Anti‐mitogenic effects of β‐agonists and PGE<sub>2</sub>on airway smooth muscle are PKA dependent

Hou, Yan; Deshpande, Deepak A.; Misior, Anna M.; Miles, Matthew; Saxena, Himansh; Riemer, Ellen C.; Pascual, Rodolfo M.; Panettieri, Reynold A.; Penn, Raymond B.

doi:10.1096/fj.10-164798

Cited by 58 publications

(78 citation statements)

References 29 publications

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“…Epac itself or together with its target Rap has been implicated in the control of cell proliferation, and another major cAMP target, protein kinase A (PKA), was shown to act synergistically with these proteins in this process in several cases. [52][53][54][55][56][57] Our observation underlines the link between Epac, Rap, PKA and the cell cycle. However, further investigations are needed to determine whether their effects in HUVECs are proliferative or anti-proliferative and Rap-or PKA-dependent.…”

Section: Cid-msa Etd and Hcd Have Complementary Contributions To Phosupporting

confidence: 54%

Quantitative global phosphoproteomics of human umbilical vein endothelial cells after activation of the Rap signaling pathway

et al. 2013

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The small GTPase Rap1 is required for proper cell-cell junction formation and also plays a key role in mediating cAMP-induced tightening of adherens junctions and subsequent increased barrier function of endothelial cells. To further study how Rap1 controls barrier function, we performed quantitative global phosphoproteomics in human umbilical vein endothelial cells (HUVECs) prior to and after Rap1 activation by the Epac-selective cAMP analog 8-pCPT-2 0 -O-Me-cAMP-AM (007-AM). Tryptic digests were labeled using stable isotope dimethyl labeling, enriched with phosphopeptides by strong cation exchange (SCX), followed by titanium(IV) immobilized metal affinity chromatography (Ti 4+ -IMAC) and analyzed by high resolution mass spectrometry. We identified 19 859 unique phosphopeptides containing 17 278 unique phosphosites on 4594 phosphoproteins, providing the largest HUVEC phosphoproteome to date. Of all identified phosphosites, 220 (B1%) were more than 1.5-fold up-or downregulated upon Rap activation, in two independent experiments. Compatible with the function of Rap1, these alterations were found predominantly in proteins regulating the actin cytoskeleton, cell-cell junctions and cell adhesion.

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Section: Cid-msa Etd and Hcd Have Complementary Contributions To Phosupporting

confidence: 54%

Quantitative global phosphoproteomics of human umbilical vein endothelial cells after activation of the Rap signaling pathway

et al. 2013

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“…Indeed, the accumulation of cAMP and of PKA during both acute and chronic with agonists were markedly greater with PGE2 compared with b 2 AR agonists (14,(25)(26)(27)(28) with regard to greater efficiency in the inhibition of HASM cell growth (26,29) and migration (6). Indeed, our published data show that, in contrast to b 2 AR agonists that transiently increase the phosphorylation of VASP but decrease in effectiveness by 3 hours, PGE2 promotes a PKA-dependent phosphorylation of VASP that is sustained for up to 18 hours, suggesting that prolonged exposure to PGE2 does not desensitize EP2 receptors (25). Such differences between b-agonists and PGE2 may be explained by differential mechanisms of b 2 AR and EP2 receptor desensitization in HASM cells.…”

Section: Discussionmentioning

confidence: 99%

“…Our previously published studies demonstrated that PGE2 promotes the cAMP-dependent activation of PKA and phosphorylation of VASP in HASM cells via the prostaglandin E2 (EP2) receptor (25,26), which, in contrast to b 2 AR, is relatively resistant to agonist-specific desensitization. Indeed, the accumulation of cAMP and of PKA during both acute and chronic with agonists were markedly greater with PGE2 compared with b 2 AR agonists (14,(25)(26)(27)(28) with regard to greater efficiency in the inhibition of HASM cell growth (26,29) and migration (6). Indeed, our published data show that, in contrast to b 2 AR agonists that transiently increase the phosphorylation of VASP but decrease in effectiveness by 3 hours, PGE2 promotes a PKA-dependent phosphorylation of VASP that is sustained for up to 18 hours, suggesting that prolonged exposure to PGE2 does not desensitize EP2 receptors (25).…”

Section: Discussionmentioning

confidence: 99%

β₂-Adrenergic Receptor Agonists Modulate Human Airway Smooth Muscle Cell Migration via Vasodilator-Stimulated Phosphoprotein

