Intracardiac Electrophysiology Research Articles

Rare Variant in MRC2 Associated With Familial Supraventricular Tachycardia and Wolff-Parkinson-White Syndrome.

Accessory pathways are a common cause of supraventricular tachycardia (SVT) and can lead to sudden cardiac death in otherwise healthy children and adults when associated with Wolff-Parkinson-White syndrome. The goal of this study was to identify genetic variants within a large family with structurally normal hearts affected by SVT and Wolff-Parkinson-White syndrome and determine causality of the gene deficit in a corresponding mouse model. Whole exome sequencing performed on 2 distant members of a 3-generation family in which multiple members were affected by SVT or Wolff-Parkinson-White pattern (preexcitation) on ECG identified MRC2 as a candidate gene. Serial electrocardiograms, intracardiac electrophysiology studies, echocardiography, optical mapping studies, and histology were performed on both Mrc2 mutant and WT (wild-type) mice. A rare HET (heterozygous) missense variant c.2969A>G;p.Glu990Gly (E990G) in MRC2 was identified as the leading candidate gene variant segregating with the cardiac phenotype following an autosomal-dominant Mendelian trait segregation pattern with variable expressivity. In vivo electrophysiology studies revealed reentrant SVT in E990G mice. Optical mapping studies in E990G mice demonstrated abnormal retrograde conduction, suggesting the presence of an accessory pathway. Histological analysis of E990G mouse hearts showed a disordered ECM (extracellular matrix) in the annulus fibrosus. Finally, Mrc2 knockdown in human cardiac fibroblasts enhanced accelerated cell migration. This study identified a rare nonsynonymous variant in the MRC2 gene in individuals with familial reentrant SVT, Wolff-Parkinson-White ECG pattern, and structurally normal hearts. Furthermore, Mrc2 knock-in mice revealed an increased incidence of reentrant SVT and bypass tract formation in the setting of preserved cardiac structure and function.

Chronic angiotensin-converting enzyme inhibition attenuates frailty and protects against atrial fibrillation in aging mice

BackgroundAging is a major risk factor for atrial fibrillation (AF); however, not all individuals age at the same rate. Frailty, which is a measure of susceptibility to adverse health outcomes, can be quantified with a frailty index (FI). ObjectiveThis study aimed to determine the effects of angiotensin-converting enzyme (ACE) inhibition on AF and atrial remodeling in aging and frail mice. MethodsAging mice were treated with the ACE inhibitor enalapril for 6 months beginning at 16.5 months of age and frailty was quantified. AF susceptibility and atrial structure and function were assessed by intracardiac electrophysiology in anesthetized mice, high-resolution optical mapping in intact atrial preparations, patch clamping in isolated atrial myocytes, and histology and molecular biology in atrial tissues. ResultsEnalapril attenuated frailty in aging mice with larger effects in females. AF susceptibility was increased in aging mice but attenuated by enalapril. AF susceptibility and duration also increased as a function of FI score. P-wave duration was increased and atrial conduction velocity was reduced in aging mice and improved after enalapril treatment. Furthermore, P-wave duration and atrial conduction velocity were strongly correlated with FI score. Atrial action potential upstroke velocity (Vmax) and Na+ current (INa) were reduced whereas atrial fibrosis was increased in aging mice. Action potential Vmax, INa, and fibrosis were improved by enalapril and also correlated with FI scores. ConclusionACE inhibition with enalapril attenuates frailty and reduces AF susceptibility in aging mice by preventing atrial electrical and structural remodeling.

Intracardiac electrophysiology to characterize susceptibility to ventricular arrhythmias in murine models.

Introduction: Sudden cardiac death (SCD) and ventricular fibrillation are rare but severe complications of many cardiovascular diseases and represent a major health issue worldwide. Although the primary causes are often acute or chronic coronary diseases, genetic conditions, such as inherited channelopathies or non-ischemic cardiomyopathies are leading causes of SCD among the young. However, relevant experimental models to study the underlying mechanisms of arrhythmias and develop new therapies are still needed. The number of genetically engineered mouse models with cardiac phenotype is growing, making electrophysiological studies in mice essential tools to study arrhythmogenicity and arrhythmia mechanisms and to test novel treatments. Recently, intracardiac catheterization via the jugular vein was described to induce and record ventricular arrhythmias in living anesthetized mice. Several strategies have been reported, developed in healthy wild-type animals and based on aggressive right ventricular stimulation. Methods: Here, we report a protocol based on programmed electrical stimulation (PES) performed in clinical practice in patients with cardiac rhythm disorders, adapted to two transgenic mice models of arrhythmia - Brugada syndrome and cardiolaminopathy. Results: We show that this progressive protocol, based on a limited number of right ventricular extrastimuli, enables to reveal different rhythmic phenotypes between control and diseased mice. In this study, we provide detailed information on PES in mice, including catheter positioning, stimulation protocols, intracardiac and surface ECG interpretation and we reveal a higher susceptibility of two mouse lines to experience triggered ventricular arrhythmias, when compared to control mice. Discussion: Overall, this technique allows to characterize arrhythmias and provides results in phenotyping 2 arrhythmogenic-disease murine models.

