Identification of Potent and Selective Amidobipyridyl Inhibitors of Protein Kinase D

Meredith, Erik; Beattie, Kimberly; Burgis, Robin; Capparelli, Michael; Chapo, Joseph; DiPietro, Lucian; Gamber, Gabriel G.; Enyedy, Istvan; Hood, David B.; Hosagrahara, Vinayak; Jewell, Charles; Koch, Keith A.; Lee, Wendy; Lemon, Douglas D.; McKinsey, Timothy A.; Miranda, Karl; Pagratis, Nikos; Phan, Dillon; Plato, Craig F.; Rao, Chang; Rozhitskaya, Olga; Soldermann, Nicolas; Springer, Clayton; Eis, Maurice van; Vega, Richard B.; Yan, Wanlin; Zhu, Qingyu; Monovich, Lauren G.

doi:10.1021/jm100076w

Cited by 39 publications

(49 citation statements)

References 30 publications

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“…Furthermore, phosphorylation of these residues, which flank a nuclear localization signal domain in both HDAC4 and HDAC5, has been causally implicated in the neurohormonal induction of HDAC nuclear export and thereby the disinhibition of MEF2-driven transcriptional reprogramming that facilitates myocyte hypertrophy. 4,26 We therefore explored the individual regulation of HDAC4 and HDAC5 in ARVM through heterologous expression of their FLAG epitope-tagged variants (FLAG-HDAC4 and FLAG-HDAC5), using a newly described selective PKD inhibitor (identified as compound 12a 27 or bipyridyl PKD inhibitor [BPKDi] 28 ) to confirm the role of PKD activity in their ET1-induced phosphorylation. In an initial step, we extended the recent pharmacological characterization of BPKDi 27,28 by screening it against a larger panel of Ͼ120 protein kinases using validated in vitro kinase assays, 21 which underscored the inhibitor's remarkable selectivity for PKD (Online Table I).…”

Section: Resultsmentioning

confidence: 99%

“…4,26 We therefore explored the individual regulation of HDAC4 and HDAC5 in ARVM through heterologous expression of their FLAG epitope-tagged variants (FLAG-HDAC4 and FLAG-HDAC5), using a newly described selective PKD inhibitor (identified as compound 12a 27 or bipyridyl PKD inhibitor [BPKDi] 28 ) to confirm the role of PKD activity in their ET1-induced phosphorylation. In an initial step, we extended the recent pharmacological characterization of BPKDi 27,28 by screening it against a larger panel of Ͼ120 protein kinases using validated in vitro kinase assays, 21 which underscored the inhibitor's remarkable selectivity for PKD (Online Table I). We also confirmed the ability of BPKDi to inhibit PKD activity in ARVM at concentrations Ͼ1 mol/L (Online Figure I), with the lower potency reflecting the higher ATP concentration in cells relative to that used for in vitro kinase assays, as observed also for other ATP-competitive kinase inhibitors.…”

Section: Resultsmentioning

confidence: 99%

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Neurohormonal Regulation of Cardiac Histone Deacetylase 5 Nuclear Localization by Phosphorylation-Dependent and Phosphorylation-Independent Mechanisms

