Identification and functional study of genetic polymorphisms in cyclic nucleotide phosphodiesterase 3A (PDE3A)

• Published in: Annals of human genetics Vol. 85; no. 2; pp. 80 - 91
• Main Authors: Kim, You Ran, Yi, MyeongJin, Cho, Sun‐Ah, Kim, Woo‐Young, Min, JungKi, Shin, Jae‐Gook, Lee, Su‐Jun
• Format: Journal Article
• Language: English
• Published: England Wiley Subscription Services, Inc 01-03-2021
• Subjects: 3',5'-Cyclic-nucleotide phosphodiesterase; adenylyl cyclase; Adult; Blood Platelets - drug effects; Cardiomyocytes; cilostazol; Cilostazol - administration & dosage; Cilostazol - adverse effects; Cyclic AMP; Cyclic AMP - genetics; Cyclic Nucleotide Phosphodiesterases, Type 3 - genetics; DNA sequencing; Dose-Response Relationship, Drug; Enzymatic activity; Enzymes; Exome Sequencing; Female; Gene polymorphism; genetic polymorphisms; Humans; Intracellular; Intracellular signalling; Male; Mutation, Missense - genetics; Myocytes; Myocytes, Cardiac - drug effects; Myocytes, Cardiac - metabolism; Phosphodiesterase; phosphodiesterase 3A (PDE3A); Platelet Activation - drug effects; Platelet Activation - genetics; Platelet Aggregation Inhibitors - administration & dosage; Platelet Aggregation Inhibitors - adverse effects; Platelets; Polymorphism, Genetic - genetics; Proteins; Signal Transduction - drug effects; Single-nucleotide polymorphism; Site-directed mutagenesis; Western blotting; adenylyl cyclase; cilostazol; phosphodiesterase 3A (PDE3A); genetic polymorphisms; single nucleotide polymorphism
• ISSN: 0003-4800; 1469-1809
• DOI: 10.1111/ahg.12411

Phosphodiesterase 3A (PDE3A) is an enzyme that plays an important role in the regulation of cyclic adenosine monophosphate (cAMP)–mediated intracellular signaling in cardiac myocytes and platelets. PDE3A hydrolyzes cAMP, which results in a decrease in intracellular cAMP levels and leads to platelet activation. Whole‐exome sequencing of 50 DNA samples from a healthy Korean population revealed a total of 13 single nucleotide polymorphisms including five missense variants, D12N, Y497C, H504Q, C707R, and A980V. Recombinant proteins for the five variants of PDE3A (and wild‐type protein) were expressed in a FreeStyle 293 expression system with site‐directed mutagenesis. The expression of the recombinant PDE3A proteins was confirmed with Western blotting. Catalytic activity of the PDE3A missense variants and wild‐type enzyme was measured with a PDE‐based assay. Effects of the missense variants on the inhibition of PDE3A activity by cilostazol were also investigated. All variant proteins showed reduced activity (33–53%; p < .0001) compared to the wild‐type protein. In addition, PDE3A activity was inhibited by cilostazol in a dose‐dependent manner and was further suppressed in the missense variants. Specifically, the PDE3A Y497C showed significantly reduced activity, consistent with the predictions of in silico analyses. The present study provides evidence that individuals carrying the PDE3A Y497C variant may have lower enzyme activity for cAMP hydrolysis, which could cause interindividual variation in cAMP‐mediated physiological functions.