Phosphodiesterases Regulate BAY 41-2272-Induced VASP Phosphorylation in Vascular Smooth Muscle Cells.

BAY 41-2272 (BAY), a stimulator of soluble guanylyl cyclase, increases cyclic nucleotides and inhibits proliferation of vascular smooth muscle cells (VSMCs). In this study, we elucidated mechanisms of action of BAY in its regulation of vasodilator-stimulated phosphoprotein (VASP) with an emphasis on VSMC phosphodiesterases (PDEs). BAY alone increased phosphorylation of VASP(Ser239) and VASP(Ser157), respective indicators of PKG and PKA signaling. IBMX, a non-selective inhibitor of PDEs, had no effect on BAY-induced phosphorylation at VASP(Ser239) but inhibited phosphorylation at VASP(Ser157). Selective inhibitors of PDE3 or PDE4 attenuated BAY-mediated increases at VASP(Ser239) and VASP(Ser157), whereas PDE5 inhibition potentiated BAY-mediated increases only at VASP(Ser157). In comparison, 8Br-cGMP increased phosphorylation at VASP(Ser239) and VASP(Ser157) which were not affected by selective PDE inhibitors. In the presence of 8Br-cAMP, inhibition of either PDE4 or PDE5 decreased VASP(Ser239) phosphorylation and inhibition of PDE3 increased phosphorylation at VASP(Ser239), while inhibition of PDE3 or PDE4 increased and PDE5 inhibition had no effect on VASP(Ser157) phosphorylation. These findings demonstrate that BAY operates via cAMP and cGMP along with regulation by PDEs to phosphorylate VASP in VSMCs and that the mechanism of action of BAY in VSMCs is different from that of direct cyclic nucleotide analogs with respect to VASP phosphorylation and the involvement of PDEs. Given a role for VASP as a critical cytoskeletal protein, these findings provide evidence for BAY as a regulator of VSMC growth and a potential therapeutic agent against vasculoproliferative disorders.

Methods for identifying cardiovascular agents: a review.

Basic and clinical investigation into many of the diverse aspects of cardiovascular drug discovery employs varied approaches aimed at determining physiologic and pathophysiologic efficacy of candidate agents for therapeutic utility with the ultimate hope of identifying those agents capable of exerting salutary influence upon cardiac and vascular tissues. Promising compounds may then be used for prophylactic cardiovascular protection and for the treatment of various disorders including hypertension, cardiomyopathy, occlusive vascular disease, and heart failure. The invention disclosed in Methods for identifying cardiovascular agents [1] provides screening methods which can be used to identify certain suspected cardiovascular agents that inhibit vascular smooth muscle cell (VSMC) activation and/or proliferation, functional adaptations inherent in the responses to disease or injury, or those that enhance vascular endothelial cell (VEC) activation and/or proliferation, processes thought to provide protection to jeopardized blood vessels. Additionally, these screening assays include agents that activate estrogen responsive genes in vascular cells, considering that estrogen signaling is generally suggested to serve pivotal functions in preventing many of the pathologic mechanisms contributing to occlusive vascular complications. The findings of this primary patent are directly relevant for discoveries in related inventions that disclose various provisions for cardiovascular drug discovery. This review will provide detailed synopses of these function-based screening methods capable of identifying cardiovascular protective agents for use in basic science research and clinical drug discovery.

Salutary properties of YC-1 in the cardiovascular and hematological systems.

The soluble guanylate cyclase (sGC)/cyclic guanosine monophosphate (cGMP) second messenger system provides a complex and highly regulated mechanism for signal transduction events and ensuing functional responses through a cascade of serine/threonine protein kinase-dependent pathways. Nitric oxide (NO) and carbon monoxide (CO), two unique diatomic gases endogenously produced by the respective enzymes nitric oxide synthase (NOS) and heme oxygenase (HO), stimulate cellular sGC and synthesize cGMP within the vasculature. Emerging evidence suggests that the independent NOS and HO systems provide reciprocal and complimentary approaches that act to regulate cardiovascular and hematological homeostasis as well as provide protection to the vasculature in response to inimical stimuli or following the onset of vasoproliferative disease. Recent results from our laboratory and others suggest that the newly identified, chemically synthesized benzyl indazole derivative YC-1 is capable of exerting multifunctional and broad-ranging effects in the cardiovascular and hematological systems. YC-1 has been demonstrated to possess redundant biochemical mechanisms that confer significant stimulation upon NO- and CO-regulated, cyclase-dependent events. Ultimately, these acute molecular processes eventuate in YC-1-dependent modulation of platelet and vascular smooth muscle cell (SMC) and endothelial cell (EC) function under both eutrophic and deleterious conditions. Based on accumulating evidence, YC-1 has been suggested to serve as a potential therapeutic adjuvant to be used in interventional medicine, and these results may indicate the existence of an endogenous " YC-1-like" compound that would be the focus of much anticipated investigation. The purpose of this review, therefore, is to provide update information on the mechanisms and physiologic and pathophysiologic roles of the pivotal new multifunctional agent YC-1 in the cardiovascular and hematological systems, and to provide evidence for its potential use as a clinically relevant salutary agent.