Patient-Specific Induced Pluripotent Stem-Cell Models for Long-QT Syndrome

Background - Prior epidemiological studies demonstrated that the p.D85N-KCNE1 common variant reduces repolarization reserve and predisposes to drug-induced QT prolongation/torsades de pointes. We sought to develop a cellular model for drug-induced long QT syndrome (DI-LQTS) using a patient-specific induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM). Methods - p.D85N-KCNE1 iPSCs were generated from a 23-year-old female with an exaggerated QTc response to metoclopramide (ΔQTc of 160 ms). CRISPR/Cas9 technology was used to generate "gene-corrected" isogenic iPSCs. Field potential duration (FPD) and action potential duration (APD) were measured from iPSC-CMs. Results - At baseline, p.D85N-KCNE1 iPSC-CMs displayed significantly longer FPD (281 ± 15 ms, n=13 vs. 223 ± 8.6 ms, n=14, p<0.01) and APD 90 (579 ± 22 ms, n=24 vs. 465 ± 33 ms, n=26, p<0.01) than isogenic-control iPSC-CMs. Dofetilide at a concentration of 2nM increased significantly FPD (379 ± 20 ms, n=13, p<0.01) and APD 90 (666 ± 11 ms, n=46, p<0.01) in p.D85N-KCNE1 iPSC-CMs, but not in isogenic-control. The effect of dofetilide on APD 90 (616 ± 54 ms, n=7 vs. 526 ± 54 ms, n=10, p<0.05) was confirmed by Patch-clamp. Interestingly, treatment of p.D85N-KCNE1 iPSC-CMs with estrogen at a concentration of 1nM exaggerated further dofetilide-induced APD 90 prolongation (696 ± 9 ms, n=81, p<0.01) and caused more early afterdepolarizations (EADs) (11.7%) compared to isogenic control (APD 90 : 618 ± 8 ms, n=115 and EADs: 2.6%, p<0.05) Conclusions - This iPSC-CM study provides further evidence that the p.D85N-KCNE1 common variant in combination with environmental factors such as QT prolonging drugs and female sex is pro-arrhythmic.

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Generation of patient-specific induced pluripotent stem cell lines from one patient with Jervell and Lange-Nielsen syndrome, one with type 1 long QT syndrome and two healthy relatives

Stem Cell Research ◽

10.1016/j.scr.2018.07.016 ◽

2018 ◽

Vol 31 ◽

pp. 174-180 ◽

Cited By ~ 4

Author(s):

T.H. Kasai-Brunswick ◽

D. Silva dos Santos ◽

R.P. Ferreira ◽

D.S. Araujo ◽

G.M. Dias ◽

...

Keyword(s):

Stem Cell ◽

Long Qt Syndrome ◽

Pluripotent Stem Cell ◽

Induced Pluripotent Stem Cell ◽

Patient Specific ◽

Long Qt ◽

Qt Syndrome ◽

Stem Cell Lines ◽

Induced Pluripotent

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Studying KCNQ1 Mutation and Drug Response in Type 1 Long QT Syndrome Using Patient-Specific Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Methods in Molecular Biology - Potassium Channels ◽

10.1007/978-1-4939-7362-0_2 ◽

2017 ◽

pp. 7-28 ◽

Cited By ~ 1

Author(s):

Heming Wei ◽

Jianjun Wu ◽

Zhenfeng Liu

Keyword(s):

Stem Cell ◽

Long Qt Syndrome ◽

Pluripotent Stem Cell ◽

Drug Response ◽

Induced Pluripotent Stem Cell ◽

Patient Specific ◽

Long Qt ◽

Qt Syndrome ◽

Induced Pluripotent

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Abstract 142: Modeling Drug-Induced Long QT Syndrome with Patient-Specific Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Circulation Research ◽

10.1161/res.113.suppl_1.a142 ◽

2013 ◽

Vol 113 (suppl_1) ◽

Author(s):

Francesca Stillitano ◽

Ioannis Karakikes ◽

Chi-wai Kong ◽

Brett Martinelli ◽

Ronald Li ◽

...

Keyword(s):

Stem Cell ◽

Pluripotent Stem Cell ◽

Induced Pluripotent Stem Cell ◽

Patient Specific ◽

Dependent Manner ◽

Long Qt ◽

Drug Induced ◽

Qt Syndrome ◽

Induced Pluripotent ◽

Dose Dependent

Long QT syndrome (LQTS) is characterized by prolonged cardiac repolarization time and increased risk of ventricular arrhythmia. LQTS can be either inherited or induced notably after drugs intake. Mutations in genes encoding cardiac ion channels have been reported to underlie inherited LQTS. In contrast, drug-induced LQTS (diLQTS) most frequently arises from altered function of the hERG channel; the risk of developing diLQTS varies largely between subjects and most people who have life-threatening diLQTS have no known genetic risk factors. We investigated whether the susceptibility to develop diLQTS observed in vivo can be recapitulated in vitro using patient-specific induced pluripotent stem cell (iPSC) technology. We collected skin fibroblasts from ten subjects who developed significant diLQTS after administration of Sotalol and/or Erythromycin. Ten other individuals who displayed no changes in QT interval after administration of the same drugs, were selected. iPSC were generated by retroviral delivery of Oct4, Sox2, Nanog and Klf4 in 17 of the 20 individuals. We report preliminary results obtained from iPSC-derived cardiomyocytes (iPSC-CMs) of two subjects. All experiments were performed in a blinded fashion without knowledge of the associated clinical phenotype. Cardiac differentiation of iPSC resulted in the generation of spontaneously beating embryoid bodies. iPSC-CMs showed positive staining for TNNT2, ACTN2 and Cx43. Gene expression analysis confirmed the expression of NKX2.5, MLC2v, MYH6 and MYH7, and of the relevant KCNH2 gene. The two lines had similar basal electrophysiological properties as assessed by measurements of action potential (AP) by patch-clamp technique and extracellular field potentials (FP) using micro-electrode array (MEA). E4031, a classical HERG blocker, significantly prolonged the FP duration (FPD) in a dose-dependent manner in both lines (EC50: 30.19 and 51.57 respectively). When both Sotalol and Erythromicin were used, FPD was prolonged in one of the two samples in a dose-dependent manner (EC50Sotalol: 100; EC50Erythr: 9.64) while drug response was blunted in the other cell line. This study suggests that patient-specific iPSC can be used to model the functional abnormalities observed in acquired diLQTS.

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