Human atrial electrophysiology was simulated using the Colman et al. model (Colman et al., 2013), as updated recently (Ni et al., 2017), and is hereinafter referred to as the CNZ (Colman-Ni-Zhang) model. The equations for I_Kr in the CNZ model were replaced with previously developed and validated Markov chain formulations of wild type (WT) and N588K mutant I_Kr/hERG (Whittaker et al., 2017a). The maximal conductance of I_Kr was set to g_Kr = 0.0111375·[K+]o0.59, where [K+]o is the extracellular potassium concentration (Adeniran et al., 2011), resulting in an action potential duration (APD) at 90% repolarisation (APD₉₀) of 248.8 ms at 1 Hz in the baseline human atrial cell model, which is well within the experimentally-measured range of APDs in human atrial myocytes (Bosch et al., 1999; Kim et al., 2002; Dobrev and Ravens, 2003; Katoh et al., 2005; Redpath et al., 2006; Pau et al., 2007). A family of regional human atrial cell models was incorporated into the CNZ model, accounting for distinct differences in electrophysiology of the right and left atrium (RA and LA, respectively), right and left atrial appendages (RAA and LAA, respectively), crista terminalis (CT), pectinate muscles (PM), atrio-ventricular ring (AVR), atrial septum (AS), Bachmann's bundle (BB), and pulmonary veins (PV) (Colman et al., 2013). Changes to maximal ionic conductances relative to the RA model implemented in order to create a family of regional cell models are given in Table S1. The homozygous (N588K) form of SQT1 was modeled as consisting of 100% mutant subunits, whereas the heterozygous (WT-N588K) form was assumed to consist of a 1:1 WT:mutant subunit ratio (Whittaker et al., 2017a).

In our previous study (Whittaker et al., 2017a), the actions of the class Ia anti-arrhythmic drugs disopyramide and quinidine on human ventricles were simulated in the setting of SQT1. State-dependent binding of disopyramide and quinidine to hERG channels was simulated through addition of drug-bound open and inactivated states to Markov chain formulations of I_Kr, and the guarded receptor model (Starmer et al., 1984) was used to describe use-dependent block of fast sodium current, I_Na, by both agents. Furthermore, the blocking actions of disopyramide and quinidine on slow delayed rectifier potassium current, I_Ks, L-type calcium current, I_CaL, transient outward potassium current, I_to, inward rectifier potassium current, I_K1 (quinidine only), and late sodium current, I_NaL (quinidine only), were modeled using a simple “pore block” approach. In the present study, the actions of disopyramide and quinidine on human atrial electrophysiology were represented using the same formulations and IC₅₀ (half maximal inhibitory concentration) values presented in Whittaker et al. (2017a). In addition, disopyramide and quinidine have both been reported to block the atrial-specific ultra-rapid delayed rectifier potassium current, I_Kur. For disopyramide, the IC₅₀ for block of I_Kur was taken to be 25.0 μM (Aréchiga et al., 2008), and for quinidine the IC₅₀ was taken to be 6.6 μM, as measured in human atrial myocytes (Nenov et al., 1998). As the CNZ model does not include late sodium current, quinidine block of I_NaL was not included in simulations. Full details of disopyramide and quinidine models can be found in Whittaker et al. (2017a).

Using the same approach as detailed for disopyramide and quinidine in Whittaker et al. (2017a), state-dependent models of drug block by propafenone were developed. Interactions between propafenone and I_Kr/hERG were developed based on experimental data obtained at 37°C (Paul et al., 2002; McPate et al., 2008), where estimation of parameters for drug-bound states of the Markov chain formulation of I_Kr was performed using the procedure outlined in Moreno et al. (2016). Binding and unbinding rates to activated and inactivated state channels were allowed to vary freely in order to minimize the difference between simulated and experimental dose-dependent steady state block (WT and N588K), mean fractional block of tail currents during a pulse protocol, and voltage-dependence of tail current block (Paul et al., 2002; McPate et al., 2008). The extended drug-free Markov chain model of I_Kr/hERG is shown in Figure 1, as well as the close concordance between simulated and experimental data regarding propafenone block of I_hERG. Parameters and state affinities are given in Table S2.

