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PMID: 9005997 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

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

The Journal of clinical investigation ·Vol. 99 ·No. 2 ·1997-01-15 ·Pages 288-96

Drazner MH, Peppel KC, Dyer S, Grant AO, Koch WJ, Lefkowitz RJ

Abstract

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.

MeSH Terms
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,metabolism Humans Isoproterenol/pharmacology Male Rabbits Receptors, Adrenergic, beta-2/genetics,metabolism Sarcolemma/enzymology Signal Transduction Transgenes
Chemicals
Adrenergic beta-Agonists Receptors, Adrenergic, beta-2 Cyclic AMP Adenylyl Cyclases Isoproterenol
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Drazner M H
Howard Hughes Medical Institute, Department of Medicine (Cardiology), Duke University Medical Center, Durham, North Carolina 27710, USA.
Peppel K C
Dyer S
Grant A O
Koch W J
Lefkowitz R J
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Article Info
Journal
The Journal of clinical investigation
Abbr.
J Clin Invest
ISSN
0021-9738
DOI
10.1172/JCI119157
Published
1997-01-15
Pages
288-96
Language
English
Region
United States
NLM ID
7802877
PMCID
PMC507796
Subset
IM
Grants
NHLBI NIH HHS · HL-16037 · United States
NHLBI NIH HHS · HL32708-12 · United States
NHLBI NIH HHS · T32HL07101 · United States
External Links
PubMed source DOI: 10.1172/JCI119157 PMC Full Text View journal details
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