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- 저자 : ( Zongyi Hu ) (검색결과 2 건)
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( Hyung Joon Yim ),( Billy Lin ),( Shanshan He ),( Zongyi Hu ),( T. Jake Liang ) 대한간학회 2016 춘·추계 학술대회 (KASL) Vol.2016 No.1
Aims: Several directly acting antiviral agents (DAA) were currently approvedfor the treatment of chronic hepatitis C (CHC). AlthoughDAA therapies are associated with better tolerability and improvedresponse rates, occurrence of drug resistance has been the drawback.The aim of the present study is to develop full-length resistance associatedvariants (RAV) HCV culture systems to evaluate the efficacyand the cross resistance of current antiviral drugs for future therapeuticstrategies.Methods: Resistance associated substitutions on NS3 (A156T, D168V),NS5A (L31V, Y93H, L31V+Y93H), and NS5B (S282T) domains weregenerated by site directed mutagenesis and cloned into genotype2a J6/JFH1 HCV plasmid with or without luciferase gene. After In vitro RNA transcription, RNAs of RAV were transfected into Huh 7.5.1cells. HCVcc in the supernatants were collected and used for thereinfection and treatment experiment to confirm drug susceptibility.We performed HCV core staining and Renilla luciferease assays toassess treatment response to multiple DAAs and other antiviral drugswith different mechanism of action after transfection or infectionof RAVs.Results: DAAs in the same classes shared cross resistance to correspondingRAVs: boceprevir, telaprevir, simeprevir, and asunaprevirto NS3 RAVs; daclatasvir and ledipasvir to NS5A RAVs; sofosbuvirto NS5B RAV. However, DAAs of the other classes effectively suppressedRAVs as well as wild type. All the RAVs were sensitive todrugs with different action of mechanism including interferon alfa,ribavirin, cyclosporine (cyclophilin inhibitor) or chlorcyclizine (entry inhibitors).Conclusions: We developed full-length RAV HCV culture systemsbased on genotype 2a strain. This system will be useful to assessantiviral response of drugs with different action of mechanisms.Combination of different classes of DAA or new drugs with differentaction mechanisms (e.g. cyclophilin inhibitor or entry inhibitors) shouldbe a future therapeutic strategy for overcoming drug resistance inthe treatment of CHC.
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