Table_7_Whole-transcriptome analyses of sheep embryonic testicular cells infected with the bluetongue virus.xlsx

Introductionbluetongue virus (BTV) infection triggers dramatic and complex changes in the host's transcriptional profile to favor its own survival and reproduction. However, there is no whole-transcriptome study of susceptible animal cells with BTV infection, which impedes the in-depth and systematical understanding of the comprehensive characterization of BTV-host interactome, as well as BTV infection and pathogenic mechanisms.
Methodsto systematically understand these changes, we performed whole-transcriptome sequencing in BTV serotype 1 (BTV-1)-infected and mock-infected sheep embryonic testicular cells, and subsequently conducted bioinformatics differential analyses.
Resultsthere were 1504 differentially expressed mRNAs, 78 differentially expressed microRNAs, 872 differentially expressed long non-coding RNAs, and 59 differentially expressed circular RNAs identified in total. Annotation from the Gene Ontology, enrichment from the Kyoto Encyclopedia of Genes and Genomes, and construction of competing endogenous RNA networks revealed differentially expressed RNAs primarily related to virus-sensing and signaling transduction pathways, antiviral and immune responses, inflammation, and development and metabolism related pathways. Furthermore, a protein-protein interaction network analysis found that BTV may contribute to abnormal spermatogenesis by reducing steroid biosynthesis. Finally, real-time quantitative PCR and western blotting results showed that the expression trends of differentially expressed RNAs were consistent with the whole-transcriptome sequencing data.
Discussionthis study provides more insights of comprehensive characterization of BTV-host interactome, and BTV infection and pathogenic mechanisms.

引言：蓝舌病毒（bluetongue virus, BTV）感染会引发宿主转录组发生剧烈且复杂的改变，以助力自身的存活与增殖。然而目前尚无针对BTV感染易感动物细胞的全转录组研究，这阻碍了我们对BTV与宿主互作组的全面解析，以及BTV感染与致病机制的深入系统理解。
方法：为系统解析此类转录组改变，本研究对感染1型蓝舌病毒（BTV serotype 1, BTV-1）的绵羊胚胎睾丸细胞以及模拟感染的对照细胞开展了全转录组测序，并随后进行了生物信息学差异分析。
结果：本研究共鉴定得到1504个差异表达mRNA、78个差异表达微小RNA（microRNA）、872个差异表达长链非编码RNA以及59个差异表达环状RNA。通过基因本体论（Gene Ontology, GO）注释、京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）富集分析以及内源竞争RNA（competing endogenous RNA, ceRNA）网络构建，发现差异表达RNA主要富集于病毒识别与信号转导通路、抗病毒与免疫应答、炎症反应以及发育与代谢相关通路。进一步的蛋白质相互作用网络分析显示，BTV可能通过降低类固醇生物合成途径的活性，引发精子发生异常。最后，实时定量聚合酶链反应（real-time quantitative PCR, qPCR）与蛋白质免疫印迹（western blotting, WB）实验结果证实，差异表达RNA的表达趋势与全转录组测序数据一致。
讨论：本研究为BTV与宿主互作组的全面解析，以及BTV感染与致病机制的研究提供了更多新的见解。