Cyclic guanosine monophosphate (cGMP) is a second messenger molecule that transduces nitric-oxide- and natriuretic-peptide-coupled signalling, stimulating phosphorylation changes by protein kinase G. Enhancing cGMP synthesis or blocking its degradation by phosphodiesterase type 5A (PDE5A) protects against cardiovascular disease1,2. However, cGMP stimulation alone is limited by counter-adaptions including PDE upregulation3. Furthermore, although PDE5A regulates nitric-oxide-generated cGMP4,5, nitric oxide signalling is often depressed by heart disease6. PDEs controlling natriuretic-peptide-coupled cGMP remain uncertain. Here we show that cGMP-selective PDE9A (refs 7, 8) is expressed in the mammalian heart, including humans, and is upregulated by hypertrophy and cardiac failure. PDE9A regulates natriuretic-peptide- rather than nitric-oxide-stimulated cGMP in heart myocytes and muscle, and its genetic or selective pharmacological inhibition protects against pathological responses to neurohormones, and sustained pressure-overload stress. PDE9A inhibition reverses pre-established heart disease independent of nitric oxide synthase (NOS) activity, whereas PDE5A inhibition requires active NOS. Transcription factor activation and phosphoproteome analyses of myocytes with each PDE selectively inhibited reveals substantial differential targeting, with phosphorylation changes from PDE5A inhibition being more sensitive to NOS activation. Thus, unlike PDE5A, PDE9A can regulate cGMP signalling independent of the nitric oxide pathway, and its role in stress-induced heart disease suggests potential as a therapeutic target.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from $8.99

All prices are NET prices.

Change history

  • Corrected online 25 March 2015

    The received date year was corrected.


  1. 1.

    et al. Chronic inhibition of cyclic GMP phosphodiesterase 5A prevents and reverses cardiac hypertrophy. Nature Med. 11, 214–222 (2005)

  2. 2.

    , & Cyclic guanosine monophosphate signaling and phosphodiesterase-5 inhibitors in cardioprotection. J. Am. Coll. Cardiol. 59, 1921–1927 (2012)

  3. 3.

    , , & In vivo reconstitution of the negative feedback in nitric oxide/cGMP signaling: role of phosphodiesterase type 5 phosphorylation. Mol. Biol. Cell 15, 4023–4030 (2004)

  4. 4.

    , , & Cyclic guanosine monophosphate compartmentation in rat cardiac myocytes. Circulation 113, 2221–2228 (2006)

  5. 5.

    et al. Compartmentalization of cardiac β-adrenergic inotropy modulation by phosphodiesterase type 5. Circulation 115, 2159–2167 (2007)

  6. 6.

    , , , & Nitric oxide synthases in heart failure. Antioxid. Redox Signal. 18, 1078–1099 (2013)

  7. 7.

    , & Identification and characterization of a novel family of cyclic nucleotide phosphodiesterases. J. Biol. Chem. 273, 15553–15558 (1998)

  8. 8.

    , , , & Isolation and characterization of PDE9A, a novel human cGMP-specific phosphodiesterase. J. Biol. Chem. 273, 15559–15564 (1998)

  9. 9.

    & Biochemistry and physiology of cyclic nucleotide phosphodiesterases: essential components in cyclic nucleotide signaling. Annu. Rev. Biochem. 76, 481–511 (2007)

  10. 10.

    et al. Phosphodiesterase 9A regulates central cGMP and modulates responses to cholinergic and monoaminergic perturbation in vivo. J. Pharmacol. Exp. Ther. 341, 396–409 (2012)

  11. 11.

    , & Phosphodiesterase inhibitors as a target for cognition enhancement in aging and Alzheimer’s disease: a translational overview. Curr. Pharm. Des. 21, 317–331 (2015)

  12. 12.

    & Heart failure with preserved ejection fraction: mechanisms, clinical features, and therapies. Circ. Res. 115, 79–96 (2014)

  13. 13.

    , , , & Differential patterning of cGMP in vascular smooth muscle cells revealed by single GFP-linked biosensors. Proc. Natl Acad. Sci. USA 105, 365–370 (2008)

  14. 14.

