Gil J. Gross

Biowire: a platform for maturation of human pluripotent stem cell–derived cardiomyocytes

Directed differentiation protocols enable derivation of cardiomyocytes from human pluripotent stem cells (hPSCs) and permit engineering of human myocardium in vitro. However, hPSC-derived cardiomyocytes are reflective of very early human development, limiting their utility in the generation of in vitro models of mature myocardium. Here we describe a platform that combines three-dimensional cell cultivation with electrical stimulation to mature hPSC-derived cardiac tissues. We used quantitative structural, molecular and electrophysiological analyses to explain the responses of immature human myocardium to electrical stimulation and pacing. We demonstrated that the engineered platform allows for the generation of three-dimensional, aligned cardiac tissues (biowires) with frequent striations. Biowires submitted to electrical stimulation had markedly increased myofibril ultrastructural organization, elevated conduction velocity and improved both electrophysiological and Ca2+ handling properties compared to nonstimulated controls. These changes were in agreement with cardiomyocyte maturation and were dependent on the stimulation rate.

The Homeodomain Transcription Factor Irx5 Establishes the Mouse Cardiac Ventricular Repolarization Gradient

Rhythmic cardiac contractions depend on the organized propagation of depolarizing and repolarizing wavefronts. Repolarization is spatially heterogeneous and depends largely on gradients of potassium currents. Gradient disruption in heart disease may underlie susceptibility to fatal arrhythmias, but it is not known how this gradient is established. We show that, in mice lacking the homeodomain transcription factor Irx5, the cardiac repolarization gradient is abolished due to increased Kv4.2 potassium-channel expression in endocardial myocardium, resulting in a selective increase of the major cardiac repolarization current, I(to,f), and increased susceptibility to arrhythmias. Myocardial Irx5 is expressed in a gradient opposite that of Kv4.2, and Irx5 represses Kv4.2 expression by recruiting mBop, a cardiac transcriptional repressor. Thus, an Irx5 repressor gradient negatively regulates potassium-channel-gene expression in the heart, forming an inverse I(to,f) gradient that ensures coordinated cardiac repolarization while also preventing arrhythmias.

Long QT, syndactyly, joint contractures, stroke and novel CACNA1C mutation: expanding the spectrum of Timothy syndrome.

Timothy syndrome (TS) is an autosomal dominant condition with the constellation of features including prolonged QT interval, hand and foot abnormalities, and mental retardation or autism. Splawski et al. [2004] previously described two phenotypes associated with TS distinguished by two unique and different mutations within the CACNA1C gene. We report on a newborn who presented with prolonged QT interval and associated polymorphic ventricular tachycardia, dysmorphic facial features, syndactyly of the hands and feet, and joint contractures, suggestive of TS. He developed a stroke, subsequent intractable seizures, and was found to have cortical blindness and later profound developmental delay. Initial targeted mutation analysis did not identify either of the previously described TS associated mutations; however, full gene sequencing detected a novel CACNA1C gene mutation (p.Ala1473Gly). The clinical and genetic findings in our case expand both the clinical and molecular knowledge of TS.

Maturational and hemodynamic factors predictive of increased cyanosis after bidirectional cavopulmonary anastomosis

Bidirectional cavopulmonary anastomosis (BCA) is thought to be beneficial in the palliation of patients with univentricular congenital heart disease considered at high risk for Fontan repair. Experience with patients undergoing BCA suggested that those who were older or larger at the time of surgery tended to be more cyanotic postoperatively than their younger and smaller counterparts. This study was designed to identify correlates of systemic arterial oxygen saturation after BCA. Specifically, it was postulated that maturational changes in blood flow distribution might be associated with decreasing arterial oxygen saturation. Database records of all 110 patients undergoing BCA at our institution from June 1988 until the end of 1991 were reviewed. Postoperative catheterization data were available for 66 patients. Twenty-one patients were excluded because they had potentially reversible causes of cyanosis yielding inestimable degrees of error in hemodynamic calculations. In the remaining 45 patients, univariate and multivariate regression analyses were used to identify correlates of systemic arterial oxygen saturation. Growth and maturation as represented by body surface area exhibited a highly significant inverse correlation with arterial oxygen saturation (p = 0.005), as did pulmonary vascular resistance (p = 0.003). Patients who underwent BCA when > 3.9 years of age or with body surface area > 0.65 m2 were at significantly increased risk for excessive postoperative cyanosis, defined as systemic arterial oxygen saturation < or = 75% (p < 0.005). The interval between surgery and catheterization correlated directly with arterial oxygen saturation (p = 0.002), indicating a tendency toward earlier study of more cyanotic patients. None of the other variables examined correlated significantly with arterial oxygen saturation.(ABSTRACT TRUNCATED AT 250 WORDS)

