title: Hepatic function is preserved following liver-directed, adenovirus-mediated gene transfer.
abstract: Inherited and acquired disorders of the liver are attractive targets for gene therapy. Hepatic cells are susceptible targets for shuttle vectors because of a diversity of protein and viral receptors and accessibility of a selective afferent blood supply. Preservation of existing hepatic cell integrity and metabolic function is of paramount importance for successful whole animal gene therapy trials. In this report, we examine hepatic cell function and integrity following adenovirus-mediated reporter gene transfer to the liver in vivo. E1-deleted, replication-defective adenovectors encoding the LacZ gene driven by the human CMV promoter were delivered to the liver by isolated portal perfusion. The gene transfer rate, as determined by specific histochemical staining, approached 30% with recombinant protein detectable by Western blot throughout the course of study. Hepatic cell integrity as assessed by histology and hepatic enzyme profile (serum aspartate aminotransferase, gamma-glutamyl transpeptidase) demonstrated normal cellular architecture and no significant difference between transfected liver and controls. Hepatic synthetic and metabolic function, as determined by albumin levels, prothrombin time, and bilirubin, were similar between the two study groups. This study demonstrates that efficient adenovirus-mediated gene transfer and expression in the rat liver do not compromise hepatic cell metabolism and integrity.
authors.affiliation: Department of Surgery, UCLA School of Medicine 90024-6904, USA.
authors.forename: K E
authors.initials: KE
authors.lastname: Drazan
authors.forename: M E
authors.initials: ME
authors.lastname: Csete
authors.forename: X
authors.initials: X
authors.lastname: Da Shen
authors.forename: D
authors.initials: D
authors.lastname: Bullington
authors.forename: G
authors.initials: G
authors.lastname: Cottle
authors.forename: R W
authors.initials: RW
authors.lastname: Busuttil
authors.forename: A
authors.initials: A
authors.lastname: Shaked
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D000818:Animals; D001219:Aspartate Aminotransferases / Q000032:analysis / Q000097:blood; D001483:Base Sequence; D015153:Blotting, Western; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D008099:Liver / Q000201:enzymology / Q000473:pathology / Q000502:physiology*; D016031:Liver Transplantation; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D012313:RNA / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D051381:Rats; D011917:Rats, Inbred Lew; D001616:beta-Galactosidase / Q000235:genetics; D005723:gamma-Glutamyltransferase / Q000032:analysis / Q000097:blood
chemical_list: D017931:DNA Primers; D012313:RNA; D004247:DNA; D005723:gamma-Glutamyltransferase; D001219:Aspartate Aminotransferases; D001616:beta-Galactosidase