Natriuretic Peptide Receptor B Protects Against Atrial Fibrillation by Controlling Atrial cAMP Via Phosphodiesterase 2.

β-AR (β-adrenergic receptor) stimulation regulates atrial electrophysiology and Ca2+ homeostasis via cAMP-dependent mechanisms; however, enhanced β-AR signaling can promote atrial fibrillation (AF). CNP (C-type natriuretic peptide) can also regulate atrial electrophysiology through the activation of NPR-B (natriuretic peptide receptor B) and cGMP-dependent signaling. Nevertheless, the role of NPR-B in regulating atrial electrophysiology, Ca2+ homeostasis, and atrial arrhythmogenesis is incompletely understood. Studies were performed using atrial samples from human patients with AF or sinus rhythm and in wild-type and NPR-B-deficient (NPR-B+/-) mice. Studies were conducted in anesthetized mice by intracardiac electrophysiology, in isolated mouse atrial preparations using high-resolution optical mapping, in isolated mouse and human atrial myocytes using patch-clamping and Ca2+ imaging, and in mouse and human atrial tissues using molecular biology. Atrial NPR-B protein levels were reduced in patients with AF, and NPR-B+/- mice were more susceptible to AF. Atrial cGMP levels and PDE2 (phosphodiesterase 2) activity were reduced in NPR-B+/- mice leading to larger increases in atrial cAMP in the presence of the β-AR agonist isoproterenol. NPR-B+/- mice displayed larger increases in action potential duration and L-type Ca2+ current in the presence of isoproterenol. This resulted in the occurrence of spontaneous sarcoplasmic reticulum Ca2+ release events and delayed afterdepolarizations in NPR-B+/- atrial myocytes. Phosphorylation of the RyR2 (ryanodine receptor) and phospholamban was increased in NPR-B+/- atria in the presence of isoproterenol compared with the wildtypes. C-type natriuretic peptide inhibited isoproterenol-stimulated L-type Ca2+ current through PDE2 in mouse and human atrial myocytes. NPR-B protects against AF by preventing enhanced atrial responses to β-adrenergic receptor agonists.

BS-452682-4 EFFECTS OF CHRONIC ANGIOTENSIN CONVERTING ENZYME INHIBITION ON ATRIAL FUNCTION AND ARRHYTHMOGENESIS IN AGED AND FRAIL MICE

Aging is associated atrial remodeling and increased susceptibility to atrial fibrillation (AF); however, aging-dependent changes can be variable. Frailty measures health status independent of chronological age, can be quantified using a frailty index (FI) and can account for heterogeneity in aging individuals. Angiotensin converting enzyme inhibition (ACEi) can impact frailty however its effects on AF and atrial remodeling are unknown. To determine the effects of chronic ACEi on atrial structure, function, and arrhythmogenesis in aged and frail mice. Wildtype mice were treated with the ACEi enalapril (20 mg/kg/day) or placebo for 6 months starting at 75 weeks of age. Mice aged 10-15 weeks were used as young controls. Mice were used in intracardiac electrophysiology, high resolution optical mapping, patch-clamping, and histology. ACEi reduced FI in aged mice (0.24 vs 0.36; n=27-31; P<0.05) without altering blood pressure. AF susceptibility was increased in aged vs young mice (100 vs 17%; n=6; P<0.05) and reduced in ACEi vs aged mice (43 vs 100%; n=6-7; P<0.05). P wave duration was reduced in ACEi vs aged mice (27.4 vs 32.5 ms; n=7; P<0.05), showed variability within age/treatment groups, and was correlated and graded by FI (R=0.91; n=20; P<0.05). Optical mapping revealed an age dependent reduction in atrial conduction velocity (CV; 35.3 vs 40.2 cm/s; n=6-9; P<0.05) while ACEi improved CV in aged mice (37.7 vs 35.3 cm/s; n=6-9; P=0.05). CV was also correlated with FI score (R=0.72; n=24; P<0.05). Patch-clamp studies in isolated left atrial (LA) myocytes identified a reduction in action potential upstroke velocity (Vmax; 150.8 vs 165.4 V/s; n=18-32; P<0.05) that was associated with a reduction in peak sodium current (INa; -38.3 vs -52.1 pA/pF; n=7-19; P<0.05) in aged vs young mice. INa was increased in ACEi vs aged (-47.2 vs -38.3 pA/pF; n=14–19; P=0.09) and not different (P=0.62) vs young mice. Importantly, Vmax (R=0.32; n=73; P<0.05) and INa (R=0.5; n=36; P < 0.05) were graded by FI. LA fibrosis was increased in aged mice vs young (10.8 vs 3.9%; n=5-8; P<0.05), reduced in ACEi vs aged mice (8.9 vs 10.8%; n=5-8; P<0.05), and strongly correlated with FI (R=0.89; n=19; P<0.05). ACEi reduced frailty in aging mice and attenuated age-related AF and atrial remodeling. Aging was associated with substantial variability within groups. Frailty was a strong predictor of changes atrial function in aging mice including after ACEi treatment, independent of chronological age.