Haworth

Σταθοπούλου²,

Candasamy

et al. 2012

Circulation Research

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Rationale: Myocyte enhancer factor 2 (MEF2) transcription factors drive the genetic reprogramming that precipitates pathological cardiac hypertrophy and remodeling. Class II histone deacetylase (HDAC) isoforms, such as HDAC5, act as signal-responsive repressors of MEF2 activity in cardiac myocytes and their nuclear export provides a key mechanism for the neurohormonal induction of such activity.Objective: To delineate the mechanism(s) through which 2 clinically relevant neurohormonal stimuli, endothelin-1 (ET1) and the ␤-adrenergic receptor (␤-AR) agonist isoproterenol (ISO), may regulate HDAC5 nuclear localization in adult cardiac myocytes. Methods and Results: ET1 induced HDAC5 phosphorylation and nuclear export in ventricular myocytes fromthe adult rat heart. Use of a novel, highly selective protein kinase D (PKD) inhibitor and a nonphosphorylatable HDAC5 mutant revealed that PKD-mediated phosphorylation was necessary for ET1-induced HDAC5 nuclear export. In contrast, ISO reduced HDAC5 phosphorylation in the presence or absence of ET1 but still induced HDAC5 nuclear export. ISO-induced HDAC5 nuclear export occurred through a ␤ 1 -AR-mediated oxidative process that was independent of PKD, protein kinase A, and Ca 2؉ /calmodulin-dependent kinase II activities. Although ET1 and ISO shared a similar ability to induce HDAC5 nuclear export, albeit through distinct phosphorylation-dependent versus phosphorylation-independent mechanisms, ISO induced a significantly greater increase in MEF2 activity.Conclusions: PKD-mediated HDAC5 phosphorylation and nuclear export are unlikely to be of major importance in regulating MEF2-driven cardiac remodeling in the presence of sympathetic activity with intact ␤ 1 -AR signaling, which would not only counteract HDAC5 phosphorylation but also induce HDAC5 nuclear export through a novel phosphorylation-independent, oxidation-mediated mechanism. Inhibition of this mechanism may contribute to the therapeutic efficacy of ␤ 1 -AR antagonists in heart failure. (Circ Res. 2012;110:1585-1595.) Key Words: cardiac hypertrophy Ⅲ histone deacetylases Ⅲ myocyte enhancer factor 2 Ⅲ protein kinases Ⅲ signal transduction A substantial body of evidence indicates that the activity of myocyte enhancer factor 2 (MEF2) transcription factors is a critical driver of the pathological cardiac hypertrophy and remodeling processes that culminate in heart failure, in both acquired 1 and genetic 2 forms of the disease. An extensive series of studies in recent years, particularly in the Olson laboratory, have revealed that class II histone deacetylase (HDAC) isoforms, such as HDAC4 and HDAC5, act as signal-responsive repressors of nuclear MEF2 activity in cardiac myocytes and that their spatial regulation provides a key mechanism for the neurohormonal control of such activity. 3,4 The compelling scheme that has emerged is that HDAC4 and HDAC5 are direct substrates for serine/threonine protein kinases of the Ca 2ϩ /calmodulin-dependent kinase (CaMK) superfamily, such as CaMKII 5 and protein kinase D (PKD), 6 an...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Neurohormonal Regulation of Cardiac Histone Deacetylase 5 Nuclear Localization by Phosphorylation-Dependent and Phosphorylation-Independent Mechanisms

Haworth

Σταθοπούλου²,

Candasamy

et al. 2012

Circulation Research

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“…Inhibitors targeting PKD1 such as BPDKi have been developed but this aminobipyridyl compound failed to blunt animal cardiac hypertrophy (31). Our new results call for further tests on PKD3-selective inhibitors or pan-PKD inhibitors to determine their efficacy in blunting cardiac hypertrophy and heart failure.…”

Section: Discussionmentioning

confidence: 97%

“…Mice with PKD1 ablation show substantial resistance to cardiac hypertrophy (30). Unexpectedly, a recent study reported a newly developed PKD1-selective inhibitor failed to attenuate PCH in whole animal models (31). One possible explanation for such a discrepancy is that PKD3 could substitute for PKD1 as a HDAC5 kinase as documented in non-cardiac cells (32).…”

Section: Pathological Cardiac Hypertrophy (Pch)mentioning

confidence: 99%

Protein Kinase D3 Is a Pivotal Activator of Pathological Cardiac Hypertrophy by Selectively Increasing the Expression of Hypertrophic Transcription Factors

Cai

et al. 2011

Journal of Biological Chemistry

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Background: Post-translational modulation of preexisting translational factors is a well-established cardiac hypertrophic mechanism. Results: The abundance of NFATc4, Nkx2.5 and GATA4 is up-regulated by newly-expressed protein kinase D3 (PKD3) to induce pathological cardiac hypertrophy. Conclusion: PKD3 is a pivotal mediator of cardiac hypertrophic signaling cascade. Significance: PKD3 is a potential new drug target for pathological cardiac hypertrophy.

