Keiko Sonoda

A type 2 ryanodine receptor variant associated with reduced Ca2+ release and short-coupled torsades de pointes ventricular arrhythmia

BACKGROUND Ventricular fibrillation may be caused by premature ventricular contractions (PVCs) whose coupling intervals are <300 ms, a characteristic of the short-coupled variant of torsades de pointes (scTdP). OBJECTIVE The purpose of this study was to analyze the underlying cardiac ryanodine receptor (RyR2) variants in patients with scTdP. METHODS Seven patients with scTdP (mean age 34 ± 12 years; 4 men and 3 women) were enrolled in this study. The RyR2 gene was screened by targeted gene sequencing methods; variant minor allele frequency was confirmed in 3 databases; and the pathogenicity was investigated in silico analysis using multiple tools. The activity of wild-type and mutant RyR2 channels was evaluated by monitoring Ca2+ signals of HEK293 cells with a [3H]ryanodine binding assay. RESULTS The mean coupling interval of PVCs was 282 ± 13 ms. The 12-lead electrocardiogram had no specific findings except PVCs with an extremely short-coupling interval. Genetic analysis revealed 3 novel RyR2 variants and 1 polymorphism, all located in the cytoplasmic region. p.Ser4938Phe was not detected in 3 databases, and in silico analysis indicated its pathogenicity. In functional analysis, p.Ser4938Phe demonstrated loss of function and impaired RyR2 channel Ca2+ release, while 2 other variants, p.Val1024Ile and p.Ala2673Val, had mild gain-of-function effects but were similar to the polymorphism p.Asn1551Ser. CONCLUSION We identified an RyR2 variant associated with reduced Ca2+ release and short-coupled torsades de pointes ventricular arrhythmia. The mechanisms of arrhythmogenesis remain unclear.

High Frequency of Early Repolarization and Brugada-Type Electrocardiograms in Hypercalcemia.

J wave, or early repolarization has recently been associated with an increased risk of lethal arrhythmia and sudden death, both in idiopathic ventricular fibrillation and in the general population. Hypercalcemia is one of the causes of J point and ST segment elevation, but the relationship has not been well studied. The aim of this study was to examine the effects of hypercalcemia on J point elevation.

Mechanical alternans in human idiopathic dilated cardiomyopathy is caused with impaired force-frequency relationship and enhanced poststimulation potentiation.

Mechanical alternans (MA) is frequently observed in patients with heart failure, and is a predictor of cardiac events. However, there have been controversies regarding the conditions and mechanisms of MA. To clarify heart rate-dependent contractile properties related to MA, we performed incremental right atrial pacing in 17 idiopathic dilated cardiomyopathy (DCM) patients and in six control patients. The maximal increase in left ventricular dP/dt during pacing-induced tachycardia was assessed as the force gain in the force–frequency relationship (FG-FFR), and the maximal increase in left ventricular dP/dt of the first post-pacing beats was examined as the force gain in poststimulation potentiation (FG-PSP). As a result, MA was induced in 9 DCM patients (DCM MA(+)) but not in the other 8 DCM patients (DCM MA(−)), and not in any of the control patients. DCM MA(+) had significantly lower FG-FFR (34.7 ± 40.9 vs 159.4 ± 103.9 mmHg/s, P = 0.0091) and higher FG-PSP (500.0 ± 96.8 vs 321.9 ± 94.9 mmHg/s, P = 0.0017), and accordingly a wider gap between FG-PSP and FG-FFR (465.3 ± 119.4 vs 162.5 ± 123.6 mmHg/s, P = 0.0001) than DCM MA(−) patients. These characteristics of DCM MA(+) showed clear contrasts to those of the control patients. In conclusion, MA is caused with an impaired force–frequency relationship despite significant poststimulation potentiation, suggesting that MA reflects ineffective utilization of the potentiated intrinsic force during tachycardia.