MP-453085-1 CALORIC RESTRICTION ATTENUATES SINOATRIAL NODE AND ATRIAL ARRHYTHMOGENESIS IN AGED AND FRAIL MICE

Atrial fibrillation (AF) and sinoatrial node (SAN) dysfunction are highly prevalent in elderly individuals; however, aging is also highly heterogenous. This heterogeneity has led to the concept of ‘frailty’, a state of increased vulnerability to adverse health outcomes in aging. Mild caloric restriction (CR; reducing caloric intake without malnutrition) can preserve cardiac function and improve frailty (healthspan); however, the effects of late-onset CR on SAN/atrial function in aging mice are unknown. To determine if CR affects frailty and SAN/atrial structure, function and arrhythmogenesis in aging mice. At 56 weeks of age, C57BL/6 mice were placed on either 25% CR or control diet ad libitum. Frailty scores were measured every 4 weeks for 24 weeks, at which point atrial and SAN function was studied using intracardiac electrophysiology, optical mapping, and histology. Young control mice (12 weeks) were used to account for differences in aging. Following 24 weeks of 25% CR, mice exhibited reduced frailty scores compared to the aged control diet (0.207±0.006 vs. 0.273±0.007, n=17-23; P<0.0001). Aged CR mice demonstrated lower AF inducibility (5/16, 31% vs. 12/18, 67%; P<0.05) compared to aged controls, as well as reduced corrected SAN recovery times (27.3±4.5 vs. 42.2±7.1, n=8-15; P<0.05). SAN recovery times in aged CR mice were similar to young mice (27.3±4.5 vs. 24.2.3±6.9, n=13-15; P>0.05). Due to substantial heterogeneity within groups, no difference in P-wave duration (PWD) was observed between groups. However, PWD (R2=0.15, P<0.01) and SAN recovery time (R2=0.19, P<0.0001) were each correlated with FI such that these fell along a continuum regardless of age or diet. Atrial conduction velocities were increased in CR mice compared to aged controls (51.1±2.4 vs. 19.2±2.7, n=5-7; P<0.0001), comparable to young controls (51.1±2.4 vs. 46.3±1.8, n=6-7; P>0.05). Atrial interstitial fibrosis was decreased in aged CR mice compared to aged controls (7.9±0.6 vs. 13.0±2.1, n=3; P<0.05), again comparable to young mice (7.9±0.6 vs. 5.0±0.2, n=3; P>0.05). Both conduction velocity (R2=0.32; P<0.01) and atrial fibrosis (R2=0.81; P<0.0001) showed a strong correlation to frailty score regardless of age or diet. Late-onset CR reduces frailty, improves SAN and atrial structure/function, and attenuates arrhythmogenesis in aging mice. These changes were strongly correlated, and often better predicted, by frailty compared to chronological age.

PO-03-007 CROSS VALIDATION OF A MACHINE LEARNING ALGORITHM DESIGNED TO IDENTIFY PULMONARY VEIN SIGNALS USING COMPETING TRAINING DATA

State of the art electrophysiology (EP) recording systems allow recording of both the surface electrocardiograms and intracardiac electrograms (EGMs) in an unfiltered fashion with improved quality, lower noise and higher sample rate. Legacy EP recording systems sample and store irreversibly pre-filtered EGMs, often susceptible to noise and interference. This study hypothesized that EGM quality impacts the performance of machine learning algorithms later trained on these data. This study compares the performance of a previously validated machine learning algorithm that classifies pulmonary vein potentials trained on simultaneous recorded legacy and novel EP recording system datasets. Data from pulmonary vein isolation procedures were simultaneously collected using a conventional EP recording system (EP-Tracer, CardioTek and LabSystem Pro, Boston Scientific) and a novel electrophysiology recording system (ECGenius System, CathVision, Denmark) connected in parallel. A machine learning classification algorithm, originally designed for legacy data, was adapted to use data from both the conventional and ECGenius EP recording systems. The performance of the algorithm was evaluated using k-fold validation without overlap in patient data between folds. Classified labels across all folds were summarized and compared to the common ground truth labels of the two datasets. In total, 1536 samples from 521 EGMs were extracted from 89 patients prospectively enrolled in the PVISION study. The trained algorithm using the ECGenius recorded EGMs resulted in a higher predictive value (91% vs 84%), sensitivity (93% vs 90%) and overall accuracy (91% vs 86%) compared to the algorithm trained with EGMs from the conventional dataset, resulting in superiority of the ECGenius System algorithm (p <0.05). Receiver operating characteristic (ROC) curves show superiority of the ECGenius-trained algorithm at all relevant operating points (Fig. 1). Performance of machine learning algorithms using intracardiac EP data are dependent on EGM quality and sample rate. Using high quality, unfiltered unipolar EGMs from a novel EP recording system resulted in superior machine learning algorithm performance compared to the use of EGMs recorded with legacy EP systems.

