Unidirectional Conduction Block Research Articles

Ablation of hemodynamically unstable ventricular tachycardia guided by ultra-high density pacemapping and PASO correlation map, a multi-center clinical trial

Abstract Background Due to hemodynamic instability during ventricular tachycardia (VT), obtaining a complete electroanatomic map is often difficult if not impossible. Pace-mapping has progressed but still lacks precision and may not reveal sufficient details for complete ablation of the potential critical isthmus. Purpose We hypothesized that a novel strategy of combining ultra-high density (HD) pace-mapping using the PASO algorithm and Carto system to match the induced/clinical VT may aid in the reconstruction of the VT critical isthmus and effective VT ablation leading to reduce the risks of VT recurrence. Methods This was a prospective, multi-center trial conducted between January 2021 and July 2023. We enrolled 30 patients with ischemic heart disease and scar mediated recurrent VT clinically indicated for ablation. Following induction of clinical VT, we performed ultra-HD pace-mapping (5-10 mm distance between pacing points) of the identified ventricular substrate. Using PASO Module, a pacing correlation map (PCM) was constructed based on the percentage match to the induced VT. Using the PCM and the measured stimulation to electrogram intervals, the potential VT critical isthmus was reconstructed. Radiofrequency ablation was carried out to completely transect the delineated isthmus. Post ablation, repeat HD mapping and ablation was performed, until the PASO % correlation dropped from >93% to <70%, indicating at least uni-directional (orthodromic to the perceived VT circuit) conduction block through the ablation line. Follow-up data, including ICD interrogations were collected via chart review. Outcomes were compared to 60 propensity-score matched patients with an ICD who were enrolled in the Ranolazine in High-Risk Patients with Implanted Cardioverter-Defibrillators (RAID) trial who did not undergo VT ablation and served as the control group. The primary endpoint was the first occurrence of any VT at 1-year of follow-up. Results Patients enrolled had a mean age of 69 ± 9 years and 13% were women. Propensity matched patients from the RAID trial had similar baseline clinical characteristics to patients who underwent VT ablation (Table). At 1 year follow up, there was a significantly lower cumulative probability of VT/VF recurrent events in the patients who underwent PCM guided ablation as compared with the control group (17% vs 69%, respectively; log-rank p<0.001 Figure). There were no serious procedure related complications with all patients remaining free of adverse events or death at 30 days. Conclusion Our findings suggest that ablation for unstable or unmappable VT is enhanced by ultra-high density pace-mapping to reconstruct the potential VT circuit and is associated with a significantly lower rate of recurrent VT as compared with patients managed with aggressive and escalating medical therapy.Cumulative probability of ICD therapyBaseline clinical characteristics

Evaluating pro-arrhythmogenic effects of the T634S-hERG mutation: insights from a simulation study.

A mutation to serine of a conserved threonine (T634S) in the hERG K+ channel S6 pore region has been identified as a variant of uncertain significance, showing a loss-of-function effect. However, its potential consequences for ventricular excitation and arrhythmogenesis have not been reported. This study evaluated possible functional effects of the T634S-hERG mutation on ventricular excitation and arrhythmogenesis by using multi-scale computer models of the human ventricle. A Markov chain model of the rapid delayed rectifier potassium current (IKr) was reconstructed for wild-type and T634S-hERG mutant conditions and incorporated into the ten Tusscher et al. models of human ventricles at cell and tissue (1D, 2D and 3D) levels. Possible functional impacts of the T634S-hERG mutation were evaluated by its effects on action potential durations (APDs) and their rate-dependence (APDr) at the cell level; and on the QT interval of pseudo-ECGs, tissue vulnerability to unidirectional conduction block (VW), spiral wave dynamics and repolarization dispersion at the tissue level. It was found that the T634S-hERG mutation prolonged cellular APDs, steepened APDr, prolonged the QT interval, increased VW, destablized re-entry and augmented repolarization dispersion across the ventricle. Collectively, these results imply potential pro-arrhythmic effects of the T634S-hERG mutation, consistent with LQT2.

Mechanisms of pulmonary arterial hypertension-induced atrial fibrillation: insights from multi-scale models of the human atria.