    , , , & Novel snake venom ligand dendroaspis natriuretic peptide is selective for natriuretic peptide receptor-A in human heart: downregulation of natriuretic peptide receptor-A in heart failure. Circ. Res. 99, 183–190 (2006)

  15. 15.

    , , , & TRPC6-dependent pathway for myofibroblast transdifferentiation and wound healing in vivo. Dev. Cell 23, 705–715 (2012)

  16. 16.

    et al. Cyclic GMP/PKG-dependent inhibition of TRPC6 channel activity and expression negatively regulates cardiomyocyte NFAT activation: novel mechanism of cardiac stress modulation by PDE5 inhibition. J. Mol. Cell. Cardiol. 48, 713–724 (2010)

  17. 17.

    et al. Inhibition of TRPC6 channel activity contributes to the antihypertrophic effects of natriuretic peptides-guanylyl cyclase-A signaling in the heart. Circ. Res. 106, 1849–1860 (2010)

  18. 18.

    et al. cGMP signals modulate cAMP levels in a compartment-specific manner to regulate catecholamine-dependent signaling in cardiac myocytes. Circ. Res. 108, 929–939 (2011)

  19. 19.

    & Role of cyclic GMP in gene regulation. Front. Biosci. 10, 1239–1268 (2005)

  20. 20.

    et al. Cardiac-specific deletion of Gata4 reveals its requirement for hypertrophy, compensation, and myocyte viability. Circ. Res. 98, 837–845 (2006)

  21. 21.

    et al. Advances in targeting cyclic nucleotide phosphodiesterases. Nature Rev. Drug Discov. 13, 290–314 (2014)

  22. 22.

    et al. The cGMP signaling pathway as a therapeutic target in heart failure with preserved ejection fraction. J. Am Heart Assoc. 2, e000536 (2013)

  23. 23.

    et al. Effect of phosphodiesterase-5 inhibition on exercise capacity and clinical status in heart failure with preserved ejection fraction: a randomized clinical trial. J. Am. Med. Assoc. 309, 1268–1277 (2013)

  24. 24.

    et al. Low myocardial protein kinase G activity in heart failure with preserved ejection fraction. Circulation 126, 830–839 (2012)

  25. 25.

    et al. Angiotensin-neprilysin inhibition versus enalapril in heart failure. N. Engl. J. Med. 993–1004 (2014)

  26. 26.

    & Designer natriuretic peptides: a vision for the future of heart failure therapeutics. Can. J. Physiol. Pharmacol. 89, 593–601 (2011)

  27. 27.

    et al. Combined TRPC3 and TRPC6 blockade by selective small-molecule or genetic deletion inhibits pathological cardiac hypertrophy. Proc. Natl Acad. Sci. USA 111, 1551–1556 (2014)

  28. 28.

    et al. Design and discovery of 6-[(3S,4S)-4-methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl]-1-(tetrahydro-2H-pyr an-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (PF-04447943), a selective brain penetrant PDE9A inhibitor for the treatment of cognitive disorders. J. Med. Chem. 55, 9045–9054 (2012)

  29. 29.

    et al. Epicardial progenitors contribute to the cardiomyocyte lineage in the developing heart. Nature 454, 109–113 (2008)

  30. 30.

    et al. Cardiac resynchronization sensitizes the sarcomere to calcium by reactivating GSK-3beta. J. Clin. Invest. 124, 129–139 (2014)

Download references


We thank students R. D. Wardlow and X. Hu for their assistance with some of the assays and studies. This research was supported by: the National Institutes of Health (NIH) (HL-119012, HL-089297, HL-07227), Fondation Leducq TransAtlantic Network of Excellence, The Peter Belfer Foundation, Abraham and Virginia Weiss Professorship (D.A.K.); HL-093432 (E.T.), American Heart Association (D.I.L.) and Max Kade Fellowship of the Austrian Academy of Sciences (P.P.R). Procurement of human heart tissue was enabled by grants from the National Institutes of Health (HL089847 and HL105993) to K.B.M. N.H. and W.J.P. were supported by the European Commission FP7 project 2010 Health (MEDIA; 261409). R.H. and J.E.V.E. were supported by The Johns Hopkins Innovation Proteomics Center in Heart Failure (NHLBI-HV-10-05 (2) and HHSN268201000032C). W.R.D. was supported by NIH grant HL68891 and the Totman Trust for Biomedical Research. We thank Pfizer and in particular C. Schmidt and R. Kleiman for providing the Pde9a−/− mouse and PF-04449613, and L. Jaffe at the University of Connecticut Health Center for providing the PDE9A antibody.

