MiR-10a-5p-Mediated Syndecan 1 Suppression Restricts Porcine Hemagglutinating Encephalomyelitis Virus Replication
MiR-10a-5p-Mediated Syndecan 1 Suppression Restricts Porcine Hemagglutinating Encephalomyelitis Virus Replication
复制标题
MiR-10a-5p 介导的 Syndecan 1 抑制限制猪血凝性脑脊髓炎病毒复制
DOI：
10.3389/fmicb.2020.00105
复制
发表时间：
2020-02-20
影响因子：
5.2
通讯作者：
He, Wenqi
中科院分区：
文献类型：
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作者：
Hu, Shiyu;Li, Zi;He, Wenqi
猪血凝性脑脊髓炎病毒（PHEV）是一种单链RNA冠状病毒，可引起感染宿主的神经功能障碍，并通过调节特定的microRNA（miRNA）水平导致宿主转录组的广泛改变。miRNA通过促进抗病毒免疫应答、增强病毒复制或改变miRNA介导的宿主基因调控而参与RNA病毒的发病机制。因此，探索PHEV感染宿主中发生的病毒-miRNA相互作用可能导致识别对抗病毒生命周期或发病机制的新机制。在此，我们发现PHEV在体外培养的N2 a细胞和体内培养的小鼠脑组织中均能组成性上调miR-10a-5 p的表达。用miR-10a-5 p模拟物处理允许miR-10a-5 p富集并导致PHEV复制的显著限制，表明miR-10a-5 p对RNA病毒感染的广泛负调控。结果也通过miR-10a-5 p抑制剂过表达得到证实。荧光素酶报告基因、实时定量PCR（qRT-PCR）和蛋白质印迹分析进一步表明，Syndecan 1（SDC 1）是一种与宿主防御机制相关的细胞表面蛋白聚糖，在PHEV感染期间充当miR-10a-5 p的靶基因。自然地，siRNA介导的SDC 1敲低导致病毒复制减少，这意味着SDC 1表达可能是病毒复制的有利条件。总而言之，研究结果表明，丰富的miR-10a-5 p导致了SDC 1的下游抑制，并且它在PHEV诱导的疾病中发挥着抗病毒机制的作用，为未来预防和治疗PHEV感染提供了潜在的策略。工作。
Porcine hemagglutinating encephalomyelitis virus (PHEV) is a single-stranded RNA coronavirus that causes nervous dysfunction in the infected hosts and leads to widespread alterations in the host transcriptome by modulating specific microRNA (miRNA) levels. MiRNAs contribute to RNA virus pathogenesis by promoting antiviral immune response, enhancing viral replication, or altering miRNA-mediated host gene regulation. Thus, exploration of the virus–miRNA interactions occurring in PHEV-infected host may lead to the identification of novel mechanisms combating the virus life cycle or pathogenesis. Here, we discovered that the expression of miR-10a-5p was constitutively up-regulated by PHEV in both the N2a cells in vitro and mice brain in vivo. Treatment with miR-10a-5p mimics allowed miR-10a-5p enrichment and resulted in a significant restriction in PHEV replication, suggesting widespread negative regulation of the RNA virus infection by miR-10a-5p. The outcomes were also evidenced by miR-10a-5p inhibitor over-expression. Luciferase reporter, quantitative real-time PCR (qRT-PCR), and western blotting analysis further showed that Syndecan 1 (SDC1), a cell surface proteoglycan associated with host defense mechanisms, acts as a target gene of miR-10a-5p during PHEV infection. Naturally, siRNA-mediated knockdown of SDC1 leads to a reduction in viral replication, implying that SDC1 expression is likely a favorable condition for viral replication. Together, the findings demonstrated that the abundant miR-10a-5p leads to downstream suppression of SDC1, and it functions as an antiviral mechanism in the PHEV-induced disease, providing a potential strategy for the prevention and treatment of PHEV infection in the future work.