Stimulators of the soluble guanylyl cyclase: promising functional insights from rare coding atherosclerosis-related GUCY1A3 variants.

Abstract

Stimulators of the soluble guanylyl cyclase (sGC) are emerging therapeutic agents in cardiovascular diseases. Genetic alterations of the GUCY1A3 gene, which encodes the α1 subunit of the sGC, are associated with coronary artery disease. Studies investigating sGC stimulators in subjects with CAD and carrying risk-related variants in sGC are, however, lacking. Here, we functionally investigate the impact of coding GUCY1A3 variants on sGC activity and the therapeutic potential of sGC stimulators in vitro. In addition to a known loss-of-function variant, eight coding variants in GUCY1A3 were cloned and expressed in HEK 293 cells. Protein levels and dimerization capability with the β1 subunit were analysed by immunoblotting and co-immunoprecipitation, respectively. All α1 variants found in MI patients dimerized with the β1 subunit. Protein levels were reduced by 72 % in one variant (p < 0.01). Enzymatic activity was analysed using cGMP radioimmunoassay after stimulation with a nitric oxide (NO) donor. Five variants displayed decreased cGMP production upon NO stimulation (p < 0.001). The addition of the sGC stimulator BAY 41-2272 increased cGMP formation in all of these variants (p < 0.01). Except for the variant leading to decreased protein level, cGMP amounts reached the wildtype NO-induced level after addition of BAY 41-2272. In conclusion, rare coding variants in GUCY1A3 lead to reduced cGMP formation which can be rescued by a sGC stimulator in vitro. These results might therefore represent the starting point for discovery of novel treatment strategies for patients at risk with coding GUCY1A3 variants.

• Authors: Jana Wobst, Simon von Ameln, Bernhard Wolf, Michael Wierer, Tan An Dang, Hendrik B Sager, Stephanie Tennstedt, Christian Hengstenberg, Doris Koesling, Andreas Friebe, Siegmund L Braun, Jeanette Erdmann, Heribert Schunkert, Thorsten Kessler
• Journal: Basic research in cardiology (Basic Res Cardiol) Vol. 111 Issue 4 Pg. 51 (07 2016) ISSN: 1435-1803 [Electronic] Germany
• PMID: 27342234 (Publication Type: Journal Article)

Chemical References

• 3-(4-Amino-5-cyclopropylpyrimidine-2-yl)-1-(2-fluorobenzyl)-1H-pyrazolo(3,4-b)pyridine
• GUCY1A1 protein, human
• Pyrazoles
• Pyridines
• Soluble Guanylyl Cyclase
• Cyclic GMP

Topics

• Adult
• Animals
• Coronary Artery Disease (genetics)
• Cyclic GMP (biosynthesis, genetics)
• Genetic Predisposition to Disease (genetics)
• HEK293 Cells
• Humans
• Immunoblotting
• Immunoprecipitation
• Mice
• Middle Aged
• Mutation, Missense
• Polymerase Chain Reaction
• Polymorphism, Single Nucleotide
• Pyrazoles (pharmacology)
• Pyridines (pharmacology)
• Radioimmunoassay
• Soluble Guanylyl Cyclase (genetics, metabolism)
• Young Adult