MicroRNA Regulation of Human Genes Essential for Influenza A (H7N9) Replication

• Published in: PloS one 2016-05, Vol.11 (5), p.e0155104-e0155104
• Main Authors: Wolf, Stefan, Wu, Weilin, Jones, Cheryl, Perwitasari, Olivia, Mahalingam, Suresh, Tripp, Ralph A
• Format: Article
• Language: English
• Subjects: A549 Cells; Animals; Antiviral agents; Antiviral Agents - metabolism; Avian flu; Biology and life sciences; Cytokines; Dogs; Drug resistance; Gene expression; Gene Expression Regulation; Gene Knockdown Techniques; Gene regulation; Genes; Genetic aspects; Genetic regulation; Genome, Human; Genomes; Health aspects; Hepatitis; High-throughput screening; High-Throughput Screening Assays; Humans; Immunology; Infections; Infectious diseases; Inflammation; Influenza; Influenza A; Influenza A virus; Influenza A Virus, H7N9 Subtype - physiology; Influenza viruses; Influenza, Human - genetics; Influenza, Human - virology; Kinases; Leukemia; Leukemia Inhibitory Factor - metabolism; Madin Darby Canine Kidney Cells; Medicine and health sciences; MicroRNA; MicroRNAs; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; Models, Biological; Morbidity; Mortality; Pandemics; Pathogenesis; Properties; Public health; Replication; Reproducibility of Results; Respiratory diseases; Ribonucleic acid; RNA; siRNA; Software; Strains (organisms); Target recognition; Viral infections; Virus replication; Virus Replication - physiology; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0155104

Influenza A viruses are important pathogens of humans and animals. While seasonal influenza viruses infect humans every year, occasionally animal-origin viruses emerge to cause pandemics with significantly higher morbidity and mortality rates. In March 2013, the public health authorities of China reported three cases of laboratory confirmed human infection with avian influenza A (H7N9) virus, and subsequently there have been many cases reported across South East Asia and recently in North America. Most patients experience severe respiratory illness, and morbidity with mortality rates near 40%. No vaccine is currently available and the use of antivirals is complicated due the frequent emergence of drug resistant strains. Thus, there is an imminent need to identify new drug targets for therapeutic intervention. In the current study, a high-throughput screening (HTS) assay was performed using microRNA (miRNA) inhibitors to identify new host miRNA targets that reduce influenza H7N9 replication in human respiratory (A549) cells. Validation studies lead to a top hit, hsa-miR-664a-3p, that had potent antiviral effects in reducing H7N9 replication (TCID50 titers) by two logs. In silico pathway analysis revealed that this microRNA targeted the LIF and NEK7 genes with effects on pro-inflammatory factors. In follow up studies using siRNAs, anti-viral properties were shown for LIF. Furthermore, inhibition of hsa-miR-664a-3p also reduced virus replication of pandemic influenza A strains H1N1 and H3N2.