The Pathogenic Mechanism Research Of MiRNAs In Immunosuppressed Patients With HCMV Infection After Organ Transplantation

Objective Immunosuppressed patients after organ transplantation with HCMV reactivation may cause serious complications even death.So understanding the mechanism of controlling virus latency and reactivation is particularly important.At present antiviral treatment is mainly by inhibiting viral DNA replication,but these antiviral drugs are prone to have side effects and lead to the emergence of resistant strains.The aim of this study is to find miRNAs between latent and active state that may influence HCMV replication and to provide theoretical basis for clinical treatment of HCMV infection.Methods 1.Retrospective analysis 113 cases of immunosuppressive patients and analysis the relation between HCMV reactivation and its complications.2.Determine HCMV latent and active state of differentially expressed miRNAs profile with NGS(next-generation-sequencing)technology.3.GO(gene Ontology)and KEGG(Kyoto Encyclopedia of Genes and Genomes)analysis differentially expressed miRNAs related signaling pathway.4.q PCR(quantitative polymerase chain reaction)verify differentially expressed miRNAs.5.Mi Randa database was used to predict miR-185-5p target gene,and verified by Western Blot.Using miR-185-5p mimics and inhibitors to study the function of miR-185-5p.Results HCMV active infection was found to be associated with GVHD(graft-versus-host disease),asymptomatic HCMV infection and tissue invasive disease after transplantation.Through data processing and analysis,we screened 28 differentially expressed miRNAs.Moreover,RT-PCR validated of miR-185-5p increased significantly when active HCMV infection,further study found that miR-185-5p can decrease Rictor protein expression and increase autophagy.Futhermore,miR-185-5p mimics can promote HCMV replication,while miR-185-5p inhibitors can reduce HCMV replication.Conclusions HCMV can use miRNAs in the cells to control viral latency and reactivation,miR-185-5p can affect autophagy signaling pathway and HCMV replication.