Decreased Cyclic GMP-protein Kinase G signaling impairs Angiogenesis in a Lamb Model of Persistent Pulmonary Hypertension of Newborn

The activity of glucose-6-phosphate dehydrogenase (G6PD) controls a vascular smooth muscle relaxing mechanism promoted by the oxidation of cytosolic NADPH, which has been associated with activation of the 1α form of protein kinase G (PKG-1α) by a thiol oxidation-elicited subunit dimerization. This PKG-1α-activation mechanism appears to contribute to responses of isolated endothelium-removed bovine pulmonary arteries (BPA) elicited by peroxide, cytosolic NADPH oxidation resulting from G6PD inhibition, and hypoxia. Dehydroepiandrosterone (DHEA) is a steroid hormone with pulmonary vasodilator activity, which has beneficial effects in treating pulmonary hypertension. Because multiple mechanisms have been suggested for the vascular effects of DHEA and one of the known actions of DHEA is inhibiting G6PD, we investigated whether it promoted relaxation associated with NADPH oxidation, PKG-1α dimerization, and PKG activation detected by increased vasodilator-stimulated phosphoprotein (VASP) phosphorylation. Relaxation of BPA to DHEA under aerobic or hypoxic conditions was associated with NADPH oxidation, PKG-1α dimerization, and increased VASP phosphorylation. The vasodilator activity of DHEA was markedly attenuated in pulmonary arteries and aorta from a PKG knockin mouse containing a serine in place of a cysteine involved in PKG dimerization. DHEA promoted increased PKG dimerization in lungs from wild-type mice, which was not detected in the PKG knockin mouse model. Thus PKG-1α dimerization is a major contributing factor to the vasodilator actions of DHEA and perhaps its beneficial effects in treating pulmonary hypertension.

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Endogenous Nitric Oxide Generation Linked to Ryanodine Receptors Activates Cyclic GMP / Protein Kinase G Pathway for Cell Proliferation of Neural Stem/Progenitor Cells Derived From Embryonic Hippocampus

Journal of Pharmacological Sciences ◽

10.1254/jphs.10290fp ◽

2011 ◽

Vol 115 (2) ◽

pp. 182-195 ◽

Cited By ~ 20

Author(s):

Masanori Yoneyama ◽

Koichi Kawada ◽

Tatsuo Shiba ◽

Kiyokazu Ogita

Keyword(s):

Nitric Oxide ◽

Cell Proliferation ◽

Protein Kinase ◽

Progenitor Cells ◽

Cyclic Gmp ◽

Ryanodine Receptors ◽

Protein Kinase G ◽

Neural Stem Progenitor Cells ◽

Endogenous Nitric Oxide

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Protein kinase G deficiency induces pulmonary hypertension in mice and humans

The FASEB Journal ◽

10.1096/fasebj.24.1_supplement.1023.5 ◽

2010 ◽

Vol 24 (S1) ◽

Author(s):

You‐Yang Zhao ◽

Muhammad K Mirza ◽

Sophie Sun ◽

Kristen Wasiukanis ◽

Stephen M Vogel ◽

...

Keyword(s):

Pulmonary Hypertension ◽

Protein Kinase ◽

Protein Kinase G

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Abstract 556: The Soluble Guanylyl Cyclase Activator Bay 60-2770 Inhibits Arterial Smooth Muscle Cell Migration in Protein Kinase G-dependent Manner

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.36.suppl_1.556 ◽

2016 ◽

Vol 36 (suppl_1) ◽

Author(s):

Andrew Holt ◽

Danielle Martin ◽

Patti Shaver ◽

Shaquria Adderley ◽

Joshua Stone ◽

...

Keyword(s):

Smooth Muscle ◽

Cell Migration ◽

Protein Kinase ◽

Cyclic Gmp ◽

Guanylyl Cyclase ◽

Soluble Guanylyl Cyclase ◽

Protein Kinase G ◽

Growth Disorders ◽

Arterial Smooth Muscle ◽

Cyclase Activator

Atherosclerotic lower extremity peripheral artery disease (PAD) is among the most prevalent, morbid and mortal of all cardiovascular disorders. Pathologic arterial smooth muscle (ASM) cell migration is a major component of atherogenic PAD and efforts aimed at attenuating its progression are clinically essential. Cyclic nucleotide signaling has long been studied for its growth-mitigating properties in the setting of PAD and other vascular growth disorders. In this study we hypothesized that the novel, heme-independent soluble guanylyl cyclase activator BAY 60-2770 (BAY) inhibits ASM cell migration through phosphorylation of the protein kinase G (PKG) target and actin-binding protein vasodilator-stimulated phosphoprotein (VASP). In a rat model of injury-induced arterial growth, BAY significantly reduced neointima formation and luminal narrowing compared to vehicle (Veh)-treated control arteries after 2 weeks. Using rat and human ASM cells BAY significantly attenuated cell migration, reduced G:F actin, and increased cyclic GMP content, PKG activity and phosphorylated VASP at Ser239 (pVASP.S239) compared to Veh controls. Using site-directed mutagenesis, both full-length VASP-overexpressing (wild type, WT) and VASP.S239 phosphorylation-resistant mutants showed significantly reduced cell migration compared to naïve controls, however, there was no effect on cell migration between either VASP transfected group in the presence of BAY. Interestingly, both VASP mutants showed significantly increased PKG activity compared to naïve cells, and in turn pharmacologic PKG blockade in the presence of BAY fully reversed the inhibitory effect of BAY alone on cell migration. These data suggest BAY has capacity to inhibit ASM cell migration through cyclic GMP/PKG/VASP signaling yet through mechanisms independent of pVASP.S239. Findings from this study implicate BAY via cyclic GMP/PKG/VASP as a potential pharmacotherapeutic agent against aberrant ASM growth disorders such as PAD.

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