Propafenone is a class Ic sodium channel blocking anti-arrhythmic drug (Roden, 2014). Use-dependent block of sodium channels by propafenone was represented using the guarded receptor hypothesis (Starmer et al., 1984), as described for disopyramide and quinidine. Propafenone is predominantly an open state sodium channel blocker with little to no resting state block (Edrich et al., 2005; Burashnikov et al., 2012), and was thus assumed to bind only to activated and inactivated states. Binding and unbinding parameters were constrained based on the dose-dependent, use-dependent, and steady state block of I_Na by propafenone (Harmer et al., 2008), as shown in Figure 2A. Binding and unbinding parameters and state affinities are given in Table S3.

Whilst propafenone most potently blocks I_Kr and I_Na, it is a multi-channel blocker, and thus exerts secondary effects on other ion channels, which were modeled using a simple pore block based on IC₅₀ values from the literature. Propafenone blocks I_CaL with an IC₅₀ of 1.5–1.7 μM in mammalian cardiac myocytes (Fei et al., 1993; Hancox and Mitcheson, 1997). In human atrial myocytes, propafenone has been shown to block I_to with an IC₅₀ of 4.8 μM (Gross and Castle, 1998; Seki et al., 1999) and I_K1 with an IC₅₀ of 16.8 μM (Amorós et al., 2013). Finally, I_Kur is blocked with an IC₅₀ of 4.4 μM (Franqueza et al., 1998). The therapeutic range of propafenone has been estimated to be between 2 and 6 μM (Paul et al., 2002). Taking into account plasma protein binding, estimates of the most likely unbound concentrations have been given as 0.15–0.7 μM (Slawsky and Castle, 1994), 0.2–0.6 μM (Duan et al., 1993), and 0.33–1 μM (Seki et al., 1999). In this study, the effects of three equally-spaced concentrations which fell within this range were studied−0.2, 0.5, and 0.8 μM propafenone. It should be noted that these concentrations are not intended to be compared directly with those chosen for disopyramide and quinidine (1, 2, and 5 μM), which were taken from Whittaker et al. (2017a) and are not equally-spaced. A comparison of IC₅₀ values used for disopyramide, quinidine, and propafenone is given in Table S4.

In order to assess whether the combined actions of propafenone in the model induced reasonable rate- and concentration- dependent effects on the AP under control conditions, alterations in the maximum upstroke velocity (V°max) upon application of propafenone were compared with experimental data taken from canine atrial cells (Burashnikov et al., 2012). Figure 2B shows changes in V°max upon application of 0.3 and 1.5 μM propafenone at cycle lengths of 500 and 300 ms, where close concordance can be seen between simulation and experiment (Figure 2C). Although experimental data were taken from canine atrial myocytes, the human atrial simulation data nonetheless recapitulated the considerable reduction in V°max at both concentrations and cycle lengths, which is primarily due to the sodium channel blocking actions of propafenone.

Propagation of excitation waves in tissue was described using the monodomain equation, (1)∂V∂t=∇(D∇V)-IionCm,

where V is the transmembrane voltage, D is the global conductivity tensor, I_ion is the total ionic current, and C_m is the membrane capacitance. Equation (1) was solved using a finite difference PDE solver based on the explicit forward Euler method and Strang splitting scheme. Effects of the N588K-hERG mutation on ERP, WL, conduction velocity (CV), and spiral wave dynamics were determined using 1D and 2D models of RA tissue, as described previously (Whittaker et al., 2017b, 2018). Spiral waves were initiated in 2D models using an S1-S2 cross-shock protocol, and spiral wave core trajectories over a 5.0 s simulation period were traced by locating phase singularities (Bray and Wikswo, 2002). The behavior of re-entrant excitations in an anatomically-realistic setting was determined using a 3D human atrial structured grid geometry based on the Visible Female dataset (Seemann et al., 2006; Colman et al., 2013; Whittaker et al., 2017b) with rule-based fiber orientations (Krueger et al., 2011). The transverse value of the diffusion coefficient, D_⊥, was set to 0.1 mm² ms⁻¹, where an anisotropy ratio (D_||:D_⊥) of 3:1 in directions longitudinal and transverse to fibers was applied in the atrial working myocardium, and a ratio of 9:1 along the fast conducting bundles of the BB, CT, and PM (Colman et al., 2013; Whittaker et al., 2017b), which gave global and specific regional activation times in close agreement with experimental measurements (Lemery et al., 2007). The activation (ACT) time was defined as the time required for the membrane potential to reach −20 mV at each point in the geometry (Ni et al., 2017). Activation-recovery interval (ARI) was defined as the time interval between membrane potential depolarisation to −20 mV and repolarisation to −70 mV (adjusted to −65 mV in the PV region due to higher resting membrane potential) (Ni et al., 2017).