This study aimed to use multi-scale atrial models to investigate pulmonary arterial hypertension (PAH)-induced atrial fibrillation mechanisms. The results of our computer simulations revealed that, at the single-cell level, PAH-induced remodelling led to a prolonged action potential (AP) (ΔAPD: 49.6 ms in the right atria (RA) versus 41.6 ms in the left atria (LA)) and an increased calcium transient (CaT) (ΔCaT: 7.5 × 10-2 µM in the RA versus 0.9 × 10-3 µM in the LA). Moreover, heterogeneous remodelling increased susceptibility to afterdepolarizations, particularly in the RA. At the tissue level, we observed a significant reduction in conduction velocity (CV) (ΔCV: -0.5 m s-1 in the RA versus -0.05 m s-1 in the LA), leading to a shortened wavelength in the RA, but not in the LA. Additionally, afterdepolarizations in the RA contributed to enhanced repolarization dispersion and facilitated unidirectional conduction block. Furthermore, the increased fibrosis in the RA amplified the likelihood of excitation wave breakdown and the occurrence of sustained re-entries. Our results indicated that the RA is characterized by increased susceptibility to afterdepolarizations, slow conduction, reduced wavelength and upregulated fibrosis. These findings shed light on the underlying factors that may promote atrial fibrillation in patients with PAH.

Determinants of electrical propagation and propagation block in Arrhythmogenic Cardiomyopathy

Gap junction and ion channel remodeling occur early in Arrhythmogenic Cardiomyopathy (ACM), but their pathogenic consequences have not been elucidated. Here, we identified the arrhythmogenic substrate, consisting of propagation slowing and conduction block, in ACM models expressing two different desmosomal gene variants. Neonatal rat ventricular myocytes were transduced to express variants in genes encoding desmosomal proteins plakoglobin or plakophilin-2. Studies were performed in engineered cells and anisotropic tissues to quantify changes in conduction velocity, formation of unidirectional propagation, cell–cell electrical coupling, and ion currents. Conduction velocity decreased by 71% and 63% in the two ACM models. SB216763, an inhibitor of glycogen synthase kinase-3 beta, restored conduction velocity to near normal levels. Compared to control, both ACM models showed greater propensity for unidirectional conduction block, which increased further at greater stimulation frequencies. Cell–cell electrical conductance measured in cell pairs was reduced by 86% and 87% in the two ACM models. Computer modeling showed close correspondence between simulated and experimentally determined changes in conduction velocity. The simulation identified that reduced cell–cell electrical coupling was the dominant factor leading to slow conduction, while the combination of reduced cell–cell electrical coupling, reduced sodium current and inward rectifier potassium current explained the development of unidirectional block. Expression of two different ACM variants markedly reduced cell–cell electrical coupling and conduction velocity, and greatly increased the likelihood of developing unidirectional block – both key features of arrhythmogenesis. This study provides the first quantitative analysis of cellular electrophysiological changes leading to the substrate of reentrant arrhythmias in early stage ACM.

Relative acceleration of orthonormal basis vectors for the geometric conduction blocks of the cardiac electric signal propagation on anisotropic curved surfaces

Geometric conduction blocks stop cardiac electric propagation due to the shape or conductivity properties of the domain. The blocks are considered to cause many abnormal cardiac electric propagations, leading to cardiac electrophysiological pathologies, such as cardiac fibrillation and arrhythmia. Locating such multidimensional conduction blocks is challenging, particularly in a complex domain with a complex shape and strong anisotropy, such as the heart. To address this problem, we propose a novel mathematical model of the geometric conduction block using the relative acceleration adopted from space-time physics. An efficient numerical scheme for the mathematical model is also proposed to predict the unidirectional conduction block effectively, even in a complex domain. The relative acceleration in the cardiac electric propagation corresponds to the sink-source relationship between the excited (after repolarization) and excitable (before depolarization) cardiac cells, representing the geometric growth rate of the volume of metric balls. The trajectory is constructed from the wavefront of diffusion-reaction equations by aligning orthonormal basis vectors along the gradient of the action potential. Relative acceleration is computed along the propagational direction from the connection 1-form of the basis vectors. The proposed mathematical model and numerical scheme are applied to demonstrate geometric conduction blocks in two-dimensional (2D) simple curved domains with strong anisotropy.

Prokaryotic voltage-gated sodium channels are more effective than endogenous Nav1.5 channels in rescuing cardiac action potential conduction: an in silico study.