Reduction in Junctophilin 2 Expression in Cardiac Nodal Tissue Results in Intracellular Calcium-Driven Increase in Nodal Cell Automaticity.

Spontaneously depolarizing nodal cells comprise the pacemaker of the heart. Intracellular calcium (Ca2+) plays a critical role in mediating nodal cell automaticity and understanding this so-called Ca2+ clock is critical to understanding nodal arrhythmias. We previously demonstrated a role for Jph2 (junctophilin 2) in regulating Ca2+-signaling through inhibition of RyR2 (ryanodine receptor 2) Ca2+ leak in cardiac myocytes; however, its role in pacemaker function and nodal arrhythmias remains unknown. We sought to determine whether nodal Jph2 expression silencing causes increased sinoatrial and atrioventricular nodal cell automaticity due to aberrant RyR2 Ca2+ leak. A tamoxifen-inducible, nodal tissue-specific, knockdown mouse of Jph2 was achieved using a Cre-recombinase-triggered short RNA hairpin directed against Jph2 (Hcn4:shJph2). In vivo cardiac rhythm was monitored by surface ECG, implantable cardiac telemetry, and intracardiac electrophysiology studies. Intracellular Ca2+ imaging was performed using confocal-based line scans of isolated nodal cells loaded with fluorescent Ca2+ reporter Cal-520. Whole cell patch clamp was conducted on isolated nodal cells to determine action potential kinetics and sodium-calcium exchanger function. Hcn4:shJph2 mice demonstrated a 40% reduction in nodal Jph2 expression, resting sinus tachycardia, and impaired heart rate response to pharmacologic stress. In vivo intracardiac electrophysiology studies and ex vivo optical mapping demonstrated accelerated junctional rhythm originating from the atrioventricular node. Hcn4:shJph2 nodal cells demonstrated increased and irregular Ca2+ transient generation with increased Ca2+ spark frequency and Ca2+ leak from the sarcoplasmic reticulum. This was associated with increased nodal cell AP firing rate, faster diastolic repolarization rate, and reduced sodium-calcium exchanger activity during repolarized states compared to control. Phenome-wide association studies of the JPH2 locus identified an association with sinoatrial nodal disease and atrioventricular nodal block. Nodal-specific Jph2 knockdown causes increased nodal automaticity through increased Ca2+ leak from intracellular stores. Dysregulated intracellular Ca2+ underlies nodal arrhythmogenesis in this mouse model.

Regional and temporal progression of atrial remodeling in angiotensin II mediated atrial fibrillation

Atrial fibrillation (AF) is associated with electrical and structural remodeling in the atria; however, the regional and temporal progression of atrial remodeling is incompletely understood. The objective of this study was to investigate the regional and temporal progression of atrial remodeling leading to changes in AF susceptibility in angiotensin II (Ang II) mediated hypertension. Mice were infused with Ang II for 3, 10 or 21days. AF susceptibility and atrial electrophysiology were studied in vivo using intracardiac electrophysiology. Right and left atrial myocyte electrophysiology was studied using patch-clamping. Atrial fibrosis was assessed histologically. P wave duration and atrial effective refractory period increased progressively from 3 to 21days of Ang II. AF susceptibility tended to be increased at 10days of Ang II and was elevated at 21days of Ang II. Left, but not right, atrial AP upstroke velocity and Na+ current were reduced at 10 and 21days of Ang II. Left atrial action potential (AP) duration increased progressively from 3 to 21days of Ang II due to reductions in repolarizing K+ current. Right atrial AP prolongation was increased only after 21days of Ang II. Left and right atrial fibrosis developed progressively from 3 to 21days, but increases were larger in the left atrium. In conclusion, Ang II mediated atrial electrical and structural remodeling develop earlier and more extensively in the left atrium compared to the right atrium, providing insight into how atrial remodeling leads to enhanced AF susceptibility in Ang II mediated hypertension.

