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J Biol Chem, Vol. 273, Issue 51, 34263-34271, December 18, 1998

Adenovirus-mediated Gene Transfer of cGMP-dependent Protein Kinase Increases the Sensitivity of Cultured Vascular Smooth Muscle Cells to the Antiproliferative and Pro-apoptotic Effects of Nitric Oxide/cGMP

Jean-Daniel Chicheabc, Stacy M. Schlutsmeyerb, Donald B. Bloche, Suzanne M. de la Montef, Jesse D. Roberts Jr.ab, Galina Filippovb, Stefan P. Janssensg, Anthony Rosenzweigbh, and Kenneth D. Blochbhi

From the a Department of Anesthesia and Critical Care, the b Cardiovascular Research Center, the h Cardiology Division, the e Arthritis Unit, and the f Molecular Hepatology Division of the General Medical Services, Massachusetts General Hospital and the Departments of c Anesthesia and i Medicine, Harvard Medical School, Charlestown, Massachusetts 02129 and the g Center for Transgene Technology and Gene Therapy, University of Leuven, B-3000 Leuven, Belgium

Studies in vitro have underestimated the importance of cGMP-dependent protein kinase (PKG) in the modulation of vascular smooth muscle cell (SMC) proliferation and apoptosis in vivo. This is attributable, in part, to a rapid decline in PKG levels as vascular SMC are passaged in culture. We used a recombinant adenovirus encoding PKG (Ad.PKG) to augment kinase activity in cultured rat pulmonary artery SMC (RPaSMC). Incubation of Ad.PKG-infected RPaSMC (multiplicity of infection = 200) with 8-Br-cGMP decreased serum-stimulated DNA synthesis by 85% and cell proliferation at day 5 by 74%. The effect of 8-Br-cGMP on DNA synthesis in Ad.PKG-infected RPaSMC was blocked by KT5823 (PKG inhibitor), but not by KT5720 (cAMP-dependent protein kinase inhibitor). A nitric oxide (NO) donor compound, S-nitrosoglutathione, at concentrations as low as 100 nM, inhibited DNA synthesis in Ad.PKG-infected RPaSMC, but not in uninfected cells or in cells infected with a control adenovirus. In addition, 8-Br-cGMP and S-nitrosoglutathione induced apoptosis in serum-deprived RPaSMC infected with Ad.PKG, but not in uninfected cells or in cells infected with a control adenovirus. These results demonstrate that modulation of PKG levels in vascular SMC can alter the sensitivity of these cells to NO and cGMP. Moreover, these observations suggest an important role for PKG in the regulation of vascular SMC proliferation and apoptosis by NO and cGMP.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.


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