scholarly journals GPER mediated  estrogenic amelioration of sodium channel dysfunction in stressed human induced  pluripotent stem cell-derived cardiomyocytes            

2021 ◽  
Author(s):  
Xide Hu ◽  
Lu Fu ◽  
Mingming Zhao ◽  
Hongyuan Zhang ◽  
Zheng Gong ◽  
...  
Keyword(s):  
Stem Cell ◽  
Sodium Channel ◽  
Sodium Channels ◽  
Small Interfering Rna ◽  
Pluripotent Stem Cell ◽  
Sodium Current ◽  
Induced Pluripotent Stem Cell ◽  
Sodium Currents ◽  
Induced Pluripotent ◽  
Interfering Rna

Stress-induced excessive activation of the adrenergic system or changes in estrogen levels promote the occurrence of arrhythmias. Sodium channel, a responder to β-adrenergic stimulation, is involved in stress-induced cardiac electrophysiological abnormalities. However, it has not been established whether estrogen regulates sodium channels during acute stress. Our study aimed to explore whether voltage-gated sodium channels play roles in the rapid regulation of various concentrations of estrogen in stressed human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), and reveal the possible mechanism of estrogen signaling pathway modulating stress. An isoproterenol-induced stress model of hiPSC-CMs was pre-incubated with β-Estradiol at different concentrations (0.01 nmol/L, 1 nmol/L, and 100 nmol/L). Action potential (AP) and sodium currents were detected by patch clamp. The G protein-coupled estrogen receptor (GPER)-specific effect was determined with agonists G1, antagonists G15 and small interfering RNA. β-Estradiol at concentrations of 0.01 nmol/L, 1 nmol/L, and 100 nmol/L increased the peak sodium current and prolonged AP duration (APD) at 1 nmol/L. Stress increased peak sodium current, late sodium current, and shortened APD. The effects of stress on sodium currents and APD were eliminated by β-Estradiol. Activation of GPER by G1 exhibited similar effects as β-Estradiol, while inhibition of GPER with G15 and small interfering RNA ameliorated estrogenic actions. Estrogen, antagonized the stress-related abnormal electrical activity, and through GPER alleviated sodium channel dysfunctions in stress state in hiPSC-CMs. These results provide a novel mechanism through which estrogenic rapid signaling against stress by regulating ion channels.

scholarly journals GS-967 and Eleclazine Block Sodium Channels in Human Induced Pluripotent Stem Cell-derived Cardiomyocytes

2020 ◽  
Author(s):  
Franck Potet ◽  
Defne E. Egecioglu ◽  
Paul W. Burridge ◽  
Alfred L. George
Keyword(s):  
Stem Cell ◽  
Sodium Channel ◽  
Sodium Channels ◽  
Pluripotent Stem Cell ◽  
Sodium Current ◽  
Induced Pluripotent Stem Cell ◽  
Slow Inactivation ◽  
Molecular Pharmacology ◽  
Recovery From Inactivation ◽  
Induced Pluripotent

ABSTRACTGS-967 and eleclazine (GS-6615) are novel sodium channel inhibitors exhibiting antiarrhythmic effects in various in vitro and in vivo models. The antiarrhythmic mechanism has been attributed to preferential suppression of late sodium current (INaL). Here, we took advantage of a throughput automated electrophysiology platform (SyncroPatch 768PE) to investigate the molecular pharmacology of GS-967 and eleclazine on peak sodium current (INaP) recorded from human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes. We compared GS-967 and eleclazine to the antiarrhythmic drug lidocaine, the prototype INaL inhibitor ranolazine, and the slow inactivation enhancing drug lacosamide. In human induced pluripotent stem cell-derived cardiomyocytes, GS-967 and eleclazine caused a reduction of INaP in a frequency-dependent manner consistent with use-dependent block (UDB). GS-967 and eleclazine had similar efficacy but evoked more potent UDB of INaP (IC50=0.07 and 0.6 μM, respectively) than ranolazine (7.8 μM), lidocaine (133.5 μM) and lacosamide (158.5 μM). In addition, GS-967 and eleclazine exerted more potent effects on slow inactivation and recovery from inactivation compared to the other sodium channel blocking drugs we tested. The greater UDB potency of GS-967 and eleclazine was attributed to the significantly higher association rates (KON) and moderate unbinding rate (KOFF) of these two compounds with sodium channels. We propose that substantial UDB contributes to the observed antiarrhythmic efficacy of GS-967 and eleclazine.SIGNIFICANCE STATEMENTWe investigated the molecular pharmacology of GS-967 and eleclazine on sodium channels in human induced pluripotent stem cell derived cardiomyocytes using a high throughput automated electrophysiology platform. Sodium channel inhibition by GS-967 and eleclazine has unique features including accelerating the onset of slow inactivation and impairing recovery from inactivation. These effects combined with rapid binding and moderate unbinding kinetics explain potent use-dependent block, which we propose contributes to their observed antiarrhythmic efficacy.

