(M1030-04-24) Old Dog Learning a New Trick: Statins as Novel Inhibitors of Vascular PDE1A for Regulating Vasodilation and Blood Pressure

Purpose: Statins are the world's most prescribed drugs, used by over 200 million people. Studies over the past three decades suggest that long-term statin use produces numerous beneficial cardiovascular effects mediated by either cholesterol-dependent or -independent mechanisms, involving Rho kinase (ROCK) signaling. Using a combination of artificial intelligence (AI) analysis followed by extensive wet laboratory testing, we unveil a novel role for commonly used statins as selective inhibitors of phosphodiesterase 1A (PDE1A), inducing rapid vasodilation and blood pressure reduction.

Methods: We conducted unbiased in silico blind docking analysis to determine the interactibility of statins with PDE1A, a major cGMP-hydrolyzing enzyme in smooth muscle cells. To further validate this experimentally, we performed in vitro PDE activity assays using purified human PDE1A and PDE5A. After establishing that statins are selective inhibitors of PDE1A, we tested the significance of such action in resistance mesenteric arteries. We used pressurized arterial myography to track the diameter of murine resistance mesenteric arteries. Isolated arterial segments were cannulated and maintained at 37°C in a perfusion chamber. Arteries were allowed to develop myogenic tone at 80 mmHg, and statins (0.01-10 nM) and pharmacological modulators were applied. To determine whether PDE1A inhibition and vasodilation could regulate blood pressure, we used tail cuff analysis of blood pressure in healthy, awake mice. To further understand the role of PDE1A, we generated PDEA1^+/- mice from cryopreserved sperm containing an in-frame deletion of 15 bp that encodes residues within the active site Zn²⁺-binding site of PDE1A, and validated them using polymerase chain reaction (PCR), Western blotting, and PDE activity assays. Complementary biochemical measurements, such as cGMP analysis, were performed according to the manufacturer's instructions.

Results: Using unbiased in silico blind docking, we found that simvastatin, rosuvastatin, and atorvastatin reproducibly dock to the catalytic pocket of both rat and human PDE1A. Our in vitro PDE activity assays demonstrate that while all statins dose-dependently inhibit PDE1A, none of the statins inhibit the activity of PDE5A. Consistent with this, our pressure myography data show that rosuvastatin application (0.01-100 nM) induces rapid vasodilation in WT mice mesenteric arteries. In contrast, genetic deletion of PDE1A significantly blunted rosuvastatin-evoked vasodilation from baseline, demonstrating that PDE1A is a molecular target for rapid statin-induced vasodilation. The development of myogenic tone at 20-100 mmHg was significantly attenuated in PDE1A^+/- arteries, which may explain the blunted response in the PDE1A^+/- arteries. Our BP data reveal that the mean arterial pressure (MAP) in saline-treated WT and PDE1A^+/- mice is 101 and 83 mmHg, respectively, demonstrating a significant (p < 0.05) BP reduction in the latter due to PDE1A knockdown. Administration of rosuvastatin and simvastatin in WT mice produces a dose-dependent BP reduction, which was blunted in PDE1A^+/- mice, suggesting a PDE1A-dependent BP reduction mechanism. Interestingly, although the baseline BP is lower in the PDE1A^+/- mice, statin treatment produces further reduction of BP. This is indicative of inhibition of the residual PDE1A by statins, leading to BP reduction, which is consistent with the inhibition of the remaining PDE1A activity in PDE1A^+/- mice arteries.

Conclusion: Statins are novel selective inhibitors of PDE1A in the vasculature. Direct inhibition of PDE1A underlies statin-induced rapid vasodilation and blood pressure reduction in healthy animals with intact nitric oxide (NO) signaling. Overall, this study unveils a novel role for statins in regulating arterial contractility and blood pressure via a non-canonical pathway involving PDE1A.

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Acknowledgements: This work was funded by NIH R15 REAP Research grant to Dr. Raquibul Hasan as well as an AIREA Award by American Heart Association to Dr. Raquibul Hasan


Fig. 1. (A) Mechanism of interactions between statins and active site of human PDE1A. (B-F): 2D ligand interaction diagram for IBMX (B), ITI-214 (C), and three statins (D-F).


Fig. 2. PDE1A and PDE5A expression in PDE1A+/- mice mesenteric arteries (MAs) and activity in aortas. (A-B) Western blot images for PDE1A and PDE5A expression in MAs (A) and Mean data (B), n=4 * P<0.05 vs WT. (C-D) Mean data for PDE1A (C) and PDE5A (D) activity, n=4 * P<0.05 vs WT. T-test. (E-F) Statins lower BP in a PDE1A-dependent manner. Reduction of BP by rosuvastatin (A) and simvastatin (B). n = 8. * P<0.05 vs Saline, Ŧ P<0.05 vs WT. One-way ANOVA with Tukey


Fig. 3: Statin-induced vasodilation is attenuated in PDE1A+/- mice RMAs. (A) Original traces for rosuvastatin-elicited vasodilation in WT and PDE1A+/- RMAs, and (B) Mean data. n=5, * P<0.05 vs WT. T-test. (C-D): Genetic deletion attenuates myogenic tone in PDE1A+/- mice RMAs and elevates [cGMP]i in aorta. (A) Myogenic tone in RMAs and (B) [cGMP] in WT and PDE1A+/- mice aortas. n=5, * P<0.05 vs WT. T-test.