Deep Sequencing Identified miR-193b-3p as a Positive Regulator of Autophagy Targeting Akt3 in Ctenopharyngodon idella CIK Cells During GCRV Infection

Hongyan Yu; Zheyan Chen; Yuting Liu; Yubang Shen; Lang Gui; Junqiang Qiu; Xiaoyan Xu; Jiale Li

doi:10.1016/j.fsi.2024.109586

Deep Sequencing Identified miR-193b-3p as a Positive Regulator of Autophagy Targeting Akt3 in Ctenopharyngodon idella CIK Cells During GCRV Infection

Fish Shellfish Immunol. 2024 Apr 24:109586. doi: 10.1016/j.fsi.2024.109586. Online ahead of print.

Authors

Hongyan Yu¹, Zheyan Chen¹, Yuting Liu¹, Yubang Shen¹, Lang Gui¹, Junqiang Qiu¹, Xiaoyan Xu², Jiale Li³

Affiliations

¹ Key Laboratory of Freshwater Aquatic Genetic Resources Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, China; Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai, China.
² Key Laboratory of Freshwater Aquatic Genetic Resources Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, China; Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai, China; National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai, China. Electronic address: xyxu@shou.edu.cn.
³ Key Laboratory of Freshwater Aquatic Genetic Resources Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, China; Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai, China; National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai, China. Electronic address: jlli@shou.edu.cn.

PMID: 38670410
DOI: 10.1016/j.fsi.2024.109586

Abstract

Recent research has highlighted complex and close interaction between miRNAs, autophagy, and viral infection. In this study, we observed the autophagy status in CIK cells infected with GCRV at various time points. We found that GCRV consistently induced cellar autophagy from 0 h to 12 h post infection. Subsequently, we performed deep sequencing on CIK cells infected with GCRV at 0 h and 12 h respectively, identifying 38 DEMs and predicting 9,581 target genes. With the functional enrichment analyses of GO and KEGG, we identified 35 autophagy-related target genes of these DEMs, among which akt3 was pinpointed as the most central hub gene using module assay of the PPI network. Then employing the miRanda and Targetscan programs for prediction, and verification through a double fluorescent enzyme system and qPCR method, we confirmed that miR-193b-3p could target the 3'-UTR of grass carp akt3, reducing its gene expression. Ultimately, we illustrated that grass carp miR-193b-3p could promote autophagy in CIK cells. Above results collectively indicated that miRNAs might play a critical role in autophagy of grass carp during GCRV infection and contributed significantly to antiviral immunity by targeting autophagy-related genes. This study may provide new insights into the intricate mechanisms involved in virus, autophagy, and miRNAs.

Keywords: GCRV; autophagy; grass carp; miRNAs.