Cardiac gene transfer by intracoronary infusion of adenovirus vector-mediated reporter gene in the transplanted mouse heart

This study introduces a model for intracoronary gene transfer in murine cardiac isografts using adenovirus vectors, This approach may offer an opportunity to modulate alloreactivity after cardiac transplantation. Donor hearts were infected via the coronary arteries with a volume of 10(9) plaque-forming units per milliliter of a recombinant adenovirus containing the P-galactosidase-encoding gene (Ad.CMVLacZ). In a control group, 200 mu l of normal saline solution was infused, The grafts were stored in 4 degrees C cold saline solution for 15 minutes, then transplanted heterotopically into syngeneic hosts (B10.BR). The grafts were harvested at 3, 7, 15, or 30 days (n = 5 for each group) after transplantation, and beta-galactosidase activity was assessed by histochemical staining (X-gal), All grafts were functioning when harvested, X-gal staining pattern was nonuniform with positive staining appearing in epicardial, myocardial, and endocardial cells, as well as in the vessel walls, The cells permissive to infection consisted predominantly of myocardial cells, The mean total numbers of beta-gal-positive staining cells per slice were 68.7 +/- 27.3 in the 3 day group, 330.4 +/- 53.8 in the 7-day group, 151.3 +/- 48.0 in the 15-day group, and 39.9 +/- 10.8 in the 30-day group, thus peaking in the 7-day group (p < 0.05). Control isografts (n = 5), retrieved at day 30, revealed no staining activity, In conclusion, our model demonstrates that intracoronary gene transfer to the transplanted murine cardiac grafts is feasible at the time of harvest, Adenovirus-mediated gene transfer produces widespread gene expression which, though perhaps transient, does not adversely affect myocardial structure or function, This technology may allow modification of graft immunogenicity in the future through the production of therapeutic proteins sufficient to modulate local immune responses.