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Phosphodiesterase 9A controls nitric-oxide-independent cGMP and hypertrophic heart disease.

Lee DI, Zhu G, Sasaki T, Cho GS, Hamdani N, Holewinski R, Jo SH, Danner T, Zhang M, Rainer PP, Bedja D, Kirk JA, Ranek MJ, Dostmann WR, Kwon C, Margulies KB, Van Eyk JE, Paulus WJ, Takimoto E, Kass DA - Nature (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

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

ABSTRACT
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 disease. However, cGMP stimulation alone is limited by counter-adaptions including PDE upregulation. Furthermore, although PDE5A regulates nitric-oxide-generated cGMP, nitric oxide signalling is often depressed by heart disease. 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.

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Chronic PDE9A-inhibition reverses pre-established hypertrophy/dysfunction in NOS-independent mannera, M-mode echocardiograms from mice ±L-NAME exposed to 5-wk TAC±PDE5 or PDE9-inhibition starting 1-week after TAC. b, PF-9613 and SIL reversed cardiac dysfunction from TAC in L-NAME(−), but only PF-9613 was effective in L-NAME(+) mice; *-p<0.01;#-p<0.05-vs-TAC. c, cyclic-GMP levels and d, In vitro PKG activity measured in the same experiments. e, Transcription factor activation in RNCMs stimulated by PE±gene-silencing of PDE5A or PDE9A; (Symbols for panels c–e; *p<0.005,#-p<0.01,†p<0.001,$-p<0.05. f, Differential phosphorylation of target proteins following PDE9A versus PDE5A inhibition. L-NAME co-treatment reversed PDE5A-selective changes (red) far more than PDE9A-selective changes.
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Figure 4: Chronic PDE9A-inhibition reverses pre-established hypertrophy/dysfunction in NOS-independent mannera, M-mode echocardiograms from mice ±L-NAME exposed to 5-wk TAC±PDE5 or PDE9-inhibition starting 1-week after TAC. b, PF-9613 and SIL reversed cardiac dysfunction from TAC in L-NAME(−), but only PF-9613 was effective in L-NAME(+) mice; *-p<0.01;#-p<0.05-vs-TAC. c, cyclic-GMP levels and d, In vitro PKG activity measured in the same experiments. e, Transcription factor activation in RNCMs stimulated by PE±gene-silencing of PDE5A or PDE9A; (Symbols for panels c–e; *p<0.005,#-p<0.01,†p<0.001,$-p<0.05. f, Differential phosphorylation of target proteins following PDE9A versus PDE5A inhibition. L-NAME co-treatment reversed PDE5A-selective changes (red) far more than PDE9A-selective changes.

Mentions: All groups developed marked chamber dysfunction, dilation, and hypertrophy after 1-week TAC (pre-treatment; Fig. 4a, b). In L-NAME(−) mice, inhibiting either PDE reversed changes to near sham-control levels, lowered post-mortem LV mass, lung weight, and abnormal molecular signatures (Extended Data Fig. 8). However, in L-NAME(+) mice, only PDE9A inhibition was effective (Fig. 4a, b). Blocking either PDE increased myocardial cGMP in L-NAME(−)-TAC but only PDE9A-inhibition did so in L-NAME(+)-TAC mice (Fig. 4c). L-NAME prevented PKG activation by SIL but not PF-9613 (Fig. 4d). In vitro PKG activity in PDE9A treated myocardium was itself little altered. This parallels reported data with NP stimulation, but differing from increases seen with PDE5A inhibition5.


Phosphodiesterase 9A controls nitric-oxide-independent cGMP and hypertrophic heart disease.

Lee DI, Zhu G, Sasaki T, Cho GS, Hamdani N, Holewinski R, Jo SH, Danner T, Zhang M, Rainer PP, Bedja D, Kirk JA, Ranek MJ, Dostmann WR, Kwon C, Margulies KB, Van Eyk JE, Paulus WJ, Takimoto E, Kass DA - Nature (2015)

Chronic PDE9A-inhibition reverses pre-established hypertrophy/dysfunction in NOS-independent mannera, M-mode echocardiograms from mice ±L-NAME exposed to 5-wk TAC±PDE5 or PDE9-inhibition starting 1-week after TAC. b, PF-9613 and SIL reversed cardiac dysfunction from TAC in L-NAME(−), but only PF-9613 was effective in L-NAME(+) mice; *-p<0.01;#-p<0.05-vs-TAC. c, cyclic-GMP levels and d, In vitro PKG activity measured in the same experiments. e, Transcription factor activation in RNCMs stimulated by PE±gene-silencing of PDE5A or PDE9A; (Symbols for panels c–e; *p<0.005,#-p<0.01,†p<0.001,$-p<0.05. f, Differential phosphorylation of target proteins following PDE9A versus PDE5A inhibition. L-NAME co-treatment reversed PDE5A-selective changes (red) far more than PDE9A-selective changes.
© Copyright Policy - permissions-link
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4376609&req=5

Figure 4: Chronic PDE9A-inhibition reverses pre-established hypertrophy/dysfunction in NOS-independent mannera, M-mode echocardiograms from mice ±L-NAME exposed to 5-wk TAC±PDE5 or PDE9-inhibition starting 1-week after TAC. b, PF-9613 and SIL reversed cardiac dysfunction from TAC in L-NAME(−), but only PF-9613 was effective in L-NAME(+) mice; *-p<0.01;#-p<0.05-vs-TAC. c, cyclic-GMP levels and d, In vitro PKG activity measured in the same experiments. e, Transcription factor activation in RNCMs stimulated by PE±gene-silencing of PDE5A or PDE9A; (Symbols for panels c–e; *p<0.005,#-p<0.01,†p<0.001,$-p<0.05. f, Differential phosphorylation of target proteins following PDE9A versus PDE5A inhibition. L-NAME co-treatment reversed PDE5A-selective changes (red) far more than PDE9A-selective changes.
Mentions: All groups developed marked chamber dysfunction, dilation, and hypertrophy after 1-week TAC (pre-treatment; Fig. 4a, b). In L-NAME(−) mice, inhibiting either PDE reversed changes to near sham-control levels, lowered post-mortem LV mass, lung weight, and abnormal molecular signatures (Extended Data Fig. 8). However, in L-NAME(+) mice, only PDE9A inhibition was effective (Fig. 4a, b). Blocking either PDE increased myocardial cGMP in L-NAME(−)-TAC but only PDE9A-inhibition did so in L-NAME(+)-TAC mice (Fig. 4c). L-NAME prevented PKG activation by SIL but not PF-9613 (Fig. 4d). In vitro PKG activity in PDE9A treated myocardium was itself little altered. This parallels reported data with NP stimulation, but differing from increases seen with PDE5A inhibition5.

Bottom Line: 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.

View Article: PubMed Central - PubMed

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

ABSTRACT
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 disease. However, cGMP stimulation alone is limited by counter-adaptions including PDE upregulation. Furthermore, although PDE5A regulates nitric-oxide-generated cGMP, nitric oxide signalling is often depressed by heart disease. 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.

Show MeSH
Related in: MedlinePlus