Feasibility of simultaneous multichannel magnetocardiographic mapping, multiple monophasic action potential recording and laser photocoagulation of isolated and perfused rabbit ventricular myocardium

Abstract Background Multichannel magnetocardiographic mapping (MCG) has been validated as an accurate non-invasive method for 3D localization of arrhythmogenic substrates suitable for ablation treatment and of intracardiac electrophysiology catheters (1,2). High resolution (HR) MCG-guided multiple simultaneous monophasic action potential (multi-MAP) recording and pacing with a single patented amagnetic catheter (AC) detects local arrhythmogenic mechanisms (3). Laser photocoagulation (LP) providing more controlled and homogeneous myocardial lesions (4), may be an alternative to radiofrequency (RF) ablation, without unwanted electromagnetic noise effect during real-time MCG. Purpose This study aimed to experimentally test the feasibility of simultaneous multi-MAP recording and LP guided by real-time MCG. Methods An unshielded 36 channels SQUID system (sensitivity of 30 fT/Hz1/2, at 1 Hz) (CMI, USA) was used to study Langendorff-perfused isolated rabbit heart with MCG (Figure 1A). Four epicardial MAPs were simultaneously recorded (inter MAP distance: 1 mm) with a single multi-MAP AC (bandpass: DC-500 Hz; sampling frequency: 2 kHz) (GE-Prucka Cardiolab). LP was performed with a NdYAG laser (Dornier Medilas 5100) and a catheter system for safe myocardium LP (5). The study was carried out according to NIH guidelines for animal care and use. Results MCG imaging of rabbit ventricular magnetic field distribution and inverse source localization with the magnetic dipole model are shown in Figure 1B. The difference in morphology of local HR ventricular multi-MAP signals was compatible with myocardial anisotropy (Figure 1C). With appropriate cooling of the optic fiber-tissue interface, no epicardial damage was induced by LP, whereas deeper well defined intramural myocardial lesions were appreciable, of different size as a function of the energy delivered (Figure 1D). Multi-MAP recording was not significantly disturbed by LP. Keeping the Medilas unit at least 5 meters away from the MCG sensors, no electromagnetic artifacts were observed during MCG. Conclusion MCG, single catheter multi-MAP recording and well-defined LP lesions of ventricular myocardium were simultaneously feasible on Langendorff-perfused isolated rabbit heart, without major electromagnetic interferences, in an unshielded catheterization laboratory. With appropriate technological developments, safe MCG-guided single-catheter laser ablation of focal arrhythmogenic substrates is foreseen as a possible minimally invasive alternative to current interventional approaches. Funding Acknowledgement Type of funding sources: None.

Sodium-glucose cotransporter-2 inhibitor alleviated atrial remodeling in STZ-induced diabetic rats by targeting TLR4 pathway.

PurposeThe mechanism of sodium–glucose cotransporter-2 inhibitor (SGLT-2i) reducing the incidence of atrial fibrillation remains unclear. We hypothesize that sodium–glucose cotransporter-2 inhibitor alleviated atrial remodeling in STZ-induced diabetic rats by targeting TLR4 pathway.MethodsA total of 42 rats were randomly assigned into three groups: control group (CON group); diabetes group (DM group): diabetes mellitus rats were established by 65 mg/kg streptozotocin (STZ) intraperitoneal injection; and diabetes + dapagliflozin group (DM + DAPA group): diabetic rats were given DAPA gavage administration (DAPA 2mg/kg/d for 4 weeks by gavage administration), 14 rats in each group. Epicardial multiple-lead recording and intracardiac electrophysiology studies were performed to investigate the electrical remodeling in the heart and the atrial fibrillation inducibility in each group. Western blot analysis and real-time PCR were used to determine the protein and mRNA expression of toll-like receptor 4 (TLR4), interleukin receptor-associated kinase 1 (IRAK1), tumor necrosis factor receptor-associated factor 6 (TRAF6), nuclear factor-kappa B (NF-κB), and type I collagen (collagen I).ResultsCompared with rats in CON group, rats in DM group showed marked myocardial fibrosis, ectopic pacing excitement, reduced conduction velocity, decreased cardiac function. TLR4/IRAK1/TRAF6/NF-κB, collagen I proteins expressions and incidence of atrial fibrillation (27.3%) were increased in DM group. Parts of these changes were reversed by treatment of DAPA. Incidence of atrial fibrillation was decreased in DM + DAPA group (2.8%).ConclusionsSGLT-2i dapagliflozin may prevent diabetic rats' atrial remodeling and reduce the inducibility of atrial fibrillation partly by targeting TLR4/IRAK1/TRAF6/NF-κB inflammatory pathway.