Re-entry was initiated using the phase distribution method (Biktashev and Holden, 1998; Whittaker et al., 2017a), by initiating either a clockwise or anti-clockwise scroll wave (as seen from a RA posterior wall view). Where sustained re-entrant activity was initiated, a power spectrum was obtained through fast Fourier transform analysis of pseudo ECG (pECG) time series (recorded from within the RA cavity). The dominant frequency (DF) was computed in Matlab from the largest peak in the power spectrum density. For simulating the effects of disopyramide, quinidine, and propafenone on re-entrant excitation, state variables for each node within the 3D anatomical model were saved after 2.5 s of a 10.0 s episode of sustained re-entrant activity in drug-free conditions. These were then used as initial conditions for new 3D simulations of duration 7.5 s (giving 10.0 s activity overall) in which varying concentrations of anti-arrhythmic drugs (1, 2, or 5 μM disopyramide and quinidine; 0.2, 0.5, or 0.8 μM propafenone) were applied immediately. This gave 18 simulations in total; 3 doses for each of the 3 anti-arrhythmic drugs, with 2 scroll wave initial conditions (both clockwise and anti-clockwise).

Both SQT1 mutant conditions shortened the human atrial APD at a pacing rate of 1 Hz (Figure 3Ai), from 248.8 ms in the WT condition to 195.4 and 163.3 ms in heterozygous (WT-N588K) and homozygous (N588K) mutation conditions, respectively. This was due to loss of inactivation associated with the N588K-hERG mutation, which increased I_Kr early during the AP and served to accelerate the repolarisation process (Figure 3Aii). Action potential abbreviation reduced considerably the duration of the plateau phase, causing I_K1 to contribute to terminal repolarisation earlier during the AP (Figure 3Aiii), and lesser activation and reduced contribution of I_Ks (Figure 3Aiv). The SQT1 mutation also decreased net I_CaL due to the abbreviated plateau phase (Figure 3Av), shortening the APD₉₀ under heterozygous and homozygous conditions (Figure 3Avi). In tissue, accelerated repolarisation under SQT1 conditions shortened the ERP and consequently the excitation WL. At the organ scale (in the 3D anatomical human atria model), both heterozygous and homozygous forms of the N588K-hERG mutation decreased global ARI (Figure 3B), whilst preserving the global dispersion of ARI. Furthermore, whereas ΔARI was decreased between CT/PM regions, it was increased between PV/LA and RA/LA regions under SQT1 conditions (Figure 3C). A summary of the effects of the N588K-hERG mutation on multi-scale human atrial AP biomarkers is given in Table 1.

A summary of 3D scroll wave simulations in the anatomical human atria model under WT and SQT1 mutation conditions (in a clockwise configuration from a RA posterior wall aspect) is given in Figure 4. In the WT condition, scroll waves followed 2 transient, fragmented circuits around the RA, before self-terminating at ~0.7 s (Video S1), which precluded accurate measurement of the DF. In the WT-N588K condition, re-entrant wave activity sustained for the entire 10.0 s with a DF of 4.99 Hz, and was driven mostly by a macro re-entrant circuit around the right AVR, whilst also showing CT/isthmus driven activity in the RA (Video S2). Under homozygous N588K mutation conditions, scroll wave activity was also driven predominantly by a re-entrant circuit around the AVR, with occasional existence of multiple waves on the RA free wall (Video S3), and a DF of 5.59 Hz. A summary of simulations using anti-clockwise scroll wave initial conditions is shown in Figure S1, and an average of the DF from both simulations (where applicable) measured from the first 5.0 s of activity is given in Table 1.

In order to assess the actions of class I anti-arrhythmic drugs on the human atria at the cellular level, the effects of applying various concentrations of disopyramide, quinidine, and propafenone were investigated on the single cell AP under heterozygous WT-N588K (hereinafter referred to simply as SQT1) conditions. Figure 5 shows AP profiles under drug-free SQT1 conditions and upon application of different concentrations of each drug at a pacing frequency of 1 Hz, with corresponding fractional block of I_Kr and I_Na. Application of all concentrations of disopyramide and propafenone produced only modest prolongation of the APD, failing to restore it to that of the WT condition. Quinidine, on the other hand, was more effective at prolonging the APD due to extensive I_Kr block, with the highest concentration (5 μM) prolonging the APD beyond that of the WT level. In contrast, all three agents prolonged the ERP effectively, restoring it to (or exceeding) that of the WT level at the highest respective concentrations tested, due to additional ERP-prolonging effects of I_Na block.