Author information


  1. Division of Cardiology, Department of Medicine, The Johns Hopkins Medical Institutions, Baltimore, Maryland 21205, USA

    • Dong I. Lee
    • , Guangshuo Zhu
    • , Gun-Sik Cho
    • , Ronald Holewinski
    • , Thomas Danner
    • , Manling Zhang
    • , Peter P. Rainer
    • , Djahida Bedja
    • , Jonathan A. Kirk
    • , Mark J. Ranek
    • , Chulan Kwon
    • , Jennifer E. Van Eyk
    • , Eiki Takimoto
    •  & David A. Kass
  2. Advanced Medical Research Laboratories, Research Division, Mitsubishi Tanabe Pharma Corporation, Yokohama, Kanagawa 227-0033, Japan

    • Takashi Sasaki
  3. Department of Physiology, Institute for Cardiovascular Research, VU University Medical Center, Van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands

    • Nazha Hamdani
    •  & Walter J. Paulus
  4. Heart Institute and Advanced Clinical Biosystems Research Institute, Cedar Sinai Medical Center, 8700 Beverly Blvd, AHSP A9229 Los Angeles, California 90048, USA

    • Ronald Holewinski
    •  & Jennifer E. Van Eyk
  5. Department of Physiology, Institute of Bioscience and Biotechnology, BK21 plus Graduate Program, Kangwon National University College of Medicine, Chuncheon 200-701, Korea

    • Su-Hyun Jo
  6. Department of Pharmacology, University of Vermont, Burlington, Vermont 05405, USA

    • Wolfgang R. Dostmann
  7. Department of Medicine, Division of Cardiovascular Medicine, Cardiovascular Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA

    • Kenneth B. Margulies


  1. Search for Dong I. Lee in:

  2. Search for Guangshuo Zhu in:

  3. Search for Takashi Sasaki in:

  4. Search for Gun-Sik Cho in:

  5. Search for Nazha Hamdani in:

  6. Search for Ronald Holewinski in:

  7. Search for Su-Hyun Jo in:

  8. Search for Thomas Danner in:

  9. Search for Manling Zhang in:

  10. Search for Peter P. Rainer in:

  11. Search for Djahida Bedja in:

  12. Search for Jonathan A. Kirk in:

  13. Search for Mark J. Ranek in:

  14. Search for Wolfgang R. Dostmann in:

  15. Search for Chulan Kwon in:

  16. Search for Kenneth B. Margulies in:

  17. Search for Jennifer E. Van Eyk in:

  18. Search for Walter J. Paulus in:

  19. Search for Eiki Takimoto in:

  20. Search for David A. Kass in:


D.I.L. and D.A.K. conceived and directed the project, designed experiments and prepared the manuscript. D.I.L. conducted most of the experiments and analysed the data. G.Z. helped with in vivo experiments. T.S., S.-H.J., T.D., M.Z. and P.P.R. conducted molecular biology experiments. G.-S.C. and C.K. carried out immunostaining and in situ hybridization analyses. N.H. and W.J.P. performed experiments for HFPEF and aortic stenosis human samples. M.J.R. and R.H. performed the experiments for phosphoproteomics and J.A.K. contributed to the data analysis. D.B. performed echocardiography and drug treatment. K.B.M. coordinated the human sample and data collection at the University of Pennsylvania, and W.R.D., J.E.V.E. and E.T. helped with data interpretation and presentation.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to David A. Kass.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Tables 1-3.

About this article

Publication history







By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.