Abstract P1088: Polycystin-1 Protects Against Experimental Atrial Fibrillation

Atrial fibrillation (AF) is the most common cardiac arrhythmia with a 1 in 4 lifetime risk. AF impairs cardiac function and increases the risk of stroke and heart attack. Although this is a common and harmful disease, little is known about the molecular mechanisms driving this disease. We have previously demonstrated that the protein polycystin-1 (PC1) plays a role in atrial cardiomyocyte excitation-contraction coupling in ventricular cardiomyocytes. Interestingly, patients with mutations in PC1 encoding gene have increased rates of atrial arrhythmias, including atrial fibrillation. Here, we sought to study the role of PC1 in atrial cardiomyocyte function and atrial fibrillation inducibility. We performed intracardiac electrophysiology with burst pacing in two mouse models with impaired PC1 function: 1) a mouse model harboring R3277C mutation (RC/RC) known to cause autosomal polycystic kidney disease, and 2) cardiomyocyte-specific PC1 KO (CKO). We observed increased rates of atrial fibrillation inducibility in both RC/RC and PC1 CKO mice compared to their respective control littermates (76.9% in RC/RC and 81.8% in CKO vs 20.0% and 11.1% in their respective WT controls). To elucidate molecular mechanisms driving increased susceptibility to atrial fibrillation, we determined whether PC1 affects excitation-contraction coupling in atrial cardiomyocytes. Human induced pluripotent stem cells-derived atrial cardiomyocytes (hiPSC-aCM) were treated with control or PC1 siRNA. Action potentials and intracellular calcium were measured using Fluovolt and Fluo-4, respectively. PC1 ablation decreased action potential duration and calcium transient peak amplitude elicited by electrical stimulation in hiPSC-CM. Decreased calcium transient peaks were also observed in HL-1 atrial cell line treated with siRNA and adult atrial cardiomyocytes isolated from RC/RC mice compared to their control counterparts. Unexpectedly, RNA-sequencing revealed altered DNA Damage Response gene expression. DNA damage has been implicated in the pathogenesis of atrial fibrillation. Therefore, we studied whether tachypacing elicited DNA damage in hiPSC-aCM. We observed increased levels of H2AX phosphorylation (gamma H2AX) in hiPSC-aCM with reduced PC1 expression. Our data show that PC1 mutations increase atrial fibrillation inducibility in multiple mouse models, likely due to impaired DNA damage response and altered action potential and calcium handling.

Cover Image, Volume 33, Issue 3

The cover image is based on the Original Article Differentiating left bundle branch pacing and left ventricular septal pacing: An algorithm based on intracardiac electrophysiology by Xing Chen et al., https://doi.org/10.1111/jce.15350.

Differentiating left bundle branch pacing and left ventricular septal pacing: An algorithm based on intracardiac electrophysiology.

Left bundle branch pacing (LBBP) is a new near-physiological pacing modality. Distinguishing left ventricular septal only pacing (LVSP) from nonselective LBBP still needs clarification. This prospective study sought to establish a differentiation algorithm to confirm LBBP. LBBP was attempted in consecutive patients. If direct LBB capture (LBBP) could not be confirmed, LVSP was considered to have been achieved. Intracardiac left ventricular (LV) activation sequence and activation time were analyzed using coronary sinus (CS) electrogram mapping. Electrophysiological parameters including S-CSmax, S-CSmin, LV lateral wall activation time, ΔLV, and LBB potential were compared between LBBP and LVSP. Stimulated LV activation time (S-LVAT) and stimulated QRS duration (S-QRSd) were also compared between the two groups. Multivariate logistic regression analysis was used to develop a prediction algorithm for LBBP. Of the 43 prospectively enrolled patients, 27 underwent LBBP and 16 underwent LVSP. All LBBP patients showed identical LV activation sequences to their intrinsic rhythm while no LVSP patients maintained their intrinsic sequence. S-CSmax, ΔLV, LV lateral wall activation time, and S-LVAT during LBBP were significantly shorter than those during LVSP. Combining LBB potential with S-LVAT had the largest area under the curve (AUC) of 0.985 for confirming LBBP with a sensitivity of 95.2% and a specificity of 93.7%. Compared with LVSP, LBBP preserves a normal LV activation sequence and better electrical synchrony. A combination of LBB potential with S-LVAT can be an effective and practical model to distinguish LBBP from LVSP during implantation in patients with normal LBB activation.

564 Controversial role of intracardiac eletrophysiology study in Brugada syndrome: analysis of a single-centre retrospective cohort study