Sinus Bradycardia After Intravenous Pulse Methylprednisolone

High-dose intravenous pulse methylprednisolone is an important therapeutic modality for many autoimmune conditions in both children and adults. Adverse effects of this therapy include hypertension, hyperglycemia, and, in children, behavioral changes. Cardiac rhythm disturbances, both tachyarrhythmias and bradyarrhythmias, have been reported in adults but much less commonly in children. Here we report our experience over a 6-month period with 5 children with rheumatic diseases who developed sinus bradycardia during consecutive daily therapy with intravenous pulse methylprednisolone. Reductions in resting heart rate of between 35% and 50% of baseline were observed in each case. All patients were asymptomatic, and all recovered spontaneously over a variable period of time after cessation of pulse therapy. Sinus bradycardia after repeated administration of high-dose pulse methylprednisolone in children may be more common than previously appreciated.

Characteristics, management, and midterm outcome in infants with atrioventricular nodal reentry tachycardia

Atrioventricular nodal reentry is a commonly recognized mechanism of supraventricular tachycardia (SVT) in adults, but is only rarely documented in the first year of life. The aim of this study was to elucidate characteristics, management, and outcome in infants with atrioventricular nodal reentrant tachycardia (AVNRT). Electrophysiologic studies performed between January 1988 and June 1996 were reviewed. Fifteen infants with AVNRT at 58 +/- 27 days (mean +/- SEM) were identified. Five had AVNRT detected following palliation of structural cardiac anomalies, including 4 with critical obstructions to left ventricular outflow. Typical AVNRT (ventriculoatrial interval 49 +/- 5 ms) was observed in 14 of 15 patients and atypical AVNRT (ventriculoatrial interval 191 +/- 22 ms) in 4 of 15. All patients received long-term therapy, beginning with digoxin in 13. Eight had symptomatic recurrences on digoxin and 6 of these were given beta blockers, with satisfactory control in 4. Three patients were controlled with class III agents, and 2 underwent slow pathway radiofrequency modification at ages 4.1 and 6.7 years, respectively. AVNRT was still inducible in 6 of 6 asymptomatic patients who underwent follow-up atrial stimulation studies after discontinuation of medical therapy. All 15 patients were alive with either absent or well-controlled AVNRT at age 45 +/- 7 months. We conclude that the course and outcome of AVNRT diagnosed in the first year of life are generally benign, but that a minority of patients have symptoms persisting beyond infancy. Digoxin is of questionable benefit in long-term control. AVNRT often remains inducible in asymptomatic patients, although the significance of this finding remains to be determined by long-term follow-up.

Diagnosis-specific characteristics of ventricular tachycardia in children with structurally normal hearts