scholarly journals Hyperthermia Influences the Effects of Sodium Channel Blocking Drugs in Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes

PLoS ONE ◽  
2016 ◽  
Vol 11 (11) ◽  
pp. e0166143 ◽  
Author(s):  
Ibrahim El-Battrawy ◽  
Siegfried Lang ◽  
Zhihan Zhao ◽  
Ibrahim Akin ◽  
Gökhan Yücel ◽  
...  
Keyword(s):  
Stem Cell ◽  
Sodium Channel ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Channel Blocking ◽  
Induced Pluripotent

A cellular model of Brugada syndrome with SCN10A variants using human-induced pluripotent stem cell-derived cardiomyocytes

EP Europace ◽  
2019 ◽  
Vol 21 (9) ◽  
pp. 1410-1421 ◽  
Author(s):  
Ibrahim El-Battrawy ◽  
Sebastian Albers ◽  
Lukas Cyganek ◽  
Zhihan Zhao ◽  
Huan Lan ◽  
...  
Keyword(s):  
Stem Cell ◽  
Sodium Channel ◽  
Brugada Syndrome ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Dermal Fibroblasts ◽  
Cellular Model ◽  
Channel Current ◽  
Healthy Control ◽  
Induced Pluripotent

Abstract Aims Brugada syndrome (BrS) is associated with a pronounced risk to develop sudden cardiac death (SCD). Up to 21% of patients are related to mutations in SCN5A. Studies identified SCN10A as a contributor of BrS. However, the investigation of the human cellular phenotype of BrS in the presence of SCN10A mutations remains lacking. The objective of this study was to establish a cellular model of BrS in presence of SCN10A mutations using human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Methods and results Dermal fibroblasts obtained from a BrS patient suffering from SCD harbouring the SCN10A double variants (c.3803G>A and c.3749G>A) and three independent healthy control subjects were reprogrammed to hiPSCs. Human-induced pluripotent stem cells were differentiated into cardiomyocytes (hiPSC-CMs).The hiPSC-CMs from the BrS patient showed a significantly reduced peak sodium channel current (INa) and a significantly reduced ATX II (sea anemone toxin, an enhancer of late INa) sensitive as well as A-887826 (a blocker of SCN10A channel) sensitive late sodium channel current (INa) when compared with the healthy control hiPSC-CMs, indicating loss-of-function of sodium channels. Consistent with reduced INa the action potential amplitude and upstroke velocity (Vmax) were significantly reduced, which may contribute to arrhythmogenesis of BrS. Moreover, Ajmaline effects on action potentials were stronger in BrS-hiPSC-CMs than in healthy control cells. This is in agreement with the higher susceptibility of patients to sodium channel blocking drugs in unmasking BrS. Conclusion Patient-specific hiPSC-CMs are able to recapitulate single-cell phenotype features of BrS with SCN10A mutations and may provide novel opportunities to further elucidate the cellular disease mechanism.

Abstract 16472: Deciphering a Mechanistic Link Between Enhanced Late Sodium Current and Triggered Atrial Fibrillation Using Patient-specific and Gene-corrected Induced Pluripotent Stem Cell-derived Atrial Cardiomyocytes

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Liang HONG ◽  
Olivia T Ly ◽  
Hanna Chen ◽  
Arvind Sridhar ◽  
Meihong Zhang ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Atrial Fibrillation ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Sodium Current ◽  
Induced Pluripotent Stem Cell ◽  
Patient Specific ◽  
Gain Of Function ◽  
Late Sodium Current ◽  
Induced Pluripotent

