Targeting the heme-oxidized nitric oxide receptor for selective vasodilatation of diseased blood vessels
Johannes-Peter Stasch, … , Werner Müller-Esterl, Harald H.H.W. Schmidt
Johannes-Peter Stasch, … , Werner Müller-Esterl, Harald H.H.W. Schmidt
Published September 1, 2006
Citation Information: J Clin Invest. 2006;116(9):2552-2561. https://doi.org/10.1172/JCI28371.
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Research Article Cardiology

Targeting the heme-oxidized nitric oxide receptor for selective vasodilatation of diseased blood vessels

Abstract

ROS are a risk factor of several cardiovascular disorders and interfere with NO/soluble guanylyl cyclase/cyclic GMP (NO/sGC/cGMP) signaling through scavenging of NO and formation of the strong oxidant peroxynitrite. Increased oxidative stress affects the heme-containing NO receptor sGC by both decreasing its expression levels and impairing NO-induced activation, making vasodilator therapy with NO donors less effective. Here we show in vivo that oxidative stress and related vascular disease states, including human diabetes mellitus, led to an sGC that was indistinguishable from the in vitro oxidized/heme-free enzyme. This sGC variant represents what we believe to be a novel cGMP signaling entity that is unresponsive to NO and prone to degradation. Whereas high-affinity ligands for the unoccupied heme pocket of sGC such as zinc–protoporphyrin IX and the novel NO-independent sGC activator 4-[((4-carboxybutyl){2-[(4-phenethylbenzyl)oxy]phenethyl}amino) methyl [benzoic]acid (BAY 58-2667) stabilized the enzyme, only the latter activated the NO-insensitive sGC variant. Importantly, in isolated cells, in blood vessels, and in vivo, BAY 58-2667 was more effective and potentiated under pathophysiological and oxidative stress conditions. This therapeutic principle preferentially dilates diseased versus normal blood vessels and may have far-reaching implications for the currently investigated clinical use of BAY 58-2667 as a unique diagnostic tool and highly innovative vascular therapy.

Authors

Johannes-Peter Stasch, Peter M. Schmidt, Pavel I. Nedvetsky, Tatiana Y. Nedvetskaya, Arun Kumar H.S., Sabine Meurer, Martin Deile, Ashraf Taye, Andreas Knorr, Harald Lapp, Helmut Müller, Yagmur Turgay, Christiane Rothkegel, Adrian Tersteegen, Barbara Kemp-Harper, Werner Müller-Esterl, Harald H.H.W. Schmidt

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Figure 4

Effects of BAY 58-2667 on sGC protein levels under normal and oxidative conditions.

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Effects of BAY 58-2667 on sGC protein levels under normal and oxidative ...
(A) Endothelial cells were incubated for 24 hours with 10 μM ODQ, 10 μM BAY 58-2667, or the combination of both compounds. (B) Endothelial cells were treated with different concentrations of BAY 58-2667 for 24 hours. (C) Endothelial cells were incubated for 24 hours with 10 μM methylene blue (MB), 10 μM BAY 58-2667, or the combination of both compounds. (D) Porcine smooth muscle cells were incubated for 24 hours with 10 μM ODQ, 10 μM BAY 58-2667, or the combination of both compounds. (E) Endothelial cells were incubated for 24 hours with 500 μM SIN-1, 10 μM BAY 58-2667, or the combination of both compounds. (F) Endothelial cells were treated with different concentrations of Zn-PPIX for 48 hours. (G) sGC α and (H) sGC β protein expression in endothelial cells preincubated with 10 μM ODQ, 10 μg/ml cycloheximide, or 10 μg/ml emetine for 0, 2, 4, and 8 hours. (I) Endothelial cells were treated for 24 hours with 250 μM 8-Br-cGMP, 100 μM YC-1, 200 μM of the PDE inhibitor Zaprinast, and 100 μM DEA/NO. All sGC protein levels were determined by Western blot. Data are expressed as percent of control (means ± SEM of 3–6 independent experiments performed in triplicate). *P < 0.05; **P < 0.01. On A, C, D, and E, representative blots are shown.