Pig liver gene therapy by noninvasive interventionist catheterism.
The efficacy of noninvasive interventionist catheterism in large animals as an alternative to the hydrodynamic procedure, described for small animals, is evaluated. Basically, gene transfer is performed by implantation and fixation of a balloon catheter within the suprahepatic vein of anesthetized pigs, through the femoral vein. The catheter tip is identified by fluoroscopy, injecting a contrast solution that marks large or small hepatic territories. Animals were injected with a 100 ml pTG7101 plasmid solution (40 microg/ml), which contains the human alpha-1 antitrypsin gene, perfused at a rate of 7.5 ml/s and efficacy and toxicity of the procedure were evaluated. The results show: (i) the highest efficacy in protein production is reached when perfusion is limited to small areas of the liver; (ii) no relevant hepatic toxicity was observed; (iii) gene transfer is mainly located in the areas around the central vein, as seen in the immunohistochemical studies; (iv) the electron microscopy studies indicate that the areas with good transfection efficacy show the presence of abundant endocytic vesicles that may even fuse among themselves. These data suggest that retrovenous injection by noninvasive interventionist catheterism could become an efficient procedure for hepatic gene transfer with potential clinical applications.
Aliño
S F
SF
Departamento de Farmacología, Facultad de Medicina, Universidad de Valencia, Valencia, Spain. alino@uv.es
Herrero
M J
MJ
Noguera
I
I
Dasí
F
F
Sánchez
M
M
D000818:Animals; D002404:Catheterization; D004247:DNA / Q000008:administration & dosage*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D007150:Immunohistochemistry; D008099:Liver / Q000378:metabolism*; D008107:Liver Diseases / Q000378:metabolism / Q000628:therapy; D046529:Microscopy, Electron, Transmission; D023421:Models, Animal; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D014162:Transfection / Q000379:methods*; D000515:alpha 1-Antitrypsin / Q000032:analysis / Q000235:genetics* / Q000378:metabolism
D000515:alpha 1-Antitrypsin; D004247:DNA