Methods to augment Na+ current in cardiomyocytes hold potential for the treatment of various cardiac arrhythmias involving conduction slowing. Because the gene coding cardiac Na+ channel (Nav1.5) is too large to fit in a single adeno-associated virus (AAV) vector, new gene therapies are being developed to enhance endogenous Nav1.5 current (by overexpression of chaperon molecules or use of multiple AAV vectors) or to exogenously introduce prokaryotic voltage-gated Na+ channels (BacNav) whose gene size is significantly smaller than that of the Nav1.5. In this study, based on experimental measurements in heterologous expression systems, we developed an improved computational model of the BacNav channel, NavSheP D60A. We then compared in silico how NavSheP D60A expression vs. Nav1.5 augmentation affects the electrophysiology of cardiac tissue. We found that the incorporation of BacNav channels in both adult guinea pig and human cardiomyocyte models increased their excitability and reduced action potential duration. When compared with equivalent augmentation of Nav1.5 current in simulated settings of reduced tissue excitability, the addition of the BacNav current was superior in improving the safety of conduction under conditions of current source-load mismatch, reducing the vulnerability to unidirectional conduction block during premature pacing, preventing the instability and breakup of spiral waves, and normalizing the conduction and ECG in Brugada syndrome tissues with mutated Nav1.5. Overall, our studies show that compared with a potential enhancement of the endogenous Nav1.5 current, expression of the BacNav channels with their slower inactivation kinetics can provide greater anti-arrhythmic benefits in hearts with compromised action potential conduction.NEW & NOTEWORTHY Slow action potential conduction is a common cause of various cardiac arrhythmias; yet, current pharmacotherapies cannot augment cardiac conduction. This in silico study compared the efficacy of recently proposed antiarrhythmic gene therapy approaches that increase peak sodium current in cardiomyocytes. When compared with the augmentation of endogenous sodium current, expression of slower-inactivating bacterial sodium channels was superior in preventing conduction block and arrhythmia induction. These results further the promise of antiarrhythmic gene therapies targeting sodium channels.

PO-01-180 MECHANISMS OF PULMONARY ARTERIAL HYPERTENSION-INDUCED ATRIAL FIBRILLATION IN THE RIGHT HEART: INSIGHTS FROM MULTI-SCALE MODELS OF THE HUMAN ATRIA

Pulmonary arterial hypertension (PAH) is one of the common conditions primarily affecting the right heart and promoting atrial fibrillation (AF), but the mechanisms are poorly understood. This study aimed to use multi-scale atrial models to investigate mechanisms by which right atrial (RA) remodeling facilitates and perpetuates atrial arrhythmias. We have developed single-cell-to-tissue-level computer models to mimic control and disease conditions by incorporating the most recent experimental data on PAH-induced electrical and structural remodeling (Hiram et al., JACC, 2019). The effects of PAH-induced remodeling in both RA and left atria (LA) on action potential duration (APD), calcium transient (CaT), conduction velocity (CV), and dynamics of re-entrant excitation waves were assessed using the multi-scale computer models. Our computer simulation results suggested that at the single cell level, although PAH-induced remodeling prolonged action potential (ΔAPD: 49.6 ms in the RA vs. 41.6 ms in the LA) and increased calcium transient (ΔCaT: 7.5e-2 μM in the RA vs. 0.9e-3 μM in the LA), heterogeneous remodeling increased susceptibility to afterdepolarizations only in the RA (Fig. A). At the tissue level, dramatically reduced CV (ΔCV: -0.5 m/s in the RA vs. -0.05 m/s in the LA) abbreviated wavelength inthe RA but not inthe LA. In addition, afterdepolarizations in the RA increased repolarization dispersion and promoted unidirectional conduction block (Fig B). Finally, the upregulated fibrosis increased the chance of breakdown of excitation waves and sustained reentries in in the RA (Fig C). These results demonstrated that PAH-induced AF is mainly in RA not LA, due to a higher degree of cardiac remodeling. RA remodeling increases arrhythmias risk due to elevated susceptibility to afterdepolarizations, reduced wavelength, increased repolarization heterogeneity and upregulation of fibrosis, which together facilitate initiation and maintenance of re-entrant circuits in the right heart.

