Portal branch ligation induces efficient retrovirus-mediated gene delivery in rat liver.
BACKGROUND
The in vivo transduction of hepatocytes with conventional retrovirus vectors requires the induction of cell division and this can currently only be achieved by invasive surgery or by inducing severe liver damage. We hypothesised that partial portal branch ligation (PBL) could induce hepatocyte proliferation and efficient gene transfer in the rat.


METHODS
We ligated the portal branch serving 70% of the liver and measured the kinetics of liver mass restoration and cell proliferation and the distribution of dividing hepatocytes after administration of 5-bromo-2'-deoxyuridine. The efficiency of retrovirus-mediated gene transfer after PBL was tested by use of beta-galactosidase-expressing recombinant retroviruses. The viruses were administered in a single injection via the portal vein at different times after PBL and the livers of transduced animals were analysed 4 days later.


RESULTS
We found that the number of cycling hepatocytes remained stable between 24 and 44 h after PBL (approximately 12.5%). Although there was a high level of inter-animal variability, hepatocyte proliferation was always initiated in the same lobe of the liver. In animals that had undergone PBL, 19% of hepatocytes were transduced 28 h after the administration of a single high-titre injection of retroviruses, mainly around the portal spaces.


CONCLUSIONS
PBL can mediate the efficient transduction of hepatocytes in vivo after a single intravenous injection of recombinant retroviruses. This approach is feasible in humans.
Lorand
Isabelle
I
INSERM EMI 00-20 and University Paris XI, Hôpital Antoine-Béclère, 92141 Clamart, France.
Vons
Corinne
C
Nguyen
Tuan
T
Di Rico
Virginie
V
Coulomb
Aurore
A
Franco
Dominique
D
Capron
Frédérique
F
Weber
Anne
A
D000818:Animals; D049109:Cell Proliferation; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D008026:Ligation; D008099:Liver / Q000378:metabolism* / Q000502:physiology; D008115:Liver Regeneration; D008297:Male; D011169:Portal Vein; D051381:Rats; D017208:Rats, Wistar; D012190:Retroviridae / Q000235:genetics*