mRNA profiling during infection with H1N1 influenza A virus (A/Mexico/InDRE4487/H1N1/2009). Homo sapiens

MicroRNAs (miRNAs) repress the expression levels of genes by binding to mRNA transcripts, acting as master regulators of cellular processes. Differential expression of miRNAs has been linked to viral-associated diseases involving members of the hepacivirus, herpesvirus, and retrovirus families. In contrast, limited biological and molecular information has been reported on the potential role of cellular miRNAs in the lifecycle of influenza A viruses (infA). In this study, we hypothesize that elucidating the miRNA expression signatures induced by low-pathogenic swine-origin influenza A virus (S-OIV) pandemic H1N1 (2009) and highly pathogenic avian-origin (A-OIV) H7N7 (2003) infections could reveal temporal and strain-specific miRNA fingerprints during the viral lifecycle, shedding important insights into the potential role of cellular miRNAs in host-infA interactions. Using a microfluidic microarray platform, we profiled cellular miRNA expression in human A549 cells infected with S- and A-OIVs at multiple time-points during the viral lifecycle, including global gene expression profiling during S-OIV infection. Using target prediction and pathway enrichment analyses, we identified the key cellular pathways associated with the differentially expressed miRNAs and predicted mRNA targets during infA infection, including immune system, cell proliferation, apoptosis, cell cycle, and DNA replication and repair. By identifying the specific and dynamic molecular phenotypic changes (microRNAome) triggered by S- and A-OIV infection in human cells, we provide experimental evidence demonstrating a series of temporal- and strain-specific host molecular responses involving different combinatorial contributions of multiple cellular miRNAs. Our results also identify novel potential exosomal miRNA biomarkers associated with pandemic S-OIV and deadly A-OIV-host infection. Overall design: Control (mock-infected) samples: 12 (2 technical replicates, averaged), Infected samples: 6, for each time-point (0, 4, 8, 24, 48 and 72 hours post infection). The experiment was performed in six replicates.

微小RNA（MicroRNAs, miRNAs）通过结合信使核糖核酸（mRNA）转录本抑制基因表达水平，作为细胞进程的核心调控因子。微小RNA的差异表达已被证实与肝炎病毒属、疱疹病毒科及逆转录病毒科成员引发的病毒相关疾病存在关联。与之相对，目前关于细胞内微小RNA在甲型流感病毒（influenza A viruses, infA）生命周期中潜在作用的生物学与分子机制研究报道较为有限。本研究提出假说：解析低致病性猪源甲型流感病毒（S-OIV）2009年大流行株H1N1与高致病性禽源甲型流感病毒（A-OIV）2003株H7N7感染所诱导的微小RNA表达特征，可揭示病毒生命周期中具有时间动态性与毒株特异性的微小RNA特征谱，为阐明细胞微小RNA在宿主-甲型流感病毒互作中的潜在作用提供重要见解。本研究采用微流控微阵列平台，对病毒生命周期多个时间点感染猪源与禽源甲型流感病毒的人A549细胞内细胞微小RNA表达谱进行分析，同时涵盖猪源甲型流感病毒感染期间的全基因表达谱分析。通过靶基因预测与通路富集分析，本研究鉴定出甲型流感病毒感染期间与差异表达微小RNA及预测的信使RNA靶标相关的关键细胞通路，包括免疫系统、细胞增殖、细胞凋亡、细胞周期以及DNA复制与修复通路。通过鉴定猪源与禽源甲型流感病毒感染人类细胞所引发的特异性、动态性分子表型变化（微小RNA组，microRNAome），本研究提供了实验证据，证明存在一系列具有时间动态性与毒株特异性的宿主分子应答，该应答涉及多种细胞微小RNA的不同组合调控。本研究结果还鉴定出与2009年大流行猪源甲型流感病毒及致死性禽源甲型流感病毒-宿主感染相关的新型潜在外泌体微小RNA生物标志物。实验整体设计：对照组（mock感染）样本共12份（取2次技术重复的平均值）；每个感染时间点（感染后0、4、8、24、48及72小时）的感染样本各6份。本实验共完成6次生物学重复。