Chen Yu, Liu Wen, Xu Haixu, Liu Jingjing, Deng Yonghuan, Cheng Hao, Zhu Shanshan, Pei Yuru, Hu Jiao, Hu Zenglei, Liu Xiaowen, Wang Xiaoquan, Gu Min, Hu Shunlin, Liu Xiufan
Animal Infectious Disease Laboratory, College of Veterinary Medicine, Yangzhou University, Yangzhou, China.
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis, College of Veterinary Medicine, Yangzhou University, Yangzhou, China.
Front Microbiol. 2019 Jul 23;10:1659. doi: 10.3389/fmicb.2019.01659. eCollection 2019.
Newcastle disease virus (NDV), causative agent of Newcastle disease (ND), is one of the most devastating pathogens for poultry industry worldwide. MicroRNAs (miRNAs) are non-coding RNAs that regulate gene expression by regulating mRNA translation efficiency or mRNA abundance through binding to mRNA directly. Accumulating evidence has revealed that cellular miRNAs can also affect virus replication by controlling host-virus interaction. To identify miRNA expression profile and explore the roles of miRNA during NDV replication, in this study, small RNA deep sequencing was performed of non-inoculated DF-1 cells (chicken embryo fibroblast cell line) and JS 5/05-infected cells collected at 6 and 12 h post infection (hereafter called mock' NDV-6 h, and NDV-12 h groups respectively). A total of 73 miRNAs of NDV-6 h group and 64miRNAs of NDV-12 h group were significantly differentially expressed (SDE) when compared with those in mock group. Meanwhile, 50 SDE miRNAs, including 48 up- and 2 down-regulated, showed the same expression patterns in NDV-6 h and NDV-12 h groups. qRT-PCR validation of 15 selected miRNAs' expression patterns was consistent with deep sequencing. To investigate the role of these SDE miRNAs in NDV replication, miRNA mimics and inhibitors were transfected into DF-1 cells followed by NDV infection. The results revealed that gga-miR-451 and gga-miR-199-5p promoted NDV replication while gga-miR-19b-3p and gga-miR-29a-3p inhibited NDV replication. Further function research demonstrated gga-miR-451 suppressed NDV-induced inflammatory response targeting YWHAZ (tyrosine3-monooxygenase/tryptophan5-monooxygenase activation protein zeta). Overall, our study presented a global miRNA expression profile in DF-1 cells in response to NDV infection and verified the roles of some SDE miRNAs in NDV replication which will underpin further studies of miRNAs' roles between the host and the virus.
新城疫病毒（NDV）是新城疫（ND）的病原体，是全球家禽业最具毁灭性的病原体之一。微小RNA（miRNA）是非编码RNA，通过直接与mRNA结合来调节mRNA翻译效率或mRNA丰度，从而调控基因表达。越来越多的证据表明，细胞miRNA也可以通过控制宿主与病毒的相互作用来影响病毒复制。为了鉴定miRNA表达谱并探索miRNA在NDV复制过程中的作用，在本研究中，对未接种的DF-1细胞（鸡胚成纤维细胞系）以及感染后6小时和12小时收集的JS 5/05感染细胞（以下分别称为mock、NDV-6 h和NDV-12 h组）进行了小RNA深度测序。与mock组相比，NDV-6 h组共有73个miRNA和NDV-12 h组共有64个miRNA显著差异表达（SDE）。同时，50个SDE miRNA，包括48个上调和2个下调的miRNA，在NDV-6 h和NDV-12 h组中表现出相同的表达模式。对15个选定miRNA表达模式的qRT-PCR验证与深度测序结果一致。为了研究这些SDE miRNA在NDV复制中的作用，将miRNA模拟物和抑制剂转染到DF-1细胞中，随后进行NDV感染。结果显示，gga-miR-451和gga-miR-199-5p促进NDV复制，而gga-miR-19b-3p和gga-miR-29a-3p抑制NDV复制。进一步的功能研究表明，gga-miR-451通过靶向YWHAZ（酪氨酸3-单加氧酶/色氨酸5-单加氧酶激活蛋白ζ）抑制NDV诱导的炎症反应。总体而言，我们的研究展示了DF-1细胞中响应NDV感染的全局miRNA表达谱，并验证了一些SDE miRNA在NDV复制中的作用，这将为进一步研究miRNA在宿主与病毒之间的作用奠定基础。
