Phosphodiesterase-5A (PDE5A) is localized to the endothelial caveolae and modulates NOS3 activity

Milena A. Gebska, Blake K. Stevenson, Anna R. Hemnes, Trinity Bivalacqua, Azeb Haile, Geoffrey G. Hesketh, Christopher I. Murray, Ari L. Zaiman, Marc K Halushka, Nispa Krongkaew, Travis D. Strong, Carol A. Cooke, Hazim El-Haddad, Rubin M. Tuder, Dan E Berkowitz, Hunter C. Champion

Research output: Contribution to journalArticle

Abstract

AimsIt has been well demonstrated that phosphodiesterase-5A (PDE5A) is expressed in smooth muscle cells and plays an important role in regulation of vascular tone. The role of endothelial PDE5A, however, has not been yet characterized. The present study was undertaken to determine the presence, localization, and potential physiologic significance of PDE5A within vascular endothelial cells.Methods and resultsWe demonstrate primary location of human, mouse, and bovine endothelial PDE5A at or near caveolae. We found that the spatial localization of PDE5A at the level of caveolin-rich lipid rafts allows for a feedback loop between endothelial PDE5A and nitric oxide synthase (NOS3). Treatment of human endothelium with PDE5A inhibitors resulted in a significant increase in NOS3 activity, whereas overexpression of PDE5A using an adenoviral vector, both in vivo and in cell culture, resulted in decreased NOS3 activity and endothelium-dependent vasodilation. The molecular mechanism responsible for these interactions is primarily regulated by cGMP-dependent second messenger. PDE5A overexpression also resulted in a significant decrease in protein kinase 1 (PKG1) activity. Overexpression of PKG1 rapidly activated NOS3, whereas silencing of the PKG1 gene with siRNA inhibited both NOS3 phosphorylation (S1179) and activity, indicating a novel role for PKG1 in direct regulation of NOS3.ConclusionOur data collectively suggest another target for PDE5A inhibition in endothelial dysfunction and provide another physiologic significance for PDE5A in the modulation of endothelial-dependent flow-mediated vasodilation. Using both in vitro and in vivo models, as well as human data, we show that inhibition of endothelial PDE5A improves endothelial function.

Original languageEnglish (US)
Pages (from-to)353-363
Number of pages11
JournalCardiovascular Research
Volume90
Issue number2
DOIs
StatePublished - May 1 2011

Fingerprint

Type 5 Cyclic Nucleotide Phosphodiesterases
Caveolae
Vasodilation
Endothelium
Caveolins
Phosphodiesterase Inhibitors
Second Messenger Systems
Gene Silencing
Nitric Oxide Synthase
Protein Kinases
Small Interfering RNA
Smooth Muscle Myocytes
Blood Vessels

Keywords

  • Adenoviral transfection
  • Caveolin-1
  • cGMP
  • Endothelial cells
  • Lipid rafts
  • NOS3
  • PDE5A
  • Phosphodiesterase
  • Primary human endothelial cells
  • Pulmonary circulation
  • Signalling pathway
  • Sildenafil

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)
  • Physiology

Cite this

Gebska, M. A., Stevenson, B. K., Hemnes, A. R., Bivalacqua, T., Haile, A., Hesketh, G. G., ... Champion, H. C. (2011). Phosphodiesterase-5A (PDE5A) is localized to the endothelial caveolae and modulates NOS3 activity. Cardiovascular Research, 90(2), 353-363. https://doi.org/10.1093/cvr/cvq410

Phosphodiesterase-5A (PDE5A) is localized to the endothelial caveolae and modulates NOS3 activity. / Gebska, Milena A.; Stevenson, Blake K.; Hemnes, Anna R.; Bivalacqua, Trinity; Haile, Azeb; Hesketh, Geoffrey G.; Murray, Christopher I.; Zaiman, Ari L.; Halushka, Marc K; Krongkaew, Nispa; Strong, Travis D.; Cooke, Carol A.; El-Haddad, Hazim; Tuder, Rubin M.; Berkowitz, Dan E; Champion, Hunter C.

In: Cardiovascular Research, Vol. 90, No. 2, 01.05.2011, p. 353-363.

Research output: Contribution to journalArticle

Gebska, MA, Stevenson, BK, Hemnes, AR, Bivalacqua, T, Haile, A, Hesketh, GG, Murray, CI, Zaiman, AL, Halushka, MK, Krongkaew, N, Strong, TD, Cooke, CA, El-Haddad, H, Tuder, RM, Berkowitz, DE & Champion, HC 2011, 'Phosphodiesterase-5A (PDE5A) is localized to the endothelial caveolae and modulates NOS3 activity', Cardiovascular Research, vol. 90, no. 2, pp. 353-363. https://doi.org/10.1093/cvr/cvq410
Gebska, Milena A. ; Stevenson, Blake K. ; Hemnes, Anna R. ; Bivalacqua, Trinity ; Haile, Azeb ; Hesketh, Geoffrey G. ; Murray, Christopher I. ; Zaiman, Ari L. ; Halushka, Marc K ; Krongkaew, Nispa ; Strong, Travis D. ; Cooke, Carol A. ; El-Haddad, Hazim ; Tuder, Rubin M. ; Berkowitz, Dan E ; Champion, Hunter C. / Phosphodiesterase-5A (PDE5A) is localized to the endothelial caveolae and modulates NOS3 activity. In: Cardiovascular Research. 2011 ; Vol. 90, No. 2. pp. 353-363.
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AU - Stevenson, Blake K.

AU - Hemnes, Anna R.

AU - Bivalacqua, Trinity

AU - Haile, Azeb

AU - Hesketh, Geoffrey G.

AU - Murray, Christopher I.

AU - Zaiman, Ari L.

AU - Halushka, Marc K

AU - Krongkaew, Nispa

AU - Strong, Travis D.

AU - Cooke, Carol A.

AU - El-Haddad, Hazim

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AU - Berkowitz, Dan E

AU - Champion, Hunter C.

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KW - Primary human endothelial cells

KW - Pulmonary circulation

KW - Signalling pathway

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