宿主差异microRNA对猪繁殖与呼吸综合征病毒感染复制的调控机制解析

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically important pathogens in the global swine industry and its primary target cell is the porcine alveolar macrophage (PAMs). In recent years, there is a growing body of evidence that cellular microRNAs (miRNAs) are important regulators of innate immune response and the intricate networks of host-pathogen interactions. To identify the differential expression of host miRNAs and their roles in PRRSV infection and replication, this project conducts the following studies. (1) The host differential expressed miRNAs in PRRSV infection is measured by small RNA deep-sequencing. Then the miRNA-mRNA co-expressional networks are established by combining the data of differential expressed mRNAs. (2) The relationship between the host differential expressed miRNAs and PRRSV replication is verified. (3) The targets of miRNAs and miRNA-mediated signaling are investigated using dual-luciferase assays and RNA interference. By analyzing the impact of PRRSV infection on the host miRNA expression and clarifying the roles of host differential expressed miRNAs in PRRSV infection and replication from higher viewpoint and wider horizons, our project is of great significance for the further explore of miRNA-virus interations.

猪繁殖与呼吸综合征病毒（PRRSV）是危害全球养猪业的重要病原之一，其首要感染的靶细胞是肺泡巨噬细胞（PAMs）。宿主细胞microRNA（miRNA）是机体先天性免疫及宿主-病原体互作网络中不可或缺的调节因子。为了探明宿主差异miRNA对PRRSV感染复制的调控机制，本项目拟进行以下研究：（1）利用深度测序和统计分析筛选PRRSV感染PAMs的差异miRNA，结合下游差异mRNA构建miRNA-mRNA共表达网络；（2）利用过表达或沉默技术验证差异miRNA与PRRSV复制的作用关系；（3）利用双荧光素酶系统和RNA干扰等技术探明差异miRNA作用的靶基因及相关通路的调控机制。该研究结果通过解析PRRSV感染对宿主miRNA表达谱的影响，从更全面的角度阐释宿主差异miRNA在PRRSV感染复制过程中的不同角色，对揭示宿主miRNA与病毒相互作用的机制研究方面有重要意义。

猪繁殖与呼吸综合征病毒（PRRSV）是危害全球养猪业的重要病原之一。宿主细胞microRNA（miRNA）是机体先天性免疫及宿主-病原体互作网络中不可或缺的调节因子。本项目解析了宿主差异miRNA对PRRSV感染复制的调控机制，得出以下结果：（1）利用深度测序获得PRRSV感染PAMs的差异miRNA，结合下游差异mRNA构建miRNA-mRNA共表达网络；（2）筛选出ssc-miR-9可通过调控其靶基因DDX46表达进而抑制PRRSV的感染和复制；（3）通过构建含有宿主内源miRNA结合序列的突变PRRSV并分析其生长特性，证明利用不同丰度的细胞内源miRNA可对PRRSV进行操纵和改造，并对PRRSV的复制产生抑制作用。该研究结果通过解析PRRSV感染对宿主miRNA表达谱的影响，从更全面的角度阐释宿主差异miRNA在PRRSV感染复制过程中的不同角色，对揭示宿主miRNA与病毒相互作用的机制研究方面有重要意义。