Enhanced plasma cholesterol lowering effect of retrovirus-mediated LDL receptor gene transfer to WHHL rabbit liver after improved surgical technique and stimulation of hepatocyte proliferation by combined partial liver resection and thymidine kinase--ganciclovir treatment.
In this study we report an improved method for in vivo gene transfer to liver. Repeated injections of Moloney murine leukemia virus-derived retroviruses containing LDL receptor cDNA were given to the portal vein in combination with a 10% partial liver resection and stimulation of hepatocyte proliferation by plasmid/liposome-mediated thymidine kinase gene transfer and ganciclovir treatment. The method was used for the treatment of LDL receptor deficiency in Watanabe heritable hyperlipidemic rabbits. We demonstrate an increase in hepatocyte proliferation index by thymidine kinase and ganciclovir treatment from 0.9 to 1.35% and a maximum of 35% decrease in total plasma cholesterol level 2-3 months after the gene transfer. A 20% decline was still present after a 52-week follow-up period. A 50% decrease was also observed in plasma triglycerides. Liver function tests indicated a transient increase in plasma alkaline phosphatase level up to 12 weeks after the gene transfer. In situ PCR and RT-PCR analyses indicated that the transgene was present in periportal areas and was transcribed to mRNA 1 week after the gene transfer. Because of the relatively simple and controllable technique we suggest that repeated retrovirus injections via a portal vein catheter together with the limited partial liver resection and plasmid/liposome-mediated thymidine kinase gene transfer-ganciclovir treatment may be used to improve the results of retrovirus-mediated liver gene therapy.
Pakkanen
T M
TM
AI Virtanen Institute, University of Kuopio, Finland.
Laitinen
M
M
Hippeläinen
M
M
Kallionpää
H
H
Lehtolainen
P
P
Leppänen
P
P
Luoma
J S
JS
Tarvainen
R
R
Alhava
E
E
Ylä-Herttuala
S
S
D000818:Animals; D000963:Antimetabolites / Q000627:therapeutic use; D002455:Cell Division / Q000187:drug effects; D002784:Cholesterol / Q000097:blood*; D005260:Female; D015774:Ganciclovir / Q000627:therapeutic use; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006938:Hyperlipoproteinemia Type II / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D008099:Liver / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D008297:Male; D011817:Rabbits; D011973:Receptors, LDL / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014280:Triglycerides / Q000097:blood
D000963:Antimetabolites; D011973:Receptors, LDL; D014280:Triglycerides; D002784:Cholesterol; D013937:Thymidine Kinase; D015774:Ganciclovir