Human AAT gene transfer to pig liver improved by using a perfusion isolated organ endovascular procedure.
OBJECTIVE
The efficiency of endovascular liver gene transfer in pigs is evaluated by comparing two models of retrograde catheterization: single lobe catheterization with portal inflow (open procedure) versus whole liver isolation with portal and inferior vena cava blockage (close procedure).


METHODS
Percutaneous endovascular catheterization was performed in pigs. Open procedure (n = 3): 8Fr balloon catheter placement in a suprahepatic branch through the jugular vein. Closed procedure (n = 3): simultaneous catheterization of the intrahepatic portal vein (transhepatic catheterization, 10Fr balloon catheter), the supra- and infrahepatic cava veins (8Fr balloon catheters through the jugular and femoral veins). In both models, 200 ml of hAAT DNA solution (20 μg/ml) were retrogradely injected at 20 ml/s. Tissue samples (8 per liver) were obtained 14 days later and the exogenous DNA, RNA and protein per cell were quantified. Blood samples were collected periodically for transaminase determination in all the animals.


RESULTS
The open procedure achieved lower (approx. 1000-fold) DNA delivery, resulting in a significantly lower (p < 0.001) gene transcription (> 100-fold). The closed model also achieved a higher translation index, although differences were smaller (p < 0.001).


CONCLUSIONS
Portal inflow blockage increased the delivery, transcription and translation indexes, significantly improving the final procedure efficacy when compared with an open procedure.


CONCLUSIONS
Endovascular hydrodynamic pig liver gene transfer: open procedure versus closed procedure. Open procedure resulted in much lower DNA delivery than closed model. Open procedure reached significantly lower gene transcription index. Translation index with closed model was higher than with the open.
Departamento de Farmacología, Facultad de Medicina, Universidad de Valencia, Valencia, Spain.
Luis
L
Sendra
Servicio de Radiología, Hospital Universitario y Politécnico La Fe, Grupo de Investigación Biomédica de Imagen IIS La Fe, Valencia, Spain.
Daniel
D
Pérez
Departamento de Farmacología, Facultad de Medicina, Universidad de Valencia, Valencia, Spain.
Antonio
A
Miguel
Departamento de Farmacología, Facultad de Medicina, Universidad de Valencia, Valencia, Spain. maria.jose.herrero@uv.es.|Unidad de Farmacogenética, IIS La Fe, Servicio de Farmacia, Hospital Universitario y Politécnico La Fe, Valencia, Spain. maria.jose.herrero@uv.es.
María José
MJ
Herrero
SCSIE, Servicio Central de Soporte a la Investigación Experimental, Universidad de Valencia, Valencia, Spain.
Inmaculada
I
Noguera
SCSIE, Servicio Central de Soporte a la Investigación Experimental, Universidad de Valencia, Valencia, Spain.
Ana
A
Díaz
Instituto de Biomedicina de Valencia, CSIC, Valencia, Spain.
Domingo
D
Barettino
Servicio de Radiología, Hospital Universitario y Politécnico La Fe, Grupo de Investigación Biomédica de Imagen IIS La Fe, Valencia, Spain.
Luis
L
Martí-Bonmatí
Departamento de Farmacología, Facultad de Medicina, Universidad de Valencia, Valencia, Spain.|Unidad de Farmacogenética, IIS La Fe, Servicio de Farmacia, Hospital Universitario y Politécnico La Fe, Valencia, Spain.|Unidad de Farmacología Clínica, ACM Hospital Universitario y Politécnico La Fe, Valencia, Spain.
Salvador F
SF
Aliño
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D057510:Endovascular Procedures / Q000379:methods*; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D008099:Liver; D008107:Liver Diseases / Q000628:therapy*; D010477:Perfusion / Q000379:methods*; D013552:Swine
C470011:CFAP91 protein, human; D002352:Carrier Proteins; D004247:DNA
Catheter; Hydrodynamic; Naked DNA; Retrograde injection; Vascular exclusion