanxi Academy of Advanced Research and Innovation, Taiyuan, China.
Front Microbiol. 2020 May 19;11:831. doi: 10.3389/fmicb.2020.00831. eCollection 2020.
Hydropericardium-hepatitis syndrome (HHS) is caused by some strains of fowl adenovirus serotype 4 (FAdV-4). However, the mechanism of FAdV-4 entry is not well understood. Therefore, to investigate the changes in host cellular response at the early stage of FAdV-4 infection, a conjoint analysis of miRNA-seq and mRNA-seq was utilized with leghorn male hepatocellular (LMH) cells at 30, 60, and 120 min after FAdV-4 infection. In total, we identified 785 differentially expressed (DE) miRNAs and 725 DE mRNAs in FAdV-4-infected LMH cells. Most miRNAs and mRNAs, including gga-miR-148a-3p, gga-miR-148a-5p, gga-miR-15c-3p, CRK, SOCS3, and EGR1, have not previously been reported to be associated with FAdV-4 infection. The conjoint analysis of the obtained data identified 856 miRNA-mRNA pairs at three time points. The interaction network analysis showed that gga-miR-128-2-5p, gga-miR-7475-5p, novel_miR205, and TCF7L1 were located in the core of the network. Furthermore, the relationship between gga-miR-128-2-5p and its target OBSL1 was confirmed using a dual-luciferase reporter system and a real-time quantitative polymerase chain reaction assay. experiments revealed that both gga-miR-128-2-5p overexpression and OBSL1 loss of function inhibited FAdV-4 entry. These results suggested that gga-miR-128-2-5p plays an important role in FAdV-4 entry by targeting OBSL1. To the best of our knowledge, the present study is the first to analyze host miRNA and mRNA expression at the early stage of FAdV-4 infection; furthermore, the results of this study help to elucidate the molecular mechanisms of FAdV-4 entry.
心包积水-肝炎综合征（HHS）由某些4型禽腺病毒（FAdV-4）毒株引起。然而，FAdV-4的进入机制尚不清楚。因此，为了研究FAdV-4感染早期宿主细胞反应的变化，在FAdV-4感染后30、60和120分钟，利用来航鸡雄性肝细胞（LMH）细胞对miRNA-seq和mRNA-seq进行了联合分析。我们总共在FAdV-4感染的LMH细胞中鉴定出785个差异表达（DE）的miRNA和725个DE mRNA。大多数miRNA和mRNA，包括gga-miR-148a-3p、gga-miR-148a-5p、gga-miR-15c-3p、CRK、SOCS3和EGR1，此前尚未报道与FAdV-4感染有关。对获得的数据进行联合分析，在三个时间点鉴定出856个miRNA-mRNA对。相互作用网络分析表明，gga-miR-128-2-5p、gga-miR-7475-5p、novel_miR205和TCF7L1位于网络核心。此外，使用双荧光素酶报告系统和实时定量聚合酶链反应测定法证实了gga-miR-128-2-5p与其靶标OBSL1之间的关系。实验表明，gga-miR-128-2-5p的过表达和OBSL1功能的丧失均抑制FAdV-4的进入。这些结果表明，gga-miR-128-2-5p通过靶向OBSL1在FAdV-4进入中起重要作用。据我们所知，本研究是首次分析FAdV-4感染早期宿主miRNA和mRNA的表达；此外，本研究结果有助于阐明FAdV-4进入的分子机制。