Introduction: Gain-of-function mutations in SCN5A, which encodes the cardiac sodium channel, have been linked with familial atrial fibrillation (AF). However, the mechanistic link between the late sodium current (I Na,L ) and triggered arrhythmia remains unclear. Hypothesis: To characterize the electrophysiological (EP) phenotype of gain-of-function AF-linked SCN5A mutations, elucidate the underlying cellular mechanisms using patient-specific and gene-corrected (GC) induced pluripotent stem cell-derived atrial cardiomyocytes (iPSC-aCMs). Methods: We generated iPSC-aCMs from two families carrying SCN5A mutations (E428K and N470K) and control subjects. Whole-cell patch clamp and multi-electrode arrays were recorded to assess the EP phenotypes of the atrial iPSC-CMs. We corrected the E428K iPSC-aCMs using CRISPR/Cas9 gene editing approach (isogenic control). Results: The SCN5A mutation lines displayed abnormal EP properties including increased beating frequency and irregularity with triggered beats characteristic of AF ( Fig. 1 ). E428K iPSC-aCMs displayed spontaneous arrhythmogenic activity with beat-to-beat irregularity ( Fig. 1 A-D ) with the prolonged APD ( Fig. 1 E-H ) associated with enhanced I Na,L ( Fig. 1 I-L ). In contrast, expression of SCN5A -E428K in heterologous expression system failed to show enhanced I Na,L . The gene-corrected E428K iPSC-aCMs normalized the aberrant EP phenotype. Gene expression profiling revealed differential expression of calcium and potassium channel homeostasis and nitric oxide mediated signal transduction which could result in EP remodeling of atrial CMs. Conclusions: Patient-specific and gene-corrected iPSC-aCMs exhibited striking ex-vivo EP phenotype of an AF-causing SCN5A gain-of-function mutation that produced minimal changes in-vitro . We established a mechanistic link between enhanced I Na,L , ion channel remodeling and nitric oxide signaling pathways, and triggered AF.

Abstract 384: Evaluating Late Sodium Current in Human Induced Pluripotent Stem Cell Derived Cardiomyocytes

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Derek Schocken ◽  
Jayna Stohlman ◽  
Xiaoyu Zhang ◽  
Yama Abassi ◽  
Lars Johannesen ◽  
...  
Keyword(s):  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Sodium Current ◽  
Field Potential ◽  
Induced Pluripotent Stem Cell ◽  
Dependent Manner ◽  
Late Sodium Current ◽  
Beating Rate ◽  
Induced Pluripotent ◽  
Dose Dependent

Background: Inhibiting late sodium current (I NaL ) reduced drug-induced QTc prolongation in a recent clinical trial. Induced pluripotent stem cell derived cardiomyocytes (iPSC-CMs) have emerged as a valuable tool in preclinical assessment of multichannel blocking drugs’ potential to prolong QT and induce arrhythmias. However, sodium channels in commercially available iPSC-CMs are known to be under expressed, necessitating investigation into the presence and effects of I NaL in this electrophysiological model. Methods and Results: A platform combining simultaneous measurements of field potential and contraction (xCELLigence RTCA CardioECR, ACEA Biosciences) was used to assess the acute effects of three I NaL enhancing drugs, ATX-II, ibutilide, and alfuzosin, given alone or in combination with an I NaL blocker, lidocaine in iPSC-CMs (iCell Cardiomyocytes 2 , Cellular Dynamics). Additionally, dofetilide, diltiazem, and lidocaine alone were included as positive controls for hERG, L-type calcium, and sodium channel block. ATX-II, a potent and specific I NaL enhancer, caused significant dose dependent rate-corrected field potential duration (FPDc) prolongation, which was then subsequently reduced in a dose dependent manner by the addition of lidocaine. At 100 nM ATX-II prolonged the FPDc by 1153.8 ± 135.8 ms from 360.5 ± 16.4 ms at the baseline, which was then reduced to 537 ± 37.4 ms with the addition of 30 μM lidocaine. Ibutilide (0.1-1 μM), a class III antiarrhythmic, caused beating rate decreases and early after depolarizations (EADs) that were not affected by lidocaine addition. Alfuzosin, which increases both peak and late sodium currents, caused dose-dependent reduction of beating rate, FPDc prolongation, and EADs at 5 μM and 10 μM. Alfuzosin-induced EADs were mitigated by addition of lidocaine (5-15 μM). Conclusions: Late sodium current enhancers prolonged repolarization and induced arrhythmias in human iPSC-CMs. These effects were reversed by addition of lidocaine, a specific late sodium current blocker. These results are consistent with the late sodium current being present in iPSC-CMs in the presence of a late sodium current enhancer, which may have implications for drug safety testing.

scholarly journals GS-967 and Eleclazine Block Sodium Channels in Human Induced Pluripotent Stem Cell–Derived Cardiomyocytes

2020 ◽  
Vol 98 (5) ◽  
pp. 540-547
Author(s):  
Franck Potet ◽  
Defne E. Egecioglu ◽  
Paul W. Burridge ◽  
Alfred L. George
Keyword(s):  
Stem Cell ◽  
Sodium Channels ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Induced Pluripotent
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