hari sundar

A translatable cardiac surgical procedure is developed that allows for complete surgical isolation of the heart in situ with retrograde (via the coronary venous circulation) administration of both vector and endothelial permeabilizing agents to increase myocyte transduction efficiency. In six adult dogs the heart was completely isolated with tourniquets placed around both vena cavae and cannulae and all pulmonary veins. On CPB, the aorta and pulmonary artery were cross-clamped and the heart isolated. Crystalloid cardioplegia at 4°C containing 1013 particles of adenovirus encoding LacZ and 15 g VEGF was infused retrograde into the coronary sinus and recirculated for a total of 30 minutes. The dogs were then weaned from CPB and allowed to recover. Using a catheter, three control dogs underwent retrograde infusion of the same cocktail without cardiac isolation or CPB. �galactosidase activities in the CPB group were several orders of magnitude higher in both the right and left ventricles when compared to the control group (p < 0.05). X-gal staining from the CPB group showed unequivocal evidence of myocyte gene expression globally in a significant proportion of cardiac myocytes. No myocyte gene expression was observed in the control group. This approach with cardiac isolation and retrograde delivery of vector via the coronary sinus results in efficient myocyte transduction in an adult large animal in vivo.