The long-term objective of this project is to make gene therapy based prevention of coronary and peripheral vascular graft-induced occlusion and restenosis feasible by developing a safe, effective and economical DNA delivery method based on electroporation. Electroporation employs electric field pulses to make cell membranes permeable for DNA, resulting in a 100-1000 fold increase of intracellular DNA delivery as compared to the "naked DNA" method. The proposed project involves designing and testing suitable electrodes and optimizing electroporation parameters in ex vivo organ culture, aimed eventually at applications for vein grafts in clinical settings. The specific aims of this project are: (1) to evaluate the efficiency of pulsed electric fields for ex vivo DNA transfer into cells of rat inferior vena cava and porcine saphenous vein, respectively. (2) to evaluate vasospastic & vasorelaxation effects of pulsed electric fields and (3) to design, develop and test electrodes based on experimental observations and theoretical computational modeling of electric fields. This investigational effort may provide a critical advancement for the delivery of therapeutic genes to the interstitial spaces and to the cyotosol with high efficiency and few side effects at a reasonable cost, superior to methods that have been tried in the past. The commercial product will be an innovative electroporation device and method for the efficient local delivery of therapeutic genes or compounds to enhance the success rate of vascular grafting for the prevention of vessel graft dysfunctions. Reducing the incidence of graft failure will save or prolong patients' lives and prevent the necessity of re-treatments which deplete scarce human as well as financial healthcare resources. The project goal will be to "condition" vein-graft ex vivo via gene transfer prior to its interpositioning with the target artery. In clinical settings the ex vivo treatment approach can be easily incorporated into the usual graft surgical procedure. It only requires the treatment of the graft before implantation.
Thesaurus Terms: blood vessel restoration, disease /disorder prevention /control, electric field, electrode, electroporation, gene delivery system, gene therapy, iatrogenic disease, method development blood vessel occlusion, blood vessel transplantation, coronary vessel, membrane permeability, peripheral blood vessel, restenosis, transplant rejection bioengineering /biomedical engineering, biotechnology, laboratory rat, organ culture, swine
