In vitro modeling of ryanodine receptor 2 dysfunction using human induced pluripotent stem cells

Azra Fatima, Guoxing Xu, Kaifeng Shao, Symeon Papadopoulos, Martin Lehmann, Juan J Arnáiz-Cot, Angelo O Rosa, Filomain Nguemo, Matthias Matzkies, Sven Dittmann, Susannah L Stone, Matthias Linke, Ulrich Zechner, Vera Beyer, Hans Christian Hennies, Stephan Rosenkranz, Baerbel Klauke, Abdul S Parwani, Wilhelm Haverkamp, Gabriele Pfitzer & 6 others Martin Farr, Lars Cleemann, Martin Morad, Hendrik Milting, Juergen Hescheler, Tomo Saric

Research output: Contribution to journalArticle

118 Citations (Scopus)

Abstract

BACKGROUND/AIMS: Induced pluripotent stem (iPS) cells generated from accessible adult cells of patients with genetic diseases open unprecedented opportunities for exploring the pathophysiology of human diseases in vitro. Catecholaminergic polymorphic ventricular tachycardia type 1 (CPVT1) is an inherited cardiac disorder that is caused by mutations in the cardiac ryanodine receptor type 2 gene (RYR2) and is characterized by stress-induced ventricular arrhythmia that can lead to sudden cardiac death in young individuals. The aim of this study was to generate iPS cells from a patient with CPVT1 and determine whether iPS cell-derived cardiomyocytes carrying patient specific RYR2 mutation recapitulate the disease phenotype in vitro.

METHODS: iPS cells were derived from dermal fibroblasts of healthy donors and a patient with CPVT1 carrying the novel heterozygous autosomal dominant mutation p.F2483I in the RYR2. Functional properties of iPS cell derived-cardiomyocytes were analyzed by using whole-cell current and voltage clamp and calcium imaging techniques.

RESULTS: Patch-clamp recordings revealed arrhythmias and delayed afterdepolarizations (DADs) after catecholaminergic stimulation of CPVT1-iPS cell-derived cardiomyocytes. Calcium imaging studies showed that, compared to healthy cardiomyocytes, CPVT1-cardiomyocytes exhibit higher amplitudes and longer durations of spontaneous Ca(2+) release events at basal state. In addition, in CPVT1-cardiomyocytes the Ca(2+)-induced Ca(2+)-release events continued after repolarization and were abolished by increasing the cytosolic cAMP levels with forskolin.

CONCLUSION: This study demonstrates the suitability of iPS cells in modeling RYR2-related cardiac disorders in vitro and opens new opportunities for investigating the disease mechanism in vitro, developing new drugs, predicting their toxicity, and optimizing current treatment strategies.

Original languageEnglish
Pages (from-to)579-592
Number of pages14
JournalCellular Physiology and Biochemistry
Volume28
Issue number4
DOIs
Publication statusPublished - Dec 2011
Externally publishedYes

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Induced Pluripotent Stem Cells
Ryanodine Receptor Calcium Release Channel
Cardiac Myocytes
Mutation
Genes
Cardiac Arrhythmias
Calcium
Inborn Genetic Diseases
Sudden Cardiac Death
Colforsin
In Vitro Techniques
Polymorphic catecholergic ventricular tachycardia
Fibroblasts
Tissue Donors
Phenotype
Skin
Pharmaceutical Preparations

Cite this

Fatima, A., Xu, G., Shao, K., Papadopoulos, S., Lehmann, M., Arnáiz-Cot, J. J., ... Saric, T. (2011). In vitro modeling of ryanodine receptor 2 dysfunction using human induced pluripotent stem cells. Cellular Physiology and Biochemistry, 28(4), 579-592. https://doi.org/10.1159/000335753
Fatima, Azra ; Xu, Guoxing ; Shao, Kaifeng ; Papadopoulos, Symeon ; Lehmann, Martin ; Arnáiz-Cot, Juan J ; Rosa, Angelo O ; Nguemo, Filomain ; Matzkies, Matthias ; Dittmann, Sven ; Stone, Susannah L ; Linke, Matthias ; Zechner, Ulrich ; Beyer, Vera ; Hennies, Hans Christian ; Rosenkranz, Stephan ; Klauke, Baerbel ; Parwani, Abdul S ; Haverkamp, Wilhelm ; Pfitzer, Gabriele ; Farr, Martin ; Cleemann, Lars ; Morad, Martin ; Milting, Hendrik ; Hescheler, Juergen ; Saric, Tomo. / In vitro modeling of ryanodine receptor 2 dysfunction using human induced pluripotent stem cells. In: Cellular Physiology and Biochemistry. 2011 ; Vol. 28, No. 4. pp. 579-592.
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Fatima, A, Xu, G, Shao, K, Papadopoulos, S, Lehmann, M, Arnáiz-Cot, JJ, Rosa, AO, Nguemo, F, Matzkies, M, Dittmann, S, Stone, SL, Linke, M, Zechner, U, Beyer, V, Hennies, HC, Rosenkranz, S, Klauke, B, Parwani, AS, Haverkamp, W, Pfitzer, G, Farr, M, Cleemann, L, Morad, M, Milting, H, Hescheler, J & Saric, T 2011, 'In vitro modeling of ryanodine receptor 2 dysfunction using human induced pluripotent stem cells', Cellular Physiology and Biochemistry, vol. 28, no. 4, pp. 579-592. https://doi.org/10.1159/000335753

