Abstract
Catecholaminergic polymorphic ventricular tachycardia (CPVT) is an inherited, lethal ventricular arrhythmia triggered by catecholamines. Mutations in genes that encode cardiac ryanodine receptor (RyR2) and proteins that regulate RyR2 activity cause enhanced diastolic Ca2+ release (leak) through the RyR2 channels, resulting in CPVT. Current therapies for CPVT are limited. We found that Z16b, a meroterpenoid isolated from Ganoderma cochlear, inhibited Ca2+ spark frequency (CaSF) in R2474S/ + cardiomyocytes in a dose-dependent manner, with an IC50 of 3.2 μM. Z16b also dose-dependently suppressed abnormal post-pacing Ca2+ release events. Intraperitoneal injection (i.p.) of epinephrine and caffeine stimulated sustained ventricular tachycardia in all R2474S/+ mice, while pretreatment with Z16b (0.5 mg/kg, i.p.) prevented ventricular arrhythmia in 9 of 10 mice, and Z16b administration immediately after the onset of VT abolished sVT in 9 of 12 mice. Of translational significance, Z16b significantly inhibited CaSF and abnormal Ca2+ release events in human CPVT iPS-CMs. Mechanistically, Z16b interacts with RyR2, enhancing the “zipping” state of the N-terminal and central domains of RyR2. A molecular docking simulation and point mutation and pulldown assays identified Z16b forms hydrogen bonds with Arg626, His1670, and Gln2126 in RyR2 as a triangle shape that anchors the NTD and CD interaction and thus stabilizes RyR2 in a tight “zipping” conformation. Our findings support that Z16b is a novel RyR2 stabilizer that can prevent CPVT. It may also serve as a lead compound with a new scaffold for the design of safer and more efficient drugs for treating CPVT.
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Acknowledgements
We appreciate Dr. Long-sheng Song for critical comments. This study was supported by National Natural Science Foundation of China (Grant No. 81970250, 31671179), Natural Science Foundation of Guangdong, China (Grant No. 2021A1515010787), Shenzhen Key Laboratory of Metabolism and Cardiovascular Homeostasis (Grant No. ZDSYS20190902092903 237), Basic Research Foundation of Shenzhen (Grant No. JCYJ20180508152222104, JCYJ20190808123018993, JCYJ20190808121819024, JCYJ20210324094006018), and Shenzhen Science and Technology Innovation Committee Project (Grant No. JCYJ20180507182239272).
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JL generated the hypotheses and designed the experiments, and YXC provided natural product libraries for drug screening including Z16b. JFW performed most experiments and created the figures. GW participated in the key experiments and created the figures. FYQ and DLH prepared and purified Z16b for this study. YW and ALS carried out the molecular docking simulation. HPZ and CLW performed the MD simulation. SYZ and YL provided the clinical sample.
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Wan, Jf., Wang, G., Qin, Fy. et al. Z16b, a natural compound from Ganoderma cochlear is a novel RyR2 stabilizer preventing catecholaminergic polymorphic ventricular tachycardia. Acta Pharmacol Sin 43, 2340–2350 (2022). https://doi.org/10.1038/s41401-022-00870-1
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DOI: https://doi.org/10.1038/s41401-022-00870-1