BS-454195-3 GENERATION OF A NOVEL IN-VITRO MODEL OF THE INFARCT BORDER ZONE TO STUDY HUMAN VENTRICULAR ARRHYTHMIAS

Sudden cardiac death (SCD), the majority of which is due to ventricular arrhythmias (VA). It is thought that post-myocardial infarction VA results from the presence of slowly conducting viable strands at the infarct border zone (IBZ), forming the basis for reentry We aimed to develop an in-vitro anatomical model of the IBZ to study VA. To this end, we combined advanced micropatterning and human-induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CMs) technologies hiPSC-CMs were differentiated into ventricular cells and seeded in a 2D simplified micropatterned model of IBZ. The model was created using a DMD-based maskless photolithography system (Primo). The process involves coating a culture dish with a combination of anti-fouling materials that block protein connections to the plate, a photoinitiator gel, and laser patterning of the pre-designed micro-pattern. The plate is then coated by ECM and hiPSC-CMs to generate the desired tissue geometry. Optical mapping during pacing from different sites was used to characterize the conduction properties in the model and the impact of geometry on arrhythmogenesis (Fig.1) We designed a simplified series of models of a compact scar to model VA arising at the IBZ . This model includes surviving cardiomyocyte strands within the scar. In the structural inhomogeneity group, we added a narrowing within the surviving conducting fiber (n= 18, Fig.1a), whereas the control specimens did not include any narrowing (n=12). The presence of structural inhomogeneity was found to facilitate the incidence of uni-/bi- directional conduction blocks (18/18 in the narrowing vs. 8/12 in controls, P<0.01), which also occurred at lower pacing frequencies (2.2±0.2Hz vs. 3.7±0.17Hz, P<0.001, Fig.1c-l, 2). The development of uni-directional conduction block facilitated the development of reentrant arrhythmias in the model (Fig.1e-i), with the arrhythmias developing at a greater propensity in the narrowing group (6/7 vs. 2/7, p<0.05). Repolarization analysis [action potential duration 80% (APD80) maps] showed a homogenous repolarization pattern in the model (mean APD80 standard deviation: 10±6 ms) The novel in-vitro model of the IBZ highlighted the role of structural inhomogeneities in the development of conduction blocks and reentrant arrhythmias, most probably due to source/sink mismatches, this new experimental approach may bring unique value to the study of the mechanisms and treatments of cardiac arrhythmias

PO-455617-4 GENERATION OF A NOVEL IN-VITRO MODEL OF THE INFARCT BORDER ZONE TO STUDY HUMAN VENTRICULAR ARRHYTHMIAS

Sudden cardiac death (SCD), the majority of which is due to ventricular arrhythmias (VA). It is thought that post-myocardial infarction VA results from the presence of slowly conducting viable strands at the infarct border zone (IBZ), forming the basis for reentry We aimed to develop an in-vitro anatomical model of the IBZ to study VA. To this end, we combined advanced micropatterning and human-induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CMs) technologies hiPSC-CMs were differentiated into ventricular cells and seeded in a 2D simplified micropatterned model of IBZ. The model was created using a DMD-based maskless photolithography system (Primo). The process involves coating a culture dish with a combination of anti-fouling materials that block protein connections to the plate, a photoinitiator gel, and laser patterning of the pre-designed micro-pattern. The plate is then coated by ECM and hiPSC-CMs to generate the desired tissue geometry. Optical mapping during pacing from different sites was used to characterize the conduction properties in the model and the impact of geometry on arrhythmogenesis (Fig.1) We designed a simplified series of models of a compact scar to model VA arising at the IBZ . This model includes surviving cardiomyocyte strands within the scar. In the structural inhomogeneity group, we added a narrowing within the surviving conducting fiber (n= 18, Fig.1a), whereas the control specimens did not include any narrowing (n=12). The presence of structural inhomogeneity was found to facilitate the incidence of uni-/bi- directional conduction blocks (18/18 in the narrowing vs. 8/12 in controls, P<0.01), which also occurred at lower pacing frequencies (2.2±0.2Hz vs. 3.7±0.17Hz, P<0.001, Fig.1c-l, 2). The development of uni-directional conduction block facilitated the development of reentrant arrhythmias in the model (Fig.1e-i), with the arrhythmias developing at a greater propensity in the narrowing group (6/7 vs. 2/7, p<0.05). Repolarization analysis [action potential duration 80% (APD80) maps] showed a homogenous repolarization pattern in the model (mean APD80 standard deviation: 10±6 ms) The novel in-vitro model of the IBZ highlighted the role of structural inhomogeneities in the development of conduction blocks and reentrant arrhythmias, most probably due to source/sink mismatches, this new experimental approach may bring unique value to the study of the mechanisms and treatments of cardiac arrhythmias

Mechanistic insights into spontaneous transition from cellular alternans to ventricular fibrillation.