Conduction slowing due to sodium channel block by the three drugs caused an increase in activation time in the 3D anatomical human atria model (Figure 5D), which was most prominent for quinidine and least prominent for propafenone at the concentrations tested. A summary of the effects of pharmacological modulation across multiple scales is given in Table 2. In addition, in the 1D tissue model, the rate-dependent effects of disopyramide, quinidine, and propafenone on the ERP, CV, and WL are shown in Figure S2. Quinidine was shown to produce the most potent effects on CV and ERP at all pacing rates. Propafenone exerted the weakest effects on ERP at fast pacing rates, but had the greatest propensity to promote beat-to-beat alternans.

In 2D simulations, under drug-free WT conditions the initiated spiral wave failed to re-enter the tissue (not shown). Under drug-free SQT1 conditions, the initiated spiral wave sustained for the 5.0 s duration of the simulation, eventually settling into a stationary, epicycloidal trajectory (Figure 6Ai). Application of 5 μM disopyramide was sufficient to terminate re-entry, as the re-entry wavelength was increased to such an extent that the spiral wave meandered out of the tissue (Figure 6Aii). On the other hand, both 1 and 2 μM disopyramide did not terminate re-entrant activity, but did increase the area over which spiral waves meandered (from 0.35 mm² ms⁻¹ in the WT condition to 0.40 and 0.47 mm² ms⁻¹, respectively '(Figure 6B). Similarly, 1 μM quinidine increased the area of meander to 0.44 mm² ms⁻¹ without terminating re-entry, whereas 2 and 5 μM quinidine terminated re-entry without inducing wave break (Figure 6C). Application of 0.2 and 0.5 μM propafenone resulted in termination of re-entry by decreasing both the stationarity and stability of spiral waves (Figure 6D), whereas 0.8 μM propafenone did not terminate re-entry but did destabilize the re-entrant circuit and substantially increase the area of meander (to 0.75 mm² ms⁻¹).

In 3D simulations using clockwise scroll wave initial conditions, quinidine terminated re-entry at all concentrations, and disopyramide terminated re-entry at concentrations of 1 and 5 μM (arrhythmia termination by 5 μM disopyramide and 2 μM quinidine are shown in Videos S4, S5, respectively). At a concentration of 2 μM disopyramide, the DF was reduced from 4.79 to 3.99 Hz (measured from the final 5.0 s of activity). Propafenone, on the other hand, was comparatively ineffective at reducing the DF at the concentrations tested, but terminated re-entry at a concentration of 0.5 μM (Video S6). Representative examples of arrhythmia termination by disopyramide, propafenone, and quinidine are shown in Figure 7. Disopyramide and quinidine produced a dose-dependent decrease in the DF which was greater than for propafenone (Figure 7C). In an anti-clockwise scroll wave configuration, the efficacy of anti-arrhythmic drugs was less favorable, with pharmacological conversion of re-entrant waves occurring only for 0.5 μM propafenone and 5 μM quinidine. A quantitative summary of all 2D and 3D re-entry simulations is given in Table S5.

The major findings presented in this study are as follows. (1) Heterozygous and homozygous forms of the SQT1-linked N588K-hERG mutation shortened significantly the APD of human atrial cells, whilst preserving global dispersion of ARI, and increasing ΔARI between certain regions of the human atria. (2) The dominant frequency and lifespan of re-entry in 3D arrhythmia simulations was increased by heterozygous and homozygous forms of the N588K mutation. (3) Disopyramide, quinidine, and propafenone all produced ERP prolongation in the setting of SQT1, the extent of which was greatest for quinidine. (4) In 2D simulations, disopyramide and quinidine terminated re-entry at high concentrations due to increased re-entry wavelength, whereas propafenone terminated re-entry in a non-dose-dependent manner, by inducing secondary waves. (5) In 3D simulations, the DF of re-entry was reduced in a dose-dependent manner for clinically-relevant doses of disopyramide and quinidine, and propafenone to a lesser extent. (6) All three anti-arrhythmic agents demonstrated some efficacy in pharmacological rhythm control, most frequently terminating re-entry in the order quinidine > propafenone = disopyramide. A summary of findings regarding pro-arrhythmic mechanisms of the N588K-hERG mutation in human atria and anti-arrhythmic actions of selected class I drugs is given in Figure 8.