Goncharova

Goncharov

Zhao

et al. 2012

Am J Respir Cell Mol Biol

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Severe asthma manifests as airway remodeling and irreversible airway obstruction, in part because of the proliferation and migration of human airway smooth muscle (HASM) cells. We previously reported that cyclic adenosine monophosphate-mobilizing agents, including b 2 -adrenergic receptor (b 2 AR) agonists, which are mainstay of asthma therapy, and prostaglandin E2 (PGE2), inhibit the migration of HASM cells, although the mechanism for this migration remains unknown. Vasodilator-stimulated phosphoprotein (VASP), an anticapping protein, modulates the formation of actin stress fibers during cell motility, and is negatively regulated by protein kinase A (PKA)-specific inhibitory phosphorylation at serine 157 (Ser157). Here, we show that treatment with b 2 AR agonists and PGE2 induces the PKA-dependent phosphorylation of VASP and inhibits the migration of HASM cells. The stable expression of PKA inhibitory peptide and the small interfering (si) RNA-induced depletion of VASP abolish the inhibitory effects of albuterol and PGE2 on the migration of HASM cells. Importantly, prolonged treatment with albuterol prevents the agonist-induced phosphorylation of VASP at Ser157, and reverses the inhibitory effects of albuterol and formoterol, but not PGE2, on the basal and PDGF-induced migration of HASM cells. Collectively, our data demonstrate that b 2 AR agonists selectively inhibit the migration of HASM cells via a b 2 AR/PKA/ VASP signaling pathway, and that prolonged treatment with albuterol abolishes the inhibitory effect of b-agonists on the phosphorylation of VASP and migration of HASM cells because of b 2 AR desensitization.Keywords: airway hyperresponsiveness; b 2 -adrenergic receptor desensitization; protein kinase A; albuterol; formoterol Human airway smooth muscle (HASM) hyperplasia and remodeling are characteristic features of airways that contribute to their irreversible obstruction in patients with severe asthma. Accumulating evidence suggests that cell migration contributes to airway smooth muscle hyperplasia and remodeling (1, 2). Recently, the migration of airway smooth muscle (ASM) cells toward the epithelium and lumen of the airway was reported as a potential contributor to the increased smooth muscle mass in patients with asthma (3, 4). Further, cell-culture studies show that mitogens and inflammatory mediators involved in asthma pathogenesis and pharmacological agents in current used for the treatment of asthma modulate the migration of ASM cells (1,5,6). Taken together, these studies suggest that the migration of ASM cells contributes to ASM cell hyperplasia under asthmarelated conditions (1). Although the role of HASM hyperplasia in asthma is well established, the molecular mechanisms regulating the migration of HASM cells under asthma-related conditions are poorly understood (1).Evidence demonstrates that cyclic adenosine monophosphate (cAMP)-inducing agents, including the long-acting b 2 -adrenergic receptor (b 2 AR) agonist formoterol, inhibit the agonist-dependent migration of ASM cells (5, 7). We p...

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“…Abnormal smooth muscle cell division contributes to the progression of airway remodeling, a key characteristic of asthma (1,2).…”

Section: Clinical Relevancementioning

confidence: 99%

c-Abl Tyrosine Kinase Regulates Cytokinesis of Human Airway Smooth Muscle Cells

Chen

Tang

2014

Am J Respir Cell Mol Biol

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Cytokinesis is a critical step of airway smooth muscle cell division that plays an essential role in the development and homeostasis of the respiratory system, as well as the progression of airway remodeling. The mechanisms that regulate smooth muscle cytokinesis are not fully understood. c-Abl (c-Abelson tyrosine kinase) is a nonreceptor protein tyrosine kinase that has a role in regulating actin dynamics and smooth muscle contraction. The role of c-Abl in cytokinesis has not been investigated. Here, c-Abl was found in the contractile ring, as evidenced by immunofluorescent microscopy. In addition, cortactin is a phosphorylatable protein that has been implicated in actin filament assembly. In this report, phosphorylated cortactin was also found in the contractile ring. Knockdown of c-Abl by RNA interference attenuated cortactin phosphorylation in the midzone and contractile ring formation. c-Abl knockdown decreased the number of cells undergoing cytokinesis, but increased the quantity of cells in metaphase/anaphase and the number of multinucleate cells. Treatment with the c-Abl pharmacological inhibitors, imatinib and GNF-5, had similar effects. Furthermore, the expression of a nonphosphorylatable cortactin mutant diminished cytokinesis. Finally, inhibition of actin filament assembly by latrunculin A attenuated c-Abl recruitment to the midzone. Thus, we propose a novel mechanism that regulates smooth muscle cell cytokinesis. c-Abl is recruited to the equator during cytokinesis, which may mediate cortactin phosphorylation. Phosphorylated cortactin may promote actin filament assembly, which facilitates contractile ring formation and cytokinesis. In addition, actin filament polymerization may facilitate the positioning of c-Abl to the contractile ring.

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Anti‐mitogenic effects of β‐agonists and PGE₂on airway smooth muscle are PKA dependent

Cited by 58 publications

References 29 publications

Quantitative global phosphoproteomics of human umbilical vein endothelial cells after activation of the Rap signaling pathway

Quantitative global phosphoproteomics of human umbilical vein endothelial cells after activation of the Rap signaling pathway

β₂-Adrenergic Receptor Agonists Modulate Human Airway Smooth Muscle Cell Migration via Vasodilator-Stimulated Phosphoprotein

c-Abl Tyrosine Kinase Regulates Cytokinesis of Human Airway Smooth Muscle Cells

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Anti‐mitogenic effects of β‐agonists and PGE2on airway smooth muscle are PKA dependent

Cited by 58 publications

References 29 publications

Quantitative global phosphoproteomics of human umbilical vein endothelial cells after activation of the Rap signaling pathway

Quantitative global phosphoproteomics of human umbilical vein endothelial cells after activation of the Rap signaling pathway

β2-Adrenergic Receptor Agonists Modulate Human Airway Smooth Muscle Cell Migration via Vasodilator-Stimulated Phosphoprotein

c-Abl Tyrosine Kinase Regulates Cytokinesis of Human Airway Smooth Muscle Cells

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Anti‐mitogenic effects of β‐agonists and PGE₂on airway smooth muscle are PKA dependent

β₂-Adrenergic Receptor Agonists Modulate Human Airway Smooth Muscle Cell Migration via Vasodilator-Stimulated Phosphoprotein