Preservation of myocardial function after adenoviral gene transfer in isolated myocardium

Adenoviral gene transfer to the heart represents a promising model for stru cture-function analyses. Rabbit hearts were subjected to an ex vivo perfusi on protocol that achieves gene transfer in >90% of cardiac myocytes. Contra ctile function of isolated myocardial preparations of these hearts was then observed for 2 days in a recently developed trabecula culture system. In s ham-infected hearts, the initial developed force (F-init) (15.6 +/- 3.7 mN/ mm(2); n = 12) did not change significantly after 48 h (17.0 +/- 1.9 mN/mm( 2); P = 0.46). In adenovirus-infected preparations, Finit (14.3 coproduct 1 .8 mN/mm(2); n = 21) did not significantly differ from the control (P = 0.7 5) and was unchanged after 48 h (15.3 +/- 2.5 mN/mm(2); P = 0.93). After 2 days of continuous contractions, we observed homogenous and high-level expr ession of the reporter genes LacZ coding for beta-galactosidase and Luc cod ing for firefly luciferase. Luciferase activity increased more than 2,500-f old from background levels of 8.7 x 10(3) +/- 5.0 x 10(3) relative light un its (RLU)/mg protein (from hearts transfected with promotorless adenovirus with luciferase transgene construct AdNULLLuc, n = 5) to 23.4 x 10(6) +/- 1 1.1 x 10(6) RLU/mg protein (from hearts tranfected with adenovirus with Rou s sarcoma virus promotor and luciferase transgene construct AdRSVLuc, n = 5 ) in infected myocardial preparations (P < 0.005). Our results demonstrate a new ex vivo approach to achieve homogenous and high-level expression of r ecombinant adenoviral genes in contracting myocardium without adverse funct ional effects.