In vitro modeling of ryanodine receptor 2 dysfunction using human induced pluripotent stem cells. / Fatima, Azra; Xu, Guoxing; Shao, Kaifeng; Papadopoulos, Symeon; Lehmann, Martin; Arnáiz-Cot, Juan J; Rosa, Angelo O; Nguemo, Filomain; Matzkies, Matthias; Dittmann, Sven; Stone, Susannah L; Linke, Matthias; Zechner, Ulrich; Beyer, Vera; Hennies, Hans Christian; Rosenkranz, Stephan; Klauke, Baerbel; Parwani, Abdul S; Haverkamp, Wilhelm; Pfitzer, Gabriele; Farr, Martin; Cleemann, Lars; Morad, Martin; Milting, Hendrik; Hescheler, Juergen; Saric, Tomo.

In: Cellular Physiology and Biochemistry, Vol. 28, No. 4, 12.2011, p. 579-592.

Research output: Contribution to journalArticle

TY - JOUR

T1 - In vitro modeling of ryanodine receptor 2 dysfunction using human induced pluripotent stem cells

AU - Fatima, Azra

AU - Xu, Guoxing

AU - Shao, Kaifeng

AU - Papadopoulos, Symeon

AU - Lehmann, Martin

AU - Arnáiz-Cot, Juan J

AU - Rosa, Angelo O

AU - Nguemo, Filomain

AU - Matzkies, Matthias

AU - Dittmann, Sven

AU - Stone, Susannah L

AU - Linke, Matthias

AU - Zechner, Ulrich

AU - Beyer, Vera

AU - Hennies, Hans Christian

AU - Rosenkranz, Stephan

AU - Klauke, Baerbel

AU - Parwani, Abdul S

AU - Haverkamp, Wilhelm

AU - Pfitzer, Gabriele

AU - Farr, Martin

AU - Cleemann, Lars

AU - Morad, Martin

AU - Milting, Hendrik

AU - Hescheler, Juergen

AU - Saric, Tomo

N1 - Copyright © 2011 S. Karger AG, Basel.

PY - 2011/12

Y1 - 2011/12

N2 - BACKGROUND/AIMS: Induced pluripotent stem (iPS) cells generated from accessible adult cells of patients with genetic diseases open unprecedented opportunities for exploring the pathophysiology of human diseases in vitro. Catecholaminergic polymorphic ventricular tachycardia type 1 (CPVT1) is an inherited cardiac disorder that is caused by mutations in the cardiac ryanodine receptor type 2 gene (RYR2) and is characterized by stress-induced ventricular arrhythmia that can lead to sudden cardiac death in young individuals. The aim of this study was to generate iPS cells from a patient with CPVT1 and determine whether iPS cell-derived cardiomyocytes carrying patient specific RYR2 mutation recapitulate the disease phenotype in vitro.METHODS: iPS cells were derived from dermal fibroblasts of healthy donors and a patient with CPVT1 carrying the novel heterozygous autosomal dominant mutation p.F2483I in the RYR2. Functional properties of iPS cell derived-cardiomyocytes were analyzed by using whole-cell current and voltage clamp and calcium imaging techniques.RESULTS: Patch-clamp recordings revealed arrhythmias and delayed afterdepolarizations (DADs) after catecholaminergic stimulation of CPVT1-iPS cell-derived cardiomyocytes. Calcium imaging studies showed that, compared to healthy cardiomyocytes, CPVT1-cardiomyocytes exhibit higher amplitudes and longer durations of spontaneous Ca(2+) release events at basal state. In addition, in CPVT1-cardiomyocytes the Ca(2+)-induced Ca(2+)-release events continued after repolarization and were abolished by increasing the cytosolic cAMP levels with forskolin.CONCLUSION: This study demonstrates the suitability of iPS cells in modeling RYR2-related cardiac disorders in vitro and opens new opportunities for investigating the disease mechanism in vitro, developing new drugs, predicting their toxicity, and optimizing current treatment strategies.

AB - BACKGROUND/AIMS: Induced pluripotent stem (iPS) cells generated from accessible adult cells of patients with genetic diseases open unprecedented opportunities for exploring the pathophysiology of human diseases in vitro. Catecholaminergic polymorphic ventricular tachycardia type 1 (CPVT1) is an inherited cardiac disorder that is caused by mutations in the cardiac ryanodine receptor type 2 gene (RYR2) and is characterized by stress-induced ventricular arrhythmia that can lead to sudden cardiac death in young individuals. The aim of this study was to generate iPS cells from a patient with CPVT1 and determine whether iPS cell-derived cardiomyocytes carrying patient specific RYR2 mutation recapitulate the disease phenotype in vitro.METHODS: iPS cells were derived from dermal fibroblasts of healthy donors and a patient with CPVT1 carrying the novel heterozygous autosomal dominant mutation p.F2483I in the RYR2. Functional properties of iPS cell derived-cardiomyocytes were analyzed by using whole-cell current and voltage clamp and calcium imaging techniques.RESULTS: Patch-clamp recordings revealed arrhythmias and delayed afterdepolarizations (DADs) after catecholaminergic stimulation of CPVT1-iPS cell-derived cardiomyocytes. Calcium imaging studies showed that, compared to healthy cardiomyocytes, CPVT1-cardiomyocytes exhibit higher amplitudes and longer durations of spontaneous Ