A new surgical approach to improve gene transfer in liver using lentiviral vectors.
OBJECTIVE
Metabolic inherited liver diseases are attractive targets for gene therapy. Recombinant lentiviruses are very powerful viral vectors able to infect nonmitotic cells. We wanted to develop a new surgical approach to improve gene transfer in adult liver using low viral doses.


METHODS
Adult rats were injected with 2.108 infectious particles of lentiviral vectors encoding the green fluorescent protein marker gene under control of a liver-specific promoter transthyretin. In the control group (n = 5), gene delivery was performed by inflow intraportal injection. In the surgical group (n = 5), liver was completely excluded from systemic circulation before viral injection in infrahepatic vena cava with high pressure.


RESULTS
At day 9, transduction efficiency was 14.35% in the surgical group 3 and 0.39% in the control group (P = .016). At month 2, the number of transduced hepatocytes decreased in the most part of rats, except in half of rats in the surgical group. Antibodies against green fluorescent protein were detected in all rats at month 2, except one in the surgical group.


CONCLUSIONS
We developed a new surgical approach allowing an efficient transduction of hepatocytes in adult rats using lentivirus at low viral doses. We have now to control the immune response to permit long-term expression of transgene.
Dariel
Anne
A
Pediatric Surgery Department, University Hospital, 44093 Nantes, France.
Nguyen
Tuan Huy
TH
Pichard
Virginie
V
Schmitt
Françoise
F
Aubert
Dominique
D
Ferry
Nicolas
N
Podevin
Guillaume
G
D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D022781:Hepatocytes / Q000378:metabolism; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics*; D008099:Liver / Q000098:blood supply; D008102:Liver Circulation; D008297:Male; D010957:Plasmids; D051381:Rats; D011911:Rats, Gunn; D014161:Transduction, Genetic; D014684:Venae Cavae
D049452:Green Fluorescent Proteins