T-wave alternans (TWA) has been used for predicting the risk of malignant cardiac arrhythmias and sudden cardiac death (SCD) in multiple clinical settings; however, possible mechanism(s) underlying the spontaneous transition from cellular alternans reflected by TWA to arrhythmias in impaired repolarization remains unclear. The healthy guinea pig ventricular myocytes under E-4031 blocking IKr (0.1μM, N=12; 0.3μM, N=10; 1μM, N=10) were evaluated using whole-cell patch-clamp. The electrophysiological properties of isolated perfused guinea pig hearts under E-4031 (0.1μM, N=5; 0.3μM, N=5; 1μM, N=5) were evaluated using dual- optical mapping. The amplitude/threshold/restitution curves of action potential duration (APD) alternans and potential mechanism(s) underlying the spontaneous transition of cellular alternans to ventricular fibrillation (VF) were examined. There were longer APD80 and increased amplitude and threshold of APD alternans in E-4031 group compared with baseline group, which was reflected by more pronounced arrhythmogenesis at the tissue level, and were associated with steep restitution curves of the APD and the conduction velocity (CV). Conduction of AP alternans augmented tissue's functional spatiotemporal heterogeneity of regional AP/Ca alternans, as well as the AP/Ca dispersion, leading to localized uni-directional conduction block that spontaneous facilitated the formation of reentrant excitation waves without the need for additional premature stimulus. Our results provide a possible mechanism for the spontaneous transition from cardiac electrical alternans in cellular action potentials and intercellular conduction without the involvement of premature excitations, and explain the increased susceptibility to ventricular arrhythmias in impaired repolarization. In this study, we implemented voltage-clamp and dual-optical mapping approaches to investigate the underlying mechanism(s) for the arrhythmogenesis of cardiac alternans in the guinea pig heart at cellular and tissue levels. Our results demonstrated a spontaneous development of reentry from cellular alternans, arising from a combined actions of restitution properties of action potential duration, conduction velocity of excitation wave and interplay between alternants of action potential and the intracellular Ca handling. We believe this study provides new insights into underlying the mechanism, by which cellular cardiac alternans spontaneously evolves into cardiac arrhythmias.

R-on-T and the initiation of reentry revisited: Integrating old and new concepts.

Initiation of reentry requires 2 factors: (1) a triggering event, most commonly focal excitations such as premature ventricular complexes (PVCs); and (2) a vulnerable substrate with regional dispersion of refractoriness and/or excitability, such as occurs during the T wave of the electrocardiogram when some areas of the ventricle have repolarized and recovered excitability but others have not. When the R wave of a PVC coincides in time with the Twave of the previous beat, this timing can lead to unidirectional block and initiation of reentry, known as the R-on-T phenomenon. Classically, the PVC triggering reentry has been viewed as arising focally from 1 region and propagating into another region whose recovery is delayed, resulting in unidirectional conduction block and reentry initiation. However, more recent evidence indicates that PVCs also can arise from the T wave itself. In the latter case, the PVC initiating reentry is not a separate event from the T wave but rather is causally generated from the repolarization gradient that manifests as the T wave. We call the former an "R-to-T" mechanism and the latter an "R-from-T" mechanism, which are initiation mechanisms distinct from each other. Both are important components of the R-on-T phenomenon and need to be taken into account when designing antiarrhythmic strategies. Strategies targeting suppression of triggers alone or vulnerable substrate alone may be appropriate in some instances but not in others. Preventing R-from-T arrhythmias requires suppressing the underlying dynamic tissue instabilities responsible for producing both triggers and substrate vulnerability simultaneously. The same principles are likely to apply to supraventricular arrhythmias.

Subthreshold delayed afterdepolarizations mediated by reduced tissue conductivity form a substrate for unidirectional block and reentry within the infarcted heart