Copy number variations of SCN5A in Brugada syndrome

BACKGROUND Loss-of-function mutations in SCN5A are associated in ∼20% of Brugada syndrome (BrS) patients. Copy number variations (CNVs) have been shown to be associated with several inherited arrhythmia syndromes. OBJECTIVE The purpose of this study was to investigate SCN5A CNVs among BrS probands. METHODS The study cohort consisted of 151 BrS probands who were symptomatic or had a family history of BrS, sudden death, syncope, or arrhythmic diseases. We performed sequence analysis of SCN5A by the Sanger method. For detecting CNVs in SCN5A, we performed multiplex ligation-dependent probe amplification analysis of the 151 BrS probands. RESULTS We identified pathogenic SCN5A mutations in 20 probands by the Sanger method. In 140 probands in whom multiplex ligation-dependent probe amplification was successfully performed, 4 probands were found to present different CNVs (deletion in 3 and duplication in 1). Three of them had fatal arrhythmia events; the remaining 1 was asymptomatic but had a family history. Mean age at diagnosis was 23 ± 14 years. All of the baseline 12-lead electrocardiograms showed PQ-interval prolongation. The characteristics of these 4 probands with CNVs were similar to those of the probands with mutations leading to premature truncation of the protein or missense mutations causing peak INa reduction >90%. CONCLUSION We identified SCN5A CNVs in 2.9% of BrS probands who were symptomatic or had a family history.

Frequency characteristics and associations with the defibrillation threshold of ventricular fibrillation in patients with implantable cardioverter defibrillators.

OBJECTIVE The dominant frequency (DF) in frequency analyses is considered to represent the objective cycle length and complexity of activation under conditions of ventricular fibrillation (VF). However, knowledge regarding the mechanisms determining the DF in human VF is limited. We studied the characteristics of the DF of human VF and relationship between DF and the defibrillation threshold. METHODS Seventy-two implantable cardioverter-defibrillator patients and 211 VF were studied. Using defibrillation tests, we performed a frequency analysis with fast Fourier transformation. The correlations between DF and clinical characteristics, including the defibrillation threshold, were assessed. RESULTS The mean DF of all induced VFs was 5.2±0.8 Hz. The patients were divided into two groups according to DF: the low-DF (DF <5.2 Hz, n=32) and high-DF (DF ≥5.2 Hz, n=40) groups. The frequency of structural heart disease was significantly higher in the low-DF group. In addition, the QRS duration, QT interval and effective refractory period of the right ventricle (RV-ERP) were significantly longer in the low-DF group. A multivariate analysis showed RV-ERP to be the only independent predictor of DF. Excluding patients receiving group III anti-arrhythmic drugs, which are known to have potent defibrillation threshold effects, the defibrillation threshold was significantly lower in the low-DF group (p=0.026). CONCLUSION We found that the DF of human VF is associated with underlying heart disease, the cardiac function, cardiac conduction, ventricular refractoriness and defibrillation threshold. Our findings may be useful for identifying and managing patients with a high defibrillation threshold.

Genetic Basis of Early Repolarization Syndrome

Among patients with idiopathic ventricular fibrillation (IVF), we experienced significantly higher prevalence of early repolarization (ER), which had been considered as a benign ECG finding in the past. In addition, recurrence of fatal ventricular arrhythmia was associated with the presence of ER. In analogy to ER seen in Brugada syndrome (BS), recently, a novel concept for J-wave syndrome (JWS) has been proposed by Dr. Antzelevitch and Dr. Yan, which includes both BS and ER syndrome (ERS). Indeed, there are several clinical similarities between them regarding (1) presence of familial cases, (2) male predominance, (3) predominant VF occurrence in night, (4) bradycardia-induced augmentation of J-point elevation, (5) reduction or elimination of J-point elevation during exercise or atrial fibrillation, and (6) pharmacological usefulness of isoproterenol and quinidine. From the point of genetic basis, there reported several candidate genes responsible for ERS, most of which are overlapped with those associated with Brugada syndrome. This chapter will mainly focus on the genetic background of ERS and discuss its similarity and difference to that in BS.

Multigenerational Inheritance of Long QT Syndrome Type 2 in a Japanese Family.

Congenital long QT syndrome (LQTS) is an important cause of sudden cardiac death in young people without any other structural disease. Mutations in the genes encoding the cardiac ion channels or associated proteins have been shown to result in ion channel dysfunction and thereby causing LQTS. We investigated a Japanese family with LQTS for four generations, with the female family members showing severe symptoms. We performed genetic tests for LQTS-related genes and identified a heterozygous KCNH2 mutation (p.K638del). In the family, the KCNH2 mutation had a very high multigenerational inheritance, and female genotype positives showed more severe phenotypes.