title: Regulated and prolonged expression of mIFN(alpha) in immunocompetent mice mediated by a helper-dependent adenovirus vector.
abstract: A major goal in gene therapy is to develop efficient gene transfer protocols that allow tissue-specific, long-term and tightly regulated expression of the desired transgene. This objective is becoming more attainable through the co-evolution of gene transfer vectors and regulation systems. The ideal vector should efficiently transduce non-dividing cells with minimal toxicity, thus endowing the system with persistent transgene expression. The helper-dependent adenovirus vectors meet these requirements, as demonstrated in various studies in the literature. The most promising regulation system is the tet-on system, which has low basal transcriptional activity and high inducibility. To explore the regulated transgene expression in the context of a helper-dependent vector, we constructed the HD-TET-IFN vector, containing the mIFN(alpha) gene under the control of the tetracycline inducible transactivator rtTA2(s)-S2. Mice injected with HD-TET-IFN showed high levels of serum mIFN(alpha) only upon transcriptional activation. The transgene expression was reinducible to the same high level up to 3 months p.i., and the amount of expressed cytokine could be regulated by dosing doxycycline. Transcriptional activation of mIFN(alpha) induced by doxycycline resulted in prolonged survival and reduced liver damage in HD-TET-IFN-injected mice challenged with a lethal dose of coronavirus. Activation of antiviral genes mediated by doxycycline-dependent mIFN(alpha) expression was also observed at low HD-TET-IFN doses. The possibility of controlling gene expression by the combination of HD vectors and the latest tet-on transactivator also holds promise for studying gene function in other animal models.
authors.affiliation: IRBM P Angeletti, Rome, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Aurisicchio
authors.forename: H
authors.initials: H
authors.lastname: Bujard
authors.forename: W
authors.initials: W
authors.lastname: Hillen
authors.forename: R
authors.initials: R
authors.lastname: Cortese
authors.forename: G
authors.initials: G
authors.lastname: Ciliberto
authors.forename: N
authors.initials: N
authors.lastname: La Monica
authors.forename: F
authors.initials: F
authors.lastname: Palombo
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D000900:Anti-Bacterial Agents / Q000627:therapeutic use*; D006528:Carcinoma, Hepatocellular; D002460:Cell Line; D004318:Doxycycline / Q000627:therapeutic use*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D015870:Gene Expression; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D006524:Hepatitis, Viral, Animal / Q000628:therapy; D006801:Humans; D016898:Interferon-alpha / Q000032:analysis / Q000097:blood / Q000235:genetics*; D008099:Liver / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D023421:Models, Animal; D012333:RNA, Messenger / Q000032:analysis; D015534:Trans-Activators / Q000235:genetics; D019076:Transgenes
chemical_list: D000900:Anti-Bacterial Agents; D016898:Interferon-alpha; D012333:RNA, Messenger; D015534:Trans-Activators; D000410:Alanine Transaminase; D004318:Doxycycline