Selective gene transfer into the liver of non-human primates with E1-deleted, E2A-defective, or E1-E4 deleted recombinant adenoviruses.
Preclinical studies were designed to investigate the feasibility and safety of recombinant adenoviruses transduced into the hepatic artery of nonhuman primates. The vectors used are recombinant adenoviruses deleted in E1 and contain either a temperature-sensitive mutation in the E2a gene, which encodes a defective DNA-binding protein at nonpermissive temperatures, or a deletion of the E4 region, including open reading frame (ORF) 6. Six 8- to 10-kg baboons underwent femoral artery cannulation, and angiographic techniques were used to introduce vector selectively into either a portion of the right lobe of the liver via a branch of the right hepatic artery or the common hepatic artery. Necropsies were performed at 4, 29, or 61 days. Serial sequential liver biopsies were performed in the baboons that survived 29 or 61 days. In the 2 baboons with vector transduction into the right hepatic artery, X-Gal histochemical analysis of the liver showed evidence of quantitatively increased gene transfer in the targeted lobe; however, gene transfer was present throughout the liver. Quantitative analysis of histopathology showed that portal inflammation was present throughout both livers transduced with the highest dose of vector. No differences were seen in the level of portal inflammation in targeted and untargeted lobes despite the observed qualitative and quantitative differences in gene expression. Southern blot analysis of total cellular DNA isolated from targeted and nontargeted lobes showed similar levels of viral DNA throughout the liver. Polymerase chain reaction (PCR) analysis was able to detect viral DNA sequence in gonads and brain as well as many other tissues in baboons treated with high-dose vector. In baboons treated with lower doses of an E1-E4 deleted vector expressing the human ornithine transcarbamylase (OTC) gene, DNA was detectable by nested PCR in liver but not gonads at days 29 and 61. The data suggest that intraarterial administration of recombinant adenoviral E1-E4 deleted vector is feasible and safe. At high doses of vector, widespread dissemination of vector DNA is seen. At low doses, hepatic gene transfer is not associated with vector DNA dissemination to gonads.
Raper
S E
SE
Institute for Human Gene Therapy, Harrison Department of Surgical Research, University of Pennsylvania School of Medicine, Philadelphia 19104-6100, USA.
Haskal
Z J
ZJ
Ye
X
X
Pugh
C
C
Furth
E E
EE
Gao
G P
GP
Wilson
J M
JM
D000256:Adenoviridae / Q000235:genetics*; D017498:Adenovirus E1 Proteins / Q000235:genetics; D017502:Adenovirus E2 Proteins / Q000235:genetics; D017504:Adenovirus E4 Proteins / Q000235:genetics; D000818:Animals; D004279:DNA, Viral; D003673:Defective Viruses / Q000235:genetics; D005240:Feasibility Studies; D017353:Gene Deletion; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage; D006066:Gonads / Q000378:metabolism; D008099:Liver / Q000737:chemistry / Q000378:metabolism* / Q000473:pathology; D009954:Ornithine Carbamoyltransferase / Q000235:genetics; D010215:Papio
D017498:Adenovirus E1 Proteins; D017502:Adenovirus E2 Proteins; D017504:Adenovirus E4 Proteins; D004279:DNA, Viral; D009954:Ornithine Carbamoyltransferase