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): British Heart Foundation, Wellcome Trust Background Delayed afterdepolarizations (DADs) due to spontaneous calcium release (SCR) events at the subcellular scale have been associated with arrhythmia formation in the border zone (BZ) of infarcted hearts. DADs may not only summate to form ectopic focal sources but may also inactivate sodium channels forming a substrate for unidirectional conduction block and reentry. The role played by infarct anatomy and altered intracellular coupling in facilitating this phenomenon is not fully understood. Purpose To use computational modelling to investigate the role of anatomical properties of the infarct BZ in creating a substrate for DAD-mediated conduction block and reentry. Methods MRI data from a porcine post-infarction heart was used to build the computational model. A phenomenological model was used to simulate SCRs in the BZ. Arrhythmia susceptibility was quantified by pacing the model followed by a pause, to see whether DADs would occur, and an extra S2 beat with different coupling intervals (CIs). Tissue conductivity in the BZ was decreased to investigate the effect of uncoupling on DAD-mediated conduction block. Results Subthreshold DADs occurring within the infarct BZ inactivated the fast sodium channels which resulted in block of S2 beats. This occurred most readily in narrow isthmuses where electrotonic load was attenuated by the non-conducting scar. DADs rendered the entire isthmus area refractory establishing a substrate for unidirectional block and reentry (see Fig. A). Reduced tissue conductivity in the BZ reduced electrotonic load on cells undergoing DADs. This led to more local tissue depolarization (Vm) as uncoupling prevented current from flowing to neighboring cells at rest (Fig. B-C). Reduced tissue conductivity also enhanced DAD-mediated block by increasing the vulnerable window for reentry initiation (700ms &amp;lt; S2 CI &amp;lt; 900ms as shown in Fig. D). Conclusion Subthreshold DADs provide a substrate for arrhythmogenesis in the infarct BZ. Tissue uncoupling enhanced the arrhythmogenic risk by increasing the time window of unidirectional block.

PO-645-01 SUBTHRESHOLD DELAYED AFTERDEPOLARIZATIONS MEDIATED BY REDUCED TISSUE COUPLING PROVIDE AN IMPORTANT SUBSTRATE FOR UNIDIRECTIONAL BLOCK AND ARRHYTHMOGENESIS IN THE INFARCT BORDER ZONE

Delayed afterdepolarizations (DADs) caused by spontaneous calcium release (SCR) events have been implicated in arrhythmia formation in the border zone (BZ) of infarcted hearts. DADs may inactivate sodium channels forming a substrate for unidirectional conduction block. The role played by infarct anatomy and altered intracellular coupling in facilitating this phenomenon is not well understood.

Pro-Arrhythmic Effects of Discontinuous Conduction at the Purkinje Fiber-Ventricle Junction Arising From Heart Failure-Induced Ionic Remodeling - Insights From Computational Modelling.

Heart failure is associated with electrical remodeling of the electrical properties and kinetics of the ion channels and transporters that are responsible for cardiac action potentials. However, it is still unclear whether heart failure-induced ionic remodeling can affect the conduction of excitation waves at the Purkinje fiber-ventricle junction contributing to pro-arrhythmic effects of heart failure, as the complexity of the heart impedes a detailed experimental analysis. The aim of this study was to employ computational models to investigate the pro-arrhythmic effects of heart failure-induced ionic remodeling on the cardiac action potentials and excitation wave conduction at the Purkinje fiber-ventricle junction. Single cell models of canine Purkinje fiber and ventricular myocytes were developed for control and heart failure. These single cell models were then incorporated into one-dimensional strand and three-dimensional wedge models to investigate the effects of heart failure-induced remodeling on propagation of action potentials in Purkinje fiber and ventricular tissue and at the Purkinje fiber-ventricle junction. This revealed that heart failure-induced ionic remodeling of Purkinje fiber and ventricular tissue reduced conduction safety and increased tissue vulnerability to the genesis of the unidirectional conduction block. This was marked at the Purkinje fiber-ventricle junction, forming a potential substrate for the genesis of conduction failure that led to re-entry. This study provides new insights into proarrhythmic consequences of heart failure-induced ionic remodeling.

Initiation and termination of reentry-like activity in rat cardiomyocytes cultured in a microelectrode array

Cardiac reentry is a lethal arrhythmia associated with cardiac diseases. Although arrhythmias are reported to be due to localized propagation abnormalities, little is known about the mechanisms underlying the initiation and termination of reentry. This is primarily because of a lack of an appropriate experimental system in which activity pattern switches between reentry and normal beating can be investigated. In this study, we aimed to develop a culture system for measuring the spatial dynamics of reentry-like activity during its onset and termination. Rat cardiomyocytes were seeded in microelectrode arrays and purified with a glucose-free culture medium to generate a culture with a heterogeneous cell density. Reentry-like activity was recorded in purified cardiomyocytes, but not in the controls. Reentry-like activity occurred by a unidirectional conduction block after shortening of the inter-beat interval. Furthermore, reentry-like activity was terminated after propagation with a conduction delay of less than 300 ms, irrespective of whether the propagation pattern changed or not. These results indicate that a simple purification process is sufficient to induce reentry-like activity. In the future, a more detailed evaluation of spatial dynamics will contribute to the development of effective treatment methods.

