Double-Detargeted Oncolytic Adenovirus Shows Replication Arrest in Liver Cells and Retains Neuroendocrine Cell Killing Ability 英文参考文献.docVIP

Double-DetargetedOncolyticAdenovirusShows ReplicationArrestinLiverCellsandRetains NeuroendocrineCellKillingAbility JustynaLeja1,BerithNilsson1,DiYu1,ElisabetGustafson2,Go¨ranAkerstro? ¨m3,KjellOberg¨ 4 ,Valeria Giandomenico4,MagnusEssand1* 1Division of Clinical Immunology, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden, 2Department of Women’s and Children’s Health, University Hospital, Uppsala,Sweden,3DepartmentofSurgicalSciences,UniversityHospital,Uppsala,Sweden,4DepartmentofMedicalSciences,UniversityHospital,Uppsala,Sweden Abstract Background: We have previously developed an oncolytic serotype 5 adenovirus (Ad5) with chromogranin-A (CgA) promoter-controlledE1Aexpression,Ad[CgA-E1A],withtheintentiontotreatneuroendocrinetumors,includingcarcinoids. Since carcinoids tend to metastasize to the liver it is important to fully repress viral replication in hepatocytes to avoid adenovirus-related liver toxicity. Herein, we explore miRNA-based regulation of E1A expression as a complementary mechanismtopromoter-basedtranscriptionalcontrol. Methodology/PrincipalFindings:Ad[CgA-E1A-miR122],whereE1Aexpressionisfurthercontrolledbysixtandemrepeats of the target sequence for the liver-specific miR122, was constructed and compared to Ad[CgA-E1A]. We observed E1A suppressionandreplicationarrestofthemiR122-detargetedadenovirusinnormalhepatocytes,whilethetwoviruseskilled carcinoid cells to the same degree. Repeated intravenous injections of Ad[CgA-E1A] induced liver toxicity in mice while Ad[CgA-E1A-miR122] injections did not. Furthermore, a miR122-detargeted adenovirus with the wild-type E1A promoter showed reduced replication in hepatic cells compared to wild-type Ad5 but not to the same extent as the miR122- detargetedadenoviruswiththeneuroendocrine-selectiveCgApromoter. Conclusions/Significance:Acombinationoftranscriptional(promoter)andpost-transcriptional(miRNAtarget)regulation tocontrolvirusreplicationmayallowfortheuseofhigherdosesofadenovirusforefficie