Identification and analysis of differentially-expressed microRNAs in Japanese encephalitis virus-infected PK-15 cells with deep sequencing.

Yuhan Cai,Qiaoli Lang,Xinqiong Li,Ling Zhu,Xiaowan Liu,Xiao Liu,Zhiwen Xu,Yuanchen Zhou,Xiaogai Qiao

doi:10.3390/ijms16012204

Abstract

Japanese encephalitis virus (JEV), a mosquito-borne Flavivirus, causes acute viral encephalitis with high morbidity and mortality in humans and animals. MicroRNAs (miRNAs) are small noncoding RNAs that are important modulators of the intricate host-pathogen interaction networks. However, our knowledge of the changes that occur in miRNAs in host cells after JEV infection is still limited. To understand the molecular pathogenesis of JEV at the level of posttranscriptional regulation, we used Illumina deep sequencing to sequence two small RNA libraries prepared from PK-15 cells before and after JEV infection. We identified 522 and 427 miRNAs in the infected and uninfected cells, respectively. Overall, 132 miRNAs were expressed significantly differently after challenge with JEV: 78 were upregulated and 54 downregulated. The sequencing results for selected miRNAs were confirmed with RT-qPCR. GO analysis of the host target genes revealed that these dysregulated miRNAs are involved in complex cellular pathways, including the metabolic pathway, inflammatory response and immune response. To our knowledge, this is the first report of the comparative expression of miRNAs in PK-15 cells after JEV infection. Our findings will underpin further studies of miRNAs’ roles in JEV replication and identify potential candidates for antiviral therapies against JEV.

Highlights

Japanese encephalitis virus (JEV), a mosquito-borne member of the genus Flavivirus, is the most important causative agent of acute viral encephalitis in humans, occurring mainly in Asian countries [1].Approximately 50,000 cases of JEV occur per year worldwide, resulting in more than 10,000 deaths and 15,000 cases of neurological or psychiatric sequelae [2]
PK-15 cells are a good model in which to evaluate the role of cellular miRNAs in the host response to JEV infection
Using deep sequencing, we have described for the first time a set of miRNAs dysregulated in PK-15 cells by JEV infection

Summary

Introduction

Japanese encephalitis virus (JEV), a mosquito-borne member of the genus Flavivirus, is the most important causative agent of acute viral encephalitis in humans, occurring mainly in Asian countries [1]. 50,000 cases of JEV occur per year worldwide, resulting in more than 10,000 deaths and 15,000 cases of neurological or psychiatric sequelae [2]. The actual number of JEV infections is probably much higher, possibly up to 175,000 annually [3]. JEV is one of the main causes of infectious reproductive failure in swine, resulting in significant economic losses in the swine industry [1,4]. To effectively control JEV infection, it is essential to clarify the mechanisms of virus pathogenesis

Objectives

Results

Conclusion