Increased Ca 2+ Transient Underlies RyR2-Related Left Ventricular Noncompaction

Abstract

Background: A loss-of-function cardiac ryanodine receptor (RyR2) mutation, I4855M +/– , has recently been linked to a new cardiac disorder termed RyR2 Ca 2+ release deficiency syndrome (CRDS) as well as left ventricular noncompaction (LVNC). The mechanism by which RyR2 loss-of-function causes CRDS has been extensively studied, but the mechanism underlying RyR2 loss-of-function-associated LVNC is unknown. Here, we determined the impact of a CRDS-LVNC-associated RyR2-I4855M +/– loss-of-function mutation on cardiac structure and function. Methods: We generated a mouse model expressing the CRDS-LVNC-associated RyR2-I4855M +/– mutation. Histological analysis, echocardiography, ECG recording, and intact heart Ca 2+ imaging were performed to characterize the structural and functional consequences of the RyR2-I4855M +/– mutation. Results: As in humans, RyR2-I4855M +/– mice displayed LVNC characterized by cardiac hypertrabeculation and noncompaction. RyR2-I4855M +/– mice were highly susceptible to electrical stimulation–induced ventricular arrhythmias but protected from stress-induced ventricular arrhythmias. Unexpectedly, the RyR2-I4855M +/– mutation increased the peak Ca 2+ transient but did not alter the L-type Ca 2+ current, suggesting an increase in Ca 2+ -induced Ca 2+ release gain. The RyR2-I4855M +/– mutation abolished sarcoplasmic reticulum store overload–induced Ca 2+ release or Ca 2+ leak, elevated sarcoplasmic reticulum Ca 2+ load, prolonged Ca 2+ transient decay, and elevated end-diastolic Ca 2+ level upon rapid pacing. Immunoblotting revealed increased level of phosphorylated CaMKII (Ca 2+ -calmodulin dependent protein kinases II) but unchanged levels of CaMKII, calcineurin, and other Ca 2+ handling proteins in the RyR2-I4855M +/– mutant compared with wild type. Conclusions: The RyR2-I4855M +/– mutant mice represent the first RyR2-associated LVNC animal model that recapitulates the CRDS-LVNC overlapping phenotype in humans. The RyR2-I4855M +/– mutation increases the peak Ca 2+ transient by increasing the Ca 2+ -induced Ca 2+ release gain and the end-diastolic Ca 2+ level by prolonging Ca 2+ transient decay. Our data suggest that the increased peak-systolic and end-diastolic Ca 2+ levels may underlie RyR2-associated LVNC.