Preservation of myocardial function after adenoviral gene transfer in isolated myocardium.
Adenoviral gene transfer to the heart represents a promising model for structure-function analyses. Rabbit hearts were subjected to an ex vivo perfusion protocol that achieves gene transfer in >90% of cardiac myocytes. Contractile function of isolated myocardial preparations of these hearts was then observed for 2 days in a recently developed trabecula culture system. In sham-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, F(init) (14.3 +/- 1. 8 mN/mm(2); n = 21) did not significantly differ from the control (P = 0.75) 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 expression of the reporter genes LacZ coding for beta-galactosidase and Luc coding for firefly luciferase. Luciferase activity increased more than 2,500-fold from background levels of 8. 7 x 10(3 )+/- 5.0 x 10(3) relative light units (RLU)/mg protein (from hearts transfected with promotorless adenovirus with luciferase transgene construct AdNULLLuc, n = 5) to 23.4 x 10(6)+/- 11.1 x 10(6)RLU/mg protein (from hearts tranfected with adenovirus with Rous 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 recombinant adenoviral genes in contracting myocardium without adverse functional effects.
Lehnart
S E
SE
Abteilung Kardiologie und Pneumologie, Universität Göttingen, D-37075 Göttingen, Germany. slehnart@med.uni-goettingen.de
Janssen
P M
PM
Franz
W M
WM
Donahue
J K
JK
Lawrence
J H
JH
Marbán
E
E
Prestle
J
J
Hasenfuss
G
G
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D000818:Animals; D005260:Female; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics / Q000494:pharmacology; D006321:Heart / Q000187:drug effects / Q000502:physiology / Q000821:virology; D007763:Lac Operon / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D009200:Myocardial Contraction / Q000187:drug effects / Q000235:genetics*; D009206:Myocardium / Q000378:metabolism*; D009924:Organ Culture Techniques; D011817:Rabbits; D011943:Receptors, Adrenergic, beta / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D019076:Transgenes / Q000235:genetics
D011943:Receptors, Adrenergic, beta; D008156:Luciferases