The fact that the N588K-hERG mutation is associated with both a short QT interval and reports of AF (Hong et al., 2005) suggests that pathophysiological mechanisms leading to QT interval shortening in SQT1 also affect the atria and may promote AF. In an experimental model of SQT1, application of 20 μM of the I_Kr agonist PD-118057 shortened the APD in both CT and PM cell types of canine atrial tissue (Nof et al., 2010), as well as increasing APD dispersion between them. In the present study, at the cellular level heterozygous and homozygous forms of the N588K-hERG mutation produced a shortening of the human right atrial APD of 53.4 and 85.5 ms, respectively–values which are consistent with a previous simulation study (Loewe et al., 2014b). At the tissue level, global ΔARI was unaffected by the WT-N588K mutation condition, and was slightly decreased by the homozygous form. The model predicted an increase in ΔARI between PV/LA and RA/LA regions for both heterozygous and homozygous forms of the N588K mutation (Figure 3C), but not between CT/PM regions. This may be due to the fact that the effects of the hERG activator PD-118057 and of the N588K mutation on I_Kr/hERG are different: the former increases I_Kr/hERG channel open probability without altering gating kinetics (Zhou et al., 2005), whereas the latter alters kinetics through profound attenuation of inactivation (Cordeiro et al., 2005; McPate et al., 2005).

Interestingly, there is clinical evidence of a LA-RA gradient in the DF in paroxysmal AF (Lazar et al., 2004), which may be underlain by APD differences between the RA and LA in pathological conditions which promote AF (Voigt et al., 2010). Furthermore, regional differences in APD between PV/LA regions have been identified previously as high frequency excitation or microreentrant sources underlying AF (Mandapati et al., 2000; Arora et al., 2003; Kumagai et al., 2004)–this suggests that localized increases in the spatial dispersion of repolarisation could be a mechanism by which atrial arrhythmogenesis is promoted in SQT1, as suggested in the previous experimental study (Nof et al., 2010).

In a previous simulation study (Loewe et al., 2014b), the N588K-hERG mutation in a 1:1 mutant to WT ratio was shown to increase inducibility and lifespan of rotors in idealized 2D sheets of human atrial tissue, but did not permit sustained re-entry. By contrast, in the present study, the heterozygous WT-N588K (SQT1) condition permitted the sustenance of spiral waves in 2D simulations. This may be due to several differences in the modeling, one of which is that the CRN model used in that study (Loewe et al., 2014b) produces an ERP restitution curve which is around 50 ms higher than available experimental data (Wilhelms et al., 2013), making re-entry more difficult to sustain, whereas the CNZ model ERP restitution curve lies mostly within the experimental range (Figure S3). In this study, the presence of realistic structure and heterogeneous electrophysiology, in addition to the cellular electrophysiological substrate induced by the N588K-hERG mutation, also favored the sustenance of re-entrant scroll waves in the 3D anatomical human atria model (using both clockwise and anti-clockwise scroll wave initial conditions), and could both prolong or shorten the lifespan of re-entrant excitations under pharmacological modulation conditions when compared directly to observations in 2D homogeneous sheets. This highlights the value of including both approaches in probing the arrhythmia substrate associated with genetic mutations in human atria, as behavior in heterogeneous 3D anatomical models cannot necessarily be predicted from homogeneous 2D simulations, and vice versa. It should be noted nonetheless that electrical activity observed in 3D re-entry simulations in this study remained relatively organized and generally not driven by multiple wavelets. This lack of complex, chaotic behavior which is observed in persistent AF is consistent with reports of paroxysmal AF in some patients with the N588K-hERG mutation (Hong et al., 2005) – addition of electrical and intracellular gap junction remodeling or fibrosis in patients may be required to promote degeneration of paroxysmal AF into persistent AF.