Abstract Aims According to European guidelines, aborted sudden cardiac death (SCD) in Brugada syndrome (BrS) is regarded as a I class recommendation for secondary prevention implantable cardioverter defibrillator (ICD). However, the risk stratification of BrS patients for primary prevention ICD still represents a clinical conundrum. Although intracardiac electrophysiology (EP) study proved useful for the selection of high-risk patients in this setting. Therefore, aim of this study was to assess all clinical and EP variables associated with the induction of VA at EP study and the rate of appropriate/inappropriate ICD interventions and/or clinical SCD events in these patients occurring at follow-up. Methods and results From 2001 to 2021, all EP studies performed in symptomatic/asymptomatic patients (46 ± 14 years, M 88%) with/without family history of SCD spontaneous/drug-induced type I pattern (TIP) on ECG and no spontaneous ventricular arrhythmias were retrospectively considered at our study centre. Clinical variables, BrS pattern, EP study data (including right ventricular site and type of stimulation protocol), and ICD interventions (DC-shocks or Anti-Tachycardia Pacing events, ATP) and/or SCD events occurring at follow-up were all evaluated. EP study was deemed positive for any polymorphic VA induced during programmed ventricular stimulation; non-sustained episodes included. ICD was routinely implanted in all patients with a positive EP study. Follow-up data were detected by the collection of medical and home-monitoring recordings at study-site level. Follow-up data were available in 50 patients (9 ± 6 years on average). Patients were generally young with few cardiovascular comorbidities. SCD history was known in 21 (42%) with a significant number of asymptomatic patients (48%). Br patterns were equally distributed in the investigated population (spontaneous and drug-induced TIP in 52% and 48%, respectively) and AF history was fairly common (16%). In the study population, EP study tested positive in 30 patients (60%): spontaneous TIP (P = 0.0518), few extrastimuli during programmed ventricular stimulation (P = 0.0015), and right ventricular stimulation at the apical site (P ≤ 0.0001) were the only variables to be clearly associated with a positive EP study in the appraised patients. At follow-up, appropriate ICD shocks were documented in 4 out of 30 implanted patients (13%) at generally 5 ± 7 years from EP study evaluation. Although three ICD interventions (75%) occurred in patients with spontaneous TIP, one patient with drug-induced TIP pattern and positive EP study referred to Emergency Department for unrelenting VT storm after roughly 13 years from ICD implantation. Inappropriate ICD interventions for fast rate AF were detected in 10% of cases. Finally, no SCD events were documented at follow up in patients with a negative EP study. Conclusions In a retrospective analysis, EP study proved useful in the risk stratification of SCD in BrS patients. A few ventricular extrastimuli delivered at the right ventricular apex seem sufficient to prompt the induction of life-threatening VA in high-risk BrS patients during EP study. Moreover, in this setting, a negative EP study seems protective against the development of VA/SCD events at follow-up. However, not only is spontaneous TIP associated with an increased risk of arrhythmic death, but a drug-induced TIP, generally regarded as a low-risk condition, might also be associated with a long-term hazard of SCD in these patients.

Loss of Natriuretic Peptide Receptor C Enhances Sinoatrial Node Dysfunction in Aging and Frail Mice.

Heart rate (HR) is controlled by the sinoatrial node (SAN). SAN dysfunction is highly prevalent in aging; however, not all individuals age at the same rate. Rather, health status during aging is affected by frailty. Natriuretic peptides regulate SAN function in part by activating natriuretic peptide receptor C (NPR-C). The impacts of NPR-C on HR and SAN function in aging and as a function of frailty are unknown. Frailty was measured in aging wild-type and NPR-C knockout (NPR-C-/-) mice using a mouse clinical frailty index (FI). HR and SAN structure and function were investigated using intracardiac electrophysiology in anesthetized mice, high-resolution optical mapping in intact atrial preparations, histology, and molecular biology. NPR-C-/- mice rapidly became frail leading to shortened life span. HR was reduced and SAN recovery time was increased in older versus younger mice, and these changes were exacerbated in NPR-C-/- mice; however, there was substantial variability among age groups and genotypes. HR and SAN recovery time were correlated with FI score and fell along a continuum regardless of age or genotype. Optical mapping demonstrates impairments in SAN function that were also correlated with FI score. SAN fibrosis was increased in aged and NPR-C-/- mice and was graded by FI score. Loss of NPR-C results in accelerated aging and rapid decline in health status in association with impairments in HR and SAN function. Frailty assessment was effective and better able to distinguish aging-dependent changes in SAN function in the setting of shortened life span due to loss of NPR-C.

Distinct Effects of Ibrutinib and Acalabrutinib on Mouse Atrial and Sinoatrial Node Electrophysiology and Arrhythmogenesis.

BackgroundIbrutinib and acalabrutinib are Bruton tyrosine kinase inhibitors used in the treatment of B‐cell lymphoproliferative disorders. Ibrutinib is associated with new‐onset atrial fibrillation. Cases of sinus bradycardia and sinus arrest have also been reported following ibrutinib treatment. Conversely, acalabrutinib is less arrhythmogenic. The basis for these different effects is unclear.Methods and ResultsThe effects of ibrutinib and acalabrutinib on atrial electrophysiology were investigated in anesthetized mice using intracardiac electrophysiology, in isolated atrial preparations using high‐resolution optical mapping, and in isolated atrial and sinoatrial node (SAN) myocytes using patch‐clamping. Acute delivery of acalabrutinib did not affect atrial fibrillation susceptibility or other measures of atrial electrophysiology in mice in vivo. Optical mapping demonstrates that ibrutinib dose‐dependently impaired atrial and SAN conduction and slowed beating rate. Acalabrutinib had no effect on atrial and SAN conduction or beating rate. In isolated atrial myocytes, ibrutinib reduced action potential upstroke velocity and Na+ current. In contrast, acalabrutinib had no effects on atrial myocyte upstroke velocity or Na+ current. Both drugs increased action potential duration, but these effects were smaller for acalabrutinib compared with ibrutinib and occurred by different mechanisms. In SAN myocytes, ibrutinib impaired spontaneous action potential firing by inhibiting the delayed rectifier K+ current, while acalabrutinib had no effects on SAN myocyte action potential firing.ConclusionsIbrutinib and acalabrutinib have distinct effects on atrial electrophysiology and ion channel function that provide insight into the basis for increased atrial fibrillation susceptibility and SAN dysfunction with ibrutinib, but not with acalabrutinib.

