Potentiation of beta-adrenergic signaling by adenoviral-mediated gene transfer in adult rabbit ventricular myocytes

Our laboratory has been testing the hypothesis that genetic modulation of the beta-adrenergic signaling cascade can enhance cardiac function. We have previously shown that transgenic mice with cardiac overexpression of either the human beta2-adrenergic receptor (beta2AR) or an inhibitor of the beta-...

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Bibliographic Details
Published in:The Journal of clinical investigation 1997-01, Vol.99 (2), p.288-296
Main Authors: Drazner, M H, Peppel, K C, Dyer, S, Grant, A O, Koch, W J, Lefkowitz, R J
Format: Article
Language:English
Subjects:
Adenoviridae - genetics
Adenylyl Cyclases - analysis
Adrenergic beta-Agonists - pharmacology
Animals
Cell Survival
Cells, Cultured
Cyclic AMP - metabolism
Gene Transfer Techniques
Genetic Vectors
Heart Ventricles - cytology
Heart Ventricles - metabolism
Humans
Isoproterenol - pharmacology
Male
Rabbits
Receptors, Adrenergic, beta-2 - genetics
Receptors, Adrenergic, beta-2 - metabolism
Sarcolemma - enzymology
Signal Transduction
Transgenes
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Summary:Our laboratory has been testing the hypothesis that genetic modulation of the beta-adrenergic signaling cascade can enhance cardiac function. We have previously shown that transgenic mice with cardiac overexpression of either the human beta2-adrenergic receptor (beta2AR) or an inhibitor of the beta-adrenergic receptor kinase (betaARK), an enzyme that phosphorylates and uncouples agonist-bound receptors, have increased myocardial inotropy. We now have created recombinant adenoviruses encoding either the beta2AR (Adeno-beta2AR) or a peptide betaARK inhibitor (consisting of the carboxyl terminus of betaARK1, Adeno-betaARKct) and tested their ability to potentiate beta-adrenergic signaling in cultured adult rabbit ventricular myocytes. As assessed by radioligand binding, Adeno-beta2AR infection led to approximately 20-fold overexpression of beta-adrenergic receptors. Protein immunoblots demonstrated the presence of the Adeno-betaARKct transgene. Both transgenes significantly increased isoproterenol-stimulated cAMP as compared to myocytes infected with an adenovirus encoding beta-galactosidase (Adeno-betaGal) but did not affect the sarcolemmal adenylyl cyclase response to Forskolin or NaF. beta-Adrenergic agonist-induced desensitization was significantly inhibited in Adeno-betaARKct-infected myocytes (16+/-2%) as compared to Adeno-betaGal-infected myocytes (37+/-1%, P < 0.001). We conclude that recombinant adenoviral gene transfer of the beta2AR or an inhibitor of betaARK-mediated desensitization can potentiate beta-adrenergic signaling.
ISSN:0021-9738
DOI:10.1172/JCI119157