Despite the prevalence of AF and decades of research, anti-arrhythmic therapies for AF continue to have limited efficacy and safety (Workman et al., 2011). In this study, the class I anti-arrhythmic drugs disopyramide, quinidine, and propafenone were shown to be only moderately effective in the management of atrial arrhythmias mediated by the SQT1-linked N588K-hERG mutation, although it should be noted that the 7.5 s duration of 3D drug simulations is much shorter than clinical time scales. Pharmacological rhythm control occurred under application of both disopyramide and quinidine, but in an unpredictable, non-dose-dependent way, as observed in our previous in silico study (Whittaker et al., 2017a). One mechanism of re-entry termination involving reduction of the excitable gap under application of 5 μM quinidine is shown in Figure S4.

Both disopyramide and quinidine reduced consistently the DF of re-entry in a dose-dependent manner, although as in (Whittaker et al., 2017a) quinidine was more effective than disopyramide at rate control in the setting of SQT1, likely due to its more potent block of I_Kr (McPate et al., 2008) and I_Na (Koumi et al., 1992). Propafenone was comparatively ineffective at controlling the rate of atrial arrhythmias under the WT-N588K mutation condition, although did demonstrate some efficacy in re-entry termination. Figure S5 shows a mechanism of pharmacological rhythm control by 0.5 μM propafenone, in which secondary waves induced by propafenone extinguished all re-entrant pathways. This is consistent with the use of propafenone to manage paroxysmal AF in SQT1 patients (Hong et al., 2005; Bjerregaard et al., 2006), which maintained 3 patients free of arrhythmia recurrence during follow up. However, as propafenone has been reported to have no prolonging effect on the QT interval in the setting of SQT1 (Hong et al., 2005)— an effect which was demonstrated with variable success using two leading human ventricle models (ten Tusscher and Panfilov, 2006; O'Hara et al., 2011) in Figure S6—it is likely that quinidine and disopyramide will remain more suitable pharmacotherapies.

In this study, cellular and tissue level simulations (in a 1D model) alone did not serve as good predictors of the overall effects of anti-arrhythmic drugs in the electrically-heterogeneous, 3D anatomical whole atria model, consistent with previous modeling studies (Varela et al., 2016; Whittaker et al., 2017b). For example, disopyramide was shown not to strongly affect the excitation WL in RA tissue (Table 2; Figure S2), and yet it decreased reliably the DF of re-entrant excitations under heterozygous N588K mutation conditions in 3D simulations. This is likely due to the fact that scroll waves were largely anatomically driven by a typical macro re-entrant atrial flutter/tachycardia circuit (Lee et al., 2012) which was larger than the functional WL; effects of disopyramide on CV were thus more important. In contrast, propafenone produced only modest changes to the CV and was thus less effective at rate control in the model, but did terminate re-entry under conditions in which the re-entrant circuit was abolished by interaction with secondary waves (Figure S5).

Further support for quinidine as a useful pharmacotherapy for AF in the setting of SQT1 comes from the experimental use of the I_Kr agonist PD-118057 model (Nof et al., 2010). That study suggested that the combined blocking actions on I_Kr and I_Na are what made quinidine effective at preventing sustained AF in canine right atrial tissue preparations, as neither I_Kr nor I_Na block alone (by E-4031 or lidocaine, respectively) was effective at terminating AF. Similarly, in our previous simulation study of ventricular pharmacotherapy for SQT1 we showed that combined block of I_Na and I_Kr by disopyramide and quinidine prolonged the ERP to a greater extent than block of I_Na or I_Kr alone (Whittaker et al., 2017a). In this study, we sought to determine whether the greater ERP prolongation associated with the synergistic combination of I_Na and I_Kr block by class I drugs would also translate to greater efficacy in terminating re-entry in a 2D sheet of SQT1 mutant human atrial tissue. Figure S7 shows that the actions of 5 μM of a hypothetical drug with disopyramide kinetics for I_Na and I_Kr block were able to terminate re-entry, whereas I_Na or I_Kr block alone did not, as combined block produced the greatest prolongation of the ERP. The spiral wave trajectories highlight the fact that I_Na block in particular destabilized the re-entrant circuit, which, when combined with I_Kr block, caused the spiral wave to meander out of the tissue boundaries (Video S7). The findings from in silico and in vitro experiments thus suggest that the combination of prolonged APD and refractoriness arising from K⁺ and Na⁺ channel inhibition is likely to be valuable in the setting of SQT1. Though selective blockers of I_Ks and I_K1 are not yet clinically-available, this combination may also warrant investigation as a pharmacotherapeutic strategy for other forms of K⁺-linked SQTS (SQT2 and SQT3).

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.