Dyssynchronous cardiomyopathy in patients with WPW syndrome and its reversibility after RFA of accessory pathway

Abstract Introduction In some cases accessory pathway-mediated ventricular preexcitation can be associated with electro-mechanical dyssynchrony and, consequently, with dyssynchrony-related dilated cardiomyopathy, even in the absence of supraventricular tachycardia (SVT). Sometimes rapid progression of ventricular dysfunction developed in such patients after birth. Methods and materials 8 patients with asymptomatic WPW and dyssynchronous cardiomyopathy were examined in our Institute from 2017 till 2019. Four patients of the group were observed in other clinics with dilated cardiomyopathy (DCM) and prescribed appropriate therapy without significant clinical effect. The absence of complaints of heartbeat and episodes of tachycardia at the scheduled Holter monitoring allowed eliminating incessant tachycardia as a possible cause of cardiomyopathy. After radiofrequency ablation (RFA) all patients were performed ECG to assess QRS duration, Holter monitoring, echocardiography (Echo) for assessment of heart chamber volume and left ventricle (LV) contractile function and Speckle tracking – Echo with LV longitudinal strain assessment. The average age of the patients was 9.5 years (from 2 to 14 years). 6 children had heart failure (NYHA Class II). Results According to Echo all patients had widened QRS complex. Dilatation and enlargement of LV volume were marked in 6 pts. According to Echo7 patients had decrease of LV contractile function. According to Speckle tracking – Echo all patients had intraventricular dyssynchrony, decrease of longitudinal strain indices. At intracardiac electrophysiology study right-sided accessory pathways was diagnosed in all patients, successful RFA was performed. Preexitation relapsed, and the repeated RFA was performed in 2 pts. Complications caused by RFA were not marked. After RFA all patients showed a regular normalization of QRS duration. At the 5th day after RFA Echo was performed to all patients. The patients with initially decreased LV ejection fraction had it increased. The patients with initial dilatation and LV volume enlargement had normalization of the given indices. According to Speckle tracking – Echo normalization of global and regional myocardial function, LV longitudinal strain, disappearance of intraventricular and interventricular dyssynchrony were marked in all patients. The index of longitudinal strain was 17,0±0,64% before RFA, after RFA - 23,75±0,92% (p=0,ehab724.035946). Conclusion As a result of RFA of accessory pathways electromechanical resynchronization causes LV demodeling and restoration of its contractile ability. The presented clinical examples are an additional indication for RFA of accessory pathways even in the absence of SVT in patients regardless of age. Funding Acknowledgement Type of funding sources: None.

Atrial Fibrillation in Aging and Frail Mice: Modulation by Natriuretic Peptide Receptor C.

[Figure: see text].

Natriuretic peptide receptor B maintains heart rate and sinoatrial node function via cyclic GMP-mediated signalling.

Heart rate (HR) is a critical indicator of cardiac performance that is determined by sinoatrial node (SAN) function and regulation. Natriuretic peptides, including C-type NP (CNP), have been shown to modulate ion channel function in the SAN when applied exogenously. CNP is the only NP that acts as a ligand for natriuretic peptide receptor-B (NPR-B). Despite these properties, the ability of CNP and NPR-B to regulate HR and intrinsic SAN automaticity in vivo, and the mechanisms by which it does so, are incompletely understood. Thus, the objective of this study was to determine the role of NPR-B signalling in regulating HR and SAN function. We have used NPR-B deficient mice (NPR-B+/-) to study HR regulation and SAN function using telemetry in conscious mice, intracardiac electrophysiology in anaesthetized mice, high-resolution optical mapping in isolated SAN preparations, patch-clamping in isolated SAN myocytes, and molecular biology in isolated SAN tissue. These studies demonstrate that NPR-B+/- mice exhibit slow HR, increased corrected SAN recovery time, and slowed SAN conduction. Spontaneous AP firing frequency in isolated SAN myocytes was impaired in NPR-B+/- mice due to reductions in the hyperpolarization activated current (If) and L-type Ca2+ current (ICa,L). If and ICa,L were reduced due to lower cGMP levels and increased hydrolysis of cAMP by phosphodiesterase 3 (PDE3) in the SAN. Inhibiting PDE3 or restoring cGMP signalling via application of 8-Br-cGMP abolished the reductions in cAMP, AP firing, If, and ICa,L, and normalized SAN conduction, in the SAN in NPR-B+/- mice. NPR-B+/- mice did not exhibit changes in SAN fibrosis and showed no evidence of cardiac hypertrophy or changes in ventricular function. NPR-B plays an essential physiological role in maintaining normal HR and SAN function by modulating ion channel function in SAN myocytes via a cGMP/PDE3/cAMP signalling mechanism.