Genetic modification in organ transplantation and in vivo luciferase imaging

摘要

The genetic modification for organ transplantation is one of the most promising strategies to regulate allogeneicimmune response. Organ-selective gene transfer has especially benefit to control local immune responses. Based on thecatheter technique, we tested to deliver naked plasmid DNA to target graft organs of rats (liver and limbs) by a rapidinjection (hydrodynamics-based transfection). Recent advances in transplantation have been achieved by visualizationof cellular process and delivered gene expression during the inflammatory process by using non-invasive in vivoimaging. Herein, we examined the fate of genetically modified grafts using a firefly luciferase expression plasmid. Forliver modification before transplantation, 6.25% of body weight PBS containing plasmid DNA was injected into theliver through the inferior vena cava using a catheter, and the liver was subsequently transplanted to the recipient rat. Forlimb modification, the femoral caudal epigastric vein was used. In the rat liver transplantation model, substantialluciferase expression was visualized and sustained for only a few days in the grafted liver. We also addressed stressresponses by this hydrodynamics procedure using reporter plasmids containing cis-acting enhancer binding site such asNF-kappa B, cAMP, or heat shock response element. In contrast to hepatic transduction, this genetic limb targetingachieved long lasting luciferase expression in the muscle for 2 months or more. Thus, our results suggest that thiscatheter-based in vivo transfection technique provides an effective strategy for organ-selective gene modification intransplantation, and the bioluminescent imaging is broadening its potential for evaluation to various preclinical studies.