BACKGROUND Improved mechanistic insights and clinical tools provide increasing diagnostic refinement for ventricular tachycardia in young patients with structurally normal hearts, yielding potentially important prognostic and management implications. OBJECTIVE The purpose of this study was to survey the clinical characteristics and outcomes of otherwise healthy children with ventricular tachycardia (VT) who were classified according to contemporary diagnostic criteria. METHODS A single-center retrospective review of patients younger than 18 years of age with VT diagnosed between January 1980 and December 2005 was undertaken. Patients with significant systemic illness or with underlying structural or functional heart disease were excluded. RESULTS A total of 77 patients met inclusion criteria and were grouped as follows: accelerated idioventricular rhythm (AIVR; n = 19), right ventricular tachycardia (RVT; n = 30), left ventricular tachycardia (LVT; n = 23), and catecholamine sensitive polymorphic VT (CPVT; n = 5). AIVR patients were youngest at diagnosis and had the most benign natural history, while the opposite was true of CPVT patients. Ablation was attempted in 2/30 RVT (50% success) and 10/23 LVT (80% success) patients. Severe cardiac events occurred in 3/23 LVT (no deaths) and 2/5 CPVT (one death) patients. CONCLUSION The natural history and appropriate management of VT in young patients with structurally normal hearts are highly dependent on the specific diagnosis. LVT and CPVT are associated with significantly greater morbidity and mortality than AIVR and RVT.

Integrin-linked kinase mediates force transduction in cardiomyocytes by modulating SERCA2a/PLN function.

Human dilated cardiomyopathy (DCM) manifests as a profound reduction in biventricular cardiac function that typically progresses to death or cardiac transplantation. There is no effective mechanism-based therapy currently available for DCM, in part because the transduction of mechanical load into dynamic changes in cardiac contractility (termed mechanotransduction) remains an incompletely understood process during both normal cardiac function and in disease states. Here we show that the mechanoreceptor protein integrin-linked kinase (ILK) mediates cardiomyocyte force transduction through regulation of the key calcium regulatory protein sarcoplasmic/endoplasmic reticulum Ca(2+)ATPase isoform 2a (SERCA-2a) and phosphorylation of phospholamban (PLN) in the human heart. A non-oncogenic ILK mutation with a synthetic point mutation in the pleckstrin homology-like domain (ILK(R211A)) is shown to enhance global cardiac function through SERCA-2a/PLN. Thus, ILK serves to link mechanoreception to the dynamic modulation of cardiac contractility through a previously undiscovered interaction with the functional SERCA-2a/PLN module that can be exploited to rescue impaired mechanotransduction in DCM.

Utility of Exercise Testing in Children and Teenagers With Arrhythmogenic Right Ventricular Cardiomyopathy

Arrhythmogenic right ventricular cardiomyopathy (ARVC) is increasingly recognized as an important cause of exertional sudden death in otherwise healthy young individuals and athletes. Graded exercise testing is routinely included in the evaluation of patients with suspected ARVC, but its diagnostic utility has not been systematically assessed. Using a single-center retrospective design, the rhythm response to graded exercise testing was examined in 33 tests performed in 16 young (aged <18 years) patients with established diagnosis of ARVC. Ventricular premature complexes (VPCs) were classified as absent (graded 0), as being isolated or in couplets (graded 1), or as comprising nonsustained ventricular tachycardia (graded 2) during pretest rest, at peak exercise, and during postexercise recovery. VPCs were absent at rest in 21 of 33 studies, subsequently appearing at peak exercise in 4 studies and during recovery in 2 studies. Isolated VPCs and couplets were present at rest in 9 of 33 studies, with subsequent exercise provoking higher grade ectopic activity in 2 instances at peak exercise and in 1 case during recovery, while VPCs decreased or remained unchanged in all other cases. In all 3 instances in which ventricular tachycardia was observed during pretest rest, there was either suppression (3 at peak exercise, 2 during recovery) or no change (1 case during recovery) in VPC grade. In conclusion, the exercise response of ventricular ectopic activity is highly variable in young patients with ARVC. The diagnostic utility of graded exercise testing is thus questionable in young patients with suspected ARVC, and the absence or suppression of VPCs during exercise should not be considered reassuring in terms of its diagnostic exclusion.

Mutation Location Effect on Severity of Phenotype During Exercise Testing in Type 1 Long-QT Syndrome: Impact of Transmembrane and C-Loop Location

Targeted mutation site‐specific differences have correlated C‐loop missense mutations with worse outcomes and increased benefit of beta‐blockers in LQT1. This observation has implicated the C‐loop region as being mechanistically important in the altered response to sympathetic stimulation known to put patients with LQT1 at risk of syncope and sudden cardiac death.