Preservation of myocardial function after adenoviral gene transfer in isolated myocardium

S. E. Lehnart, P. M L Janssen, W. M. Franz, J. K. Donahue, J. H. Lawrence, E. Marbán, J. Prestle, G. Hasenfuss

Research output: Contribution to journalArticle

Abstract

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/mm2; n = 12) did not change significantly after 48 h (17.0 ± 1.9 mN/mm2; P = 0.46). In adenovirus-infected preparations, F(init) (14.3 ± 1.8 mN/mm2; n = 21) did not significantly differ from the control (P = 0.75) and was unchanged after 48 h (15.3 ± 2.5 mN/mm2; P = 0.93). After 2 days of continuous contractions, we observed homogenous and high-level expression of the reporter genes LacZ coding for β-galactosidase and Luc coding for firefly luciferase. Luciferase activity increased more than 2,500-fold from background levels of 8.7 x 103 ± 5.0 x 103 relative light units (RLU)/mg protein (from hearts transfected with promotorless adenovirus with luciferase transgene construct AdNULLLuc, n = 5) to 23.4 x 106 ± 11.1 x 106 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.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume279
Issue number3 48-3
StatePublished - 2000

Fingerprint

Myocardium
Luciferases
Adenoviridae
Genes
Transgenes
Galactosidases
Firefly Luciferases
Light
Rous sarcoma virus
Reporter Genes
Cardiac Myocytes
Proteins
Perfusion
Rabbits

Keywords

  • Adenovirus
  • Rabbit
  • Trabecula

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Cite this

Lehnart, S. E., Janssen, P. M. L., Franz, W. M., Donahue, J. K., Lawrence, J. H., Marbán, E., ... Hasenfuss, G. (2000). Preservation of myocardial function after adenoviral gene transfer in isolated myocardium. American Journal of Physiology - Heart and Circulatory Physiology, 279(3 48-3).

Preservation of myocardial function after adenoviral gene transfer in isolated myocardium. / Lehnart, S. E.; Janssen, P. M L; Franz, W. M.; Donahue, J. K.; Lawrence, J. H.; Marbán, E.; Prestle, J.; Hasenfuss, G.

In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 279, No. 3 48-3, 2000.

Research output: Contribution to journalArticle

Lehnart, SE, Janssen, PML, Franz, WM, Donahue, JK, Lawrence, JH, Marbán, E, Prestle, J & Hasenfuss, G 2000, 'Preservation of myocardial function after adenoviral gene transfer in isolated myocardium', American Journal of Physiology - Heart and Circulatory Physiology, vol. 279, no. 3 48-3.
Lehnart, S. E. ; Janssen, P. M L ; Franz, W. M. ; Donahue, J. K. ; Lawrence, J. H. ; Marbán, E. ; Prestle, J. ; Hasenfuss, G. / Preservation of myocardial function after adenoviral gene transfer in isolated myocardium. In: American Journal of Physiology - Heart and Circulatory Physiology. 2000 ; Vol. 279, No. 3 48-3.
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