Does Unidirectional Block Exist after a Radiofrequency Line Creation? Insights from Ultra-High-Density Mapping (The UNIBLOCK Study).

Background: Whether unidirectional conduction block (UB) can be observed after creation of a radiofrequency (RF) line is still debated. Previous studies reported a prevalence of 9 to 33% of UB, but the assessment was performed using a point-by-point recording across the line. Ultra-high-density (UHD) system may bring some new insights on the exact prevalence of UB. Purpose: A prospective study was conducted to assess the prevalence of UB and bidirectional block (BB) using UHD system after RF line creation. Methods: Patients referred for atrial RF ablation procedure were included in this multicenter prospective study. UHD maps were performed by pacing both sides of the created line. Results: A total of 80 maps were created in 40 patients (67 ± 12 years, 70% male) by pacing (mean cycle length 600 ± 57 ms) from both sides of the cavotricuspid isthmus line. After a 47 ± 17 min waiting time after the last RF application, UHD maps (mean number of 4842 ± 5010 electrograms, acquired during 6 ± 5 min) showed that BB was unambiguously confirmed on all of them. UB was not observed in any map. After a mean follow-up of 12 ± 4 months, 6 (14%) patients experienced an arrhythmia recurrence. Conclusion: After creation of an RF line, no case of UB was observed using UHD mapping, suggesting that the presence of a conduction block along a RF line is always associated with a block in the opposite direction.

P321Subthreshold delayed afterdepolarizations form a substrate for conduction block in the infarcted heart

Abstract Funding Acknowledgements National Institute for Health Research; British Heart Foundation; and The Wellcome Trust and Engineering and Physical Sciences Research Council. Background Delayed afterdepolarizations (DADs) due to spontaneous calcium (Ca) release (SCR) events from the sarcoplasmic reticulum have been implicated with a variety of arrhythmias. Such SCR events have also been reported in cells that survive in the infarct border zone (BZ). While the potential of Ca-mediated DADs to become suprathreshold and propagate in the form of ectopic beats has been well characterized, the role of subthreshold DADs in arrhythmia formation in the infarcted heart remains to be elucidated. Purpose To use computational modelling to investigate whether subthreshold Ca-mediated DADs may form a substrate for conduction block and reentry in the BZ. Our hypothesis is that subthreshold DADs can hinder local tissue excitability in critical infarct BZ regions by inactivating the fast sodium current (INa), leading to temporary unidirectional conduction block providing a trigger for arrhythmogenesis. Methods We developed an idealized infarct model of the left ventricle. The infarct region consisted of a non-conducting scar transcended by an isthmus of cells that survived myocardial infarction (border zone). These cells were made prone to Ca-mediated DADs described by a phenomenological model of SCR events. The model was pre-paced at the apex followed by a 1500ms-pacing pause to see whether DADs would emerge. An extra beat with a longer coupling interval (CI) was then applied. The following electrophysiological changes resulting from remodeling processes in the isthmus were simulated to assess their contribution to the arrhythmogenic potential of subthreshold DADs: INa loss-of-function due to a (2.5mV and 5mV) negative-shift in the steady-state channel inactivation; 50% reduction in tissue conductivity; and increased levels of fibrosis (up to 50%). Results On average, Ca-mediated DADs reached their maximum value 1065ms after the last paced beat (Fig. A). Despite this, in the default electrophysiological setup, simulations with extra beats with 1000ms &amp;gt; CI &amp;gt; 1100ms did not result in conduction block in any of the experiments. When repeated with combined changes of reduced tissue conductivity and fibrosis, subthreshold DADs were still unable to create a substrate for block. However, when combined with a 5mV-shift in INa inactivation, block at isthmus’ mouth proximal to the stimulus site was detected for extra beats 1010 ms ≥ CI ≥ 1070ms (see Fig. B). The cause of block was due to a subthreshold DAD occurring just prior to the arrival of the extra beat. All blocked beats degenerated into reentry. Conclusions Under most physiological conditions, subthreshold DADs are unlikely to provide a substrate for unidirectional block. However, under conditions of decreased excitability, subthreshold DADs can hinder tissue excitability in the infarcted region leading to conduction block and reentry. Abstract Figure. DAD-mediated conduction block in the BZ

Validation of quantitative measure of repolarization reserve as a novel marker of drug induced proarrhythmia

