title: Sustained inhibition of hepatitis B virus replication in vivo using RNAi-activating lentiviruses.
abstract: Chronic infection with hepatitis B virus (HBV) puts individuals at high risk for complicating cirrhosis and liver cancer, but available treatment to counter the virus rarely eliminates infection. Although harnessing RNA interference (RNAi) to silence HBV genes has shown the potential, achieving efficient and durable silencing of viral genes remains an important goal. Here we report on the propagation of lentiviral vectors (LVs) that successfully deliver HBV-targeting RNAi activators to liver cells. Mono- and tricistronic artificial primary microRNAs (pri-miRs) derived from pri-miR-31, placed under transcriptional control of the liver-specific modified murine transthyretin (mTTR) promoter, caused efficient inhibition of HBV replication markers. The tricistronic cassette was capable of silencing a mutant viral target and the effects were observed without disrupting the function of an endogenous miR (miR-16). The mTTR promoter stably expressed a reporter transgene in mouse livers over a study period of 1 year. Good silencing of HBV genes, without evidence of toxicity, was demonstrated following intravenous injection of LVs into neonatal HBV transgenic mice. Collectively, these data indicate that LVs may achieve sustained inhibition of HBV replication that is appealing for their therapeutic use.
authors.affiliation: Antiviral Gene Therapy Research Unit, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, Gauteng, South Africa.
authors.forename: D
authors.initials: D
authors.lastname: Ivacik
authors.affiliation: Antiviral Gene Therapy Research Unit, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, Gauteng, South Africa.
authors.forename: A
authors.initials: A
authors.lastname: Ely
authors.affiliation: Agence Nationale de Sécurité du Médicament et des produits de Santé, Saint Denis, France.
authors.forename: N
authors.initials: N
authors.lastname: Ferry
authors.affiliation: Antiviral Gene Therapy Research Unit, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, Gauteng, South Africa.
authors.forename: P
authors.initials: P
authors.lastname: Arbuthnot
mesh_terms: D000818:Animals; D001483:Base Sequence; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D057809:HEK293 Cells; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000502:physiology*; D019694:Hepatitis B, Chronic / Q000628:therapy* / Q000821:virology; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008099:Liver / Q000378:metabolism / Q000821:virology; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000235:genetics; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011228:Prealbumin / Q000235:genetics; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D019076:Transgenes; D054334:Viral Regulatory and Accessory Proteins; D014779:Virus Replication
chemical_list: D035683:MicroRNAs; C522604:Mirn16 microRNA, mouse; D011228:Prealbumin; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein; D049409:Luciferases, Firefly