A novel human R25C-phospholamban mutation is associated with super-inhibition of calcium cycling and ventricular arrhythmia

作者：Liu Guan Sheng; Morales Ana; Vafiadaki Elizabeth; Lam Chi Keung; Cai Wen Feng; Haghighi Kobra; Adly George; Hershberger Ray E; Kranias Evangelia G^*

来源：Cardiovascular Research, 2015, 107(1): 164-174.

DOI：10.1093/cvr/cvv127

• Dilated cardiomyopathy
• Calcium cycling
• Mutation

Aims Depressed sarcoplasmic reticulum (SR) Ca2+ cycling, a universal characteristic of human and experimental heart failure, may be associated with genetic alterations in key Ca2+-handling proteins. In this study, we identified a novel PLN mutation (R25C) in dilated cardiomyopathy (DCM) and investigated its functional significance in cardiomyocyte Ca2+-handling and contractility.
Methods and results Exome sequencing identified a C73T substitution in the coding region of PLN in a family with DCM. The four heterozygous family members had implantable cardiac defibrillators, and three developed prominent ventricular arrhythmias. Overexpression of R25C-PLN in adult rat cardiomyocytes significantly suppressed the Ca2+ affinity of SR Ca2+-ATPase (SERCA2a), resulting in decreased SR Ca2+ content, Ca2+ transients, and impaired contractile function, compared with WT-PLN. These inhibitory effects were associated with enhanced interaction of R25C-PLN with SERCA2, which was prevented by PKA phosphorylation. Accordingly, isoproterenol stimulation relieved the depressive effects of R25C-PLN in cardiomyocytes. However, R25C-PLN also elicited increases in the frequency of Ca2+ sparks and waves as well as stress-induced aftercontractions. This was accompanied by increased Ca2+/calmodulin-dependent protein kinase II activity and hyper-phosphorylation of RyR2 at serine 2814.
Conclusion The findings demonstrate that human R25C-PLN is associated with super-inhibition of SERCA2a and Ca2+ transport as well as increased SR Ca2+ leak, promoting arrhythmogenesis under stress conditions. This is the first mechanistic evidence that increased PLN inhibition may impact both SR Ca2+ uptake and Ca2+ release activities and suggests that the human R25C-PLN may be a prognostic factor for increased ventricular arrhythmia risk in DCM carriers.

• 出版日期2015-7-1