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“…HDAC subtype-specific inhibition might be a way to resolve this benefit conflict, but targeting upstream of HDACs may hold unique benefits. Along these lines, cardiac-specific deletion of PKD1 (and CaMKII␦ knock-out) limit cardiac remodeling after aortic constriction (5,22) and novel PKD inhibitors have had some success in the prevention of pathological hypertrophy (23)(24)(25) as well as of pancreatic and prostatic cancer growth (17,26). In the heart, a better mechanistic understanding of how cardiomyocyte PKD and CaMK are activated in response to pathogenic neurohumoral stimuli (5-6) is needed to evaluate the therapeutic potential of specific PKD/ CaMKII inhibition.…”

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confidence: 99%

Spatiotemporally Distinct Protein Kinase D Activation in Adult Cardiomyocytes in Response to Phenylephrine and Endothelin

Bossuyt

Chang

Helmstadter

et al. 2011

Journal of Biological Chemistry

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Protein kinase D (PKD) is a nodal point in cardiac hypertrophic signaling. It triggers nuclear export of class II histone deacetylase (HDAC) and regulates transcription. Although this pathway is thought to be critical in cardiac hypertrophy and heart failure, little is known about spatiotemporal aspects of PKD activation at the myocyte level. Here, we demonstrate that in adult cardiomyocytes two important neurohumoral stimuli that induce hypertrophy, endothelin-1 (ET1) and phenylephrine (PE), trigger comparable global PKD activation and HDAC5 nuclear export, but via divergent spatiotemporal PKD signals. PE-induced HDAC5 export is entirely PKD-dependent, involving fleeting sarcolemmal PKD translocation (for activation) and very rapid subsequent nuclear import. In contrast, ET1 recruits and activates PKD that remains predominantly sarcolemmal. This explains why PE-induced nuclear HDAC5 export in myocytes is totally PKD-dependent, whereas ET1-induced HDAC5 export depends more prominently on InsP 3 and CaMKII signaling. Thus ␣-adrenergic and ET-1 receptor signaling via PKD in adult myocytes feature dramatic differences in cellular localization and translocation in mediating hypertrophic signaling. This raises new opportunities for targeted therapeutic intervention into distinct limbs of this hypertrophic signaling pathway.Various stresses trigger cardiac hypertrophy, remodeling, and functional alterations (1, 2). Prolonged stress can also become maladaptive, leading to heart failure, cardiac arrhythmias, and sudden death. Many of these changes are mediated by altered gene expression, and Class II histone deacetylases (HDACs) 3 (e.g. HDAC5) are recognized as key modulators of this genetic reprogramming. HDAC5 represses transcription by promoting more condensed DNA, and represses transcription factors such as myocyte enhancing factor 2 (MEF2). HDAC5 phosphorylation triggers its nuclear export (allowing gene activation (Fig. 1a)). Both protein kinase D (PKD) and calmodulin-dependent protein kinase II (CaMKII) are key HDAC kinases (1, 2) and accumulating evidence indicates both kinases are more active in heart failure and can contribute directly to cardiac pathogenesis (3-6). Cardiac PKD is activated in response to hypertension, pressure overload, and chronic neurohumoral signaling (7). Overexpression of constitutively active PKD (and CaMKII␦) cause cardiac hypertrophy followed by chamber dilation (8, 9). Moreover there is increased expression of PKD (and CaMKII␦) in failing rat, rabbit, and human myocardium (8, 10). Thus, whereas PKD and CaMKII activation may be involved in a wide variety of cell functions (11-14), they are attractive potential therapeutic targets in cardiac disease. Therapeutic benefit of PKD/CaMKII inhibition may be largely due to prevention of HDAC (class II) phosphorylation, thereby maintaining the repressive effects of HDAC on transcription. Indeed, HDAC5 knock-out mice develop profound pathological cardiac hypertrophy and HDAC5 overexpression can limit progression of cardiac hypertrophy (15,16)...

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Identification of Potent and Selective Amidobipyridyl Inhibitors of Protein Kinase D

Cited by 39 publications

References 30 publications

Neurohormonal Regulation of Cardiac Histone Deacetylase 5 Nuclear Localization by Phosphorylation-Dependent and Phosphorylation-Independent Mechanisms

Neurohormonal Regulation of Cardiac Histone Deacetylase 5 Nuclear Localization by Phosphorylation-Dependent and Phosphorylation-Independent Mechanisms

Protein Kinase D3 Is a Pivotal Activator of Pathological Cardiac Hypertrophy by Selectively Increasing the Expression of Hypertrophic Transcription Factors

Spatiotemporally Distinct Protein Kinase D Activation in Adult Cardiomyocytes in Response to Phenylephrine and Endothelin

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