RNA-Seq Revealed a Circular RNA-microRNA-mRNA Regulatory Network in Hantaan Virus Infection
RNA-Seq Revealed a Circular RNA-microRNA-mRNA Regulatory Network in Hantaan Virus Infection
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DOI：
10.3389/fcimb.2020.00097
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发表时间：
2020-03-13
影响因子：
5.7
通讯作者：
Hou Wei
中科院分区：
文献类型：
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作者：
Lu Shuang;Zhu Ni;Hou Wei
汉滩病毒(Hantaan Virus，HTNV)是一种流行于亚洲的汉坦病毒，可引起人类高死亡率的肾综合征出血热(HFRS)。然而，HTNV感染的发病机制仍不清楚。环状RNA(CircRNAs)是一种新型的非编码RNA，在多种致病过程中发挥着重要作用。然而，CircRNA的表达谱及其在HTNV感染发病机制中的作用仍是完全未知的。本研究采用RNA测序技术分析HTNV感染和模拟感染的人脐静脉内皮细胞(HUVECs)的CircRNA、microRNA(MiRNA)和mRNA表达谱。共有70个CircRNAs、66个miRNAs和788个mRNAs差异表达。利用RT-qPCR对差异表达的RNA进行验证。此外，我们还验证了一些差异表达的RNA，如CIRC_0000479、miR-149-5p、miR-330-5p、miR-411-3p、RIG-I、CMPK2、PARP10和GBP1，促进或抑制了HTNV的复制。基因本体论(GO)和京都基因与基因组百科全书(KEGG)的丰富分析表明，差异表达的CircRNAs的宿主基因主要参与天然免疫反应、I型干扰素信号通路和细胞因子介导的信号通路。此外，对CircRNA-miRNA-mRNA调控网络进行了整体分析。结果表明，HTNV感染过程中存在多种CircRNA-miRNA-mRNA相互作用。通过双荧光素酶报告基因检测，我们证实CIRC_0000479通过海绵化miR-149-5p间接调节RIG-I的表达，抑制病毒复制。这项研究首次对HTNV诱导的CircRNAs进行了全面的综述，揭示了丰富的CircRNAs和CircRNA相关的竞争性内源性RNAs(CeRNAs)参与了HTNV感染的调控，从而为HTNV与宿主相互作用的机制提供了新的见解。
Hantaan virus (HTNV), a Hantavirus serotype that is prevalent in Asia, causes hemorrhagic fever with renal syndrome (HFRS) with high mortality in human race. However, the pathogenesis of HTNV infection remains elusive. Circular RNAs (circRNAs), a new type of non-coding RNAs, play a crucial role in various pathogenic processes. Nevertheless, circRNA expression profiles and their effects on pathogenesis of HTNV infection are still completely unknown. In the present study, RNA sequencing was performed to analyze the circRNA, microRNA (miRNA), and mRNA expression profiles in HTNV-infected and mock-infected human umbilical vein endothelial cells (HUVECs). A total of 70 circRNAs, 66 miRNAs, and 788 mRNAs were differently expressed. Several differentially expressed RNAs were validated by RT-qPCR. Moreover, we verified that some differentially expressed RNAs, such as circ_0000479, miR-149-5p, miR-330-5p, miR-411-3p, RIG-I, CMPK2, PARP10, and GBP1, promoted or inhibited HTNV replication. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis demonstrated that the host genes of differentially expressed circRNAs were principally involved in the innate immune response, the type I interferon (IFN) signaling pathway, and the cytokine-mediated signaling pathway. Additionally, the circRNA-miRNA-mRNA regulatory network was integrally analyzed. The data showed that there were many circRNA-miRNA-mRNA interactions in HTNV infection. By dual-luciferase reporter assay, we confirmed that circ_0000479 indirectly regulated RIG-I expression by sponging miR-149-5p, hampering viral replication. This study for the first time presents a comprehensive overview of circRNAs induced by HTNV and reveals that a network of enriched circRNAs and circRNA-associated competitive endogenous RNAs (ceRNAs) is involved in the regulation of HTNV infection, thus offering new insight into the mechanisms underlying HTNV-host interaction.