Repolarization reserve, the robustness of a cell to repolarize even when one of the repolarization mechanisms is failing, has been described qualitatively in terms of ionic currents, but has not been quantified by a generic metric that is applicable to drug screening. Prolonged repolarization leading to repolarization failure is highly arrhythmogenic. It may lead to ventricular tachycardia caused by triggered activity from early afterdepolarizations (EADs), or it may promote the occurrence of unidirectional conduction block and reentry. Both types of arrhythmia may deteriorate into ventricular fibrillation (VF) and death. We define the Repolarization Reserve Current (RRC) as the minimum constant current necessary to prevent normal repolarization of a cell. After developing and testing RRC for nine computational ionic models of various species, we applied it experimentally to atrial and ventricular human induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM), and isolated guinea-pig ventricular cardiomyocytes. In simulations, repolarization was all-or-none with a precise, model-dependent critical RRC, resulting in a discrete shift in the Action Potential Duration (APD) - RRC relation, in the occurrence of EADs and repolarization failure. These data were faithfully reproduced in cellular experiments.RRC allows simple, fast, unambiguous quantification of the arrhythmogenic propensity in cardiac cells of various origins and species without the need of prior knowledge of underlying currents and is suitable for high throughput applications, and personalized medicine applications.

PITX2 upregulation increases the risk of chronic atrial fibrillation in a dose-dependent manner by modulating IKs and ICaL -insights from human atrial modelling.

BackgroundFunctional analysis has shown that the paired-like homeodomain transcription factor 2 (PITX2) overexpression associated with atrial fibrillation (AF) leads to the slow delayed rectifier K+ current (IKs) increase and the L-type Ca2+ current (ICaL) reduction observed in isolated right atrial myocytes from chronic AF (CAF) patients. Through multiscale computational models, this study aimed to investigate the functional impact of the PITX2 overexpression on atrial electrical activity.MethodsThe well-known Courtemanche-Ramirez-Nattel (CRN) model of human atrial action potentials (APs) was updated to incorporate experimental data on alterations in IKs and ICaL due to the PITX2 overexpression. These cell models for sinus rhythm (SR) and CAF were then incorporated into homogeneous multicellular one-dimensional (1D), two-dimensional (2D), and three-dimensional (3D) tissue models. The proarrhythmic effects of the PITX2 overexpression were quantified with ion current profiles, AP morphology, AP duration (APD) restitution, conduction velocity restitution (CVR), wavelength (WL), vulnerable window (VW) for unidirectional conduction block, and minimal substrate size required to induce re-entry. Dynamic behaviors of spiral waves were characterized by measuring lifespan (LS), tip patterns and dominant frequencies.ResultsThe IKs increase and the ICaL decrease arising from the PITX2 overexpression abbreviated APD and flattened APD restitution (APDR) curves in single cells. It reduced WL and increased CV at high excitation rates at the 1D tissue level. Although it had no effects on VW for initiating spiral waves, it decreased the minimal substrate size necessary to sustain re-entry. It also stabilized and accelerated spiral waves in 2D and 3D tissue models.ConclusionsElectrical remodeling (IKs and ICaL) due to the PITX2 overexpression increases susceptibility to AF due to increased tissue vulnerability, abbreviated APD, shortened WL and altered CV, which, in combination, facilitate initiation and maintenance of spiral waves.

Does unidirectional block exist after a radiofrequency line creation? Data from Ultra-High-Resolution apping (the UNIBLOCK Study)

Whether unidirectional conduction block (UB) can be observed after creation of a radiofrequency (RF) line is still a subject of debate. Previous study reported a prevalence of 9% of UB, but the assessment was performed using a point-by-point recording across the line, which may have artificially increased this UB percentage. To prospectively assess bidirectional block using ultra-high-resolution (UHR) system after RF line creation. Prospective patients admitted for an RF ablation procedure were included in this study. UHR maps were performed by pacing on both sides of the line created. A total of 18 maps were created in 8 patients (5 men, mean age 71.5 ± 10) by pacing (mean cycle length 600 ± 80 ms) from both sides of the line after a mean waiting time of 60 ± 18 min: 2 left atrial roof and 7 cavotricuspid isthmus lines; mean number of 4863 ± 3982 electrograms (381 ± 651 beats) acquired during 10 ± 9 min; mean right or left atrial volume 81 ± 82 mL. The RF lines were all unambiguously blocked bidirectionally. After a mean follow-up of 12 ± 4 months, no patient experienced any arrhythmia recurrence ( Fig. 1 ). UHR mapping confirmed that the presence of a conduction block along a RF line is always associated with a block in the opposite direction.