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Abstract
Influenza virus causes mild to severe respiratory tract infections and is a substantial health concern causing seasonal epidemics and sporadic pandemics resulting in morbidity, mortality, and economic losses worldwide. Several anti-influenza drugs are available, but these agents target viral components and are susceptible to drug resistance. Influenza viruses require host genes to replicate. RNA interference (RNAi) screens can identify specific host genes coopted by influenza for replication. Targeting these pro-influenza genes can provide therapeutic strategies to reduce virus replication. Identifying microRNAs (miRs) that regulate these host genes can support an antiviral strategy while repurposing of clinically approved drugs that target cellular machinery necessary for influenza virus replication can provide a therapeutic approach for inhibiting influenza virus replication. Herein, we used RNA interference screening to identify key host cell genes required for influenza replication using human lung (A549) cells and identified 19 pro-influenza GPCR and 13 pro-influenza ion channel genes using small inferring RNAs (siRNA). These pro-influenza genes were authenticated by infecting siRNA transfected cells with A/WSN/33, A/CA/04/09, and B/Yamagata/16/1988 resulting in validation of 16 pro-influenza GPCR and 5 pro-influenza ion channel genes. These findings showed that several GPCR and ion channel genes are needed for production of infectious influenza virus and provide potential targets for the development of host-directed therapeutic strategies to impede the influenza productive cycle to limit infection. Thirty-three miRs predicted to target these pro-influenza host GPCR or ion channel genes were screened using A/WSN/33 A/CA/04/09, or B/Yamagata/16/1988 infected A549 cells using miR mimics and their paired inhibitors and evaluated for their effect on influenza replication. Several miRs prevented influenza virus replication through GPCR or ion channel gene regulation and 4 pan-anti-influenza miRs were identified. There is a need for new antiviral drug strategies that include repurposing of clinically approved drugs. Herein, we identified the FDA-approved drugs Clopidogrel and Triamterene and evaluated these drugs’ ability to inhibit A/WSN/33 and A/CA/04/09 influenza A strains, and Yamagata/16/1988 influenza B replication in vitro. Both Clopidogrel and Triamterene reduced influenza replication and spread across multiple strains and subtypes providing a potential druggable approach to influenza treatment.