Omics Method Screening Of Repurposing Drugs For Highly Pathogenic Avian Influenza Virus Infection And Immune Response And Lung Injury In COVID-19 Patients

Influenza viruses,which can cause seasonal epidemics and occasionally pandemics,posing a serious threat to human health.Highly pathogenic avian influenza(HPAI)viruses are usually distributed in birds,and occasionally infect humans and cause serious respiratory diseases,such as acute lung injury(ALI).According to the report of the World Health Organization,by the end of 2018,860 H5N1 patients were diagnosed,with a mortality rate of 52.79%.Although damage was caused by HPAI,there are limited drugs and treatments.In this study,we focused on exploring the pathogenesis of HPAI and potential therapeutic drugs.Drug repositioning is an economical way for drug discovery.In order to discover effective drugs to ameliorate HPAI H5N1-induced ALI,we explored new screening strategies for drug repositioning based on whole-genome RNAi and high-throughput transcriptome sequencing.First,we performed RNAi screening of 19,424 genes in A549 lung epithelial cells and examined cell viability following H5N1 infection.In total,we identified 1,137 host genes for which knockdown significantly altered cell viability by more than 20%.Searching these 1,137 genes in DrugBank,we obtained 104 commercially available drugs targeting 65 validated genes.We found 28 drugs that could significantly recover cell viability following H5N1 infection and tested the efficacy of several of these drugs in an H5N1-infection mouse model.The anticancer drug flavopiridol significantly reduced the leukocyte infiltration and lung injury score in H5N1-infected mice,improved the edema in the lung tissue of infected mice,and increased the survival rate of H5N1 infected mice.These results suggest that in addition to proven indications,flavopiridol might be a novel remedy for potential H5N1 epidemics.Moreover,we also identified several drugs as potential treatments for avian influenza A H5N1 virus based on high-throughput RNA sequencing.Using high-throughput RNA sequencing,a total of 1,233 genes were identified differentially expressed in H5N1 infected A549 cells.DrugBank searches of these 1,233 genes identified 79 validated druggable target genes with 59 approved drugs.We obtained 41 commercially available small-molecule drugs and identified 22 drugs reduced H5N1-mediated A549 cell death.Fifteen of 22 drugs that could significantly recover cell viabilities when administered both pre-and post-H5N1 infection were tested in an H5N1-induced ALI mouse model.Flavin adenine dinucleotide(FAD;an ophthalmic agent for vitamin B2 deficiency),Amitriptyline HCl(an antidepressant drug),calcitriol(an active form of vitamin D)and azacitidine(an anti-neoplastic drug)significantly alleviated H5N1-induced ALI.All four agents markedly decreased the lung injury score and the infiltrating cell count in infected mouse,significantly ameliorated mouse lung edema and significantly improved the survival of H5N1 virus-infected mice.Our study not only identifies novel potential therapies for influenza H5N1 virus-induced lung injury but also provides highly effective and economical screening methods for repurposing drugs that may be generalizable for the prevention and therapy of other diseases.A recent outbreak of pneumonia caused by 2019 novel coronavirus(2019-nCoV,also named SARS-CoV-2)initiated in Wuhan,China,and had been spread to over 100 countries by March 11th of 2020,when the World Health Organization declared it a global pandemic.To further understand the pathogenesis of SARS-CoV-2 caused disease(COVID-19),we measured levels of 48 cytokines and angiotensin ? in the plasmas of 12 COVID-19 patients in Shenzhen,China.The results showed 38 out of 48 cytokines were significantly increased in the plasmas of COVID-19 patients compared to healthy individuals.Seventeen cytokines were associated with SARS-CoV-2 loads.Fifteen cytokines,IL-1?,IL-2,IL-4,IL-7,IL-10,IL-12,IL-17,IP-10,HGF,IL-1ra,M-CSF,G-CSF,IFN-?2,IFN-y,and PDGF-BB were strongly linked to lung injury Murray score,and could predict disease severity of COVID-19 patients with the area under the curve(AUC)of the receiver-operating characteristics(ROC)all above 0.8.Moreover,the Angiotensin ? level in the plasma samples from COVID-19 patients was significantly elevated compared to healthy individuals and linearly linked to viral load and lung injury.Our results indicate that SARS-CoV-2 infection can trigger changes in a wide range of cytokines and angiotensin ?,some of which may be potential biomarkers for the severity of SARS-CoV-2 infection.These findings contribute to improve our understanding of the pathogenesis of this emerging disease.Our results also suggest that modulators of cytokine responses and angiotensin receptor blocker(ARB)drugs may play a therapeutic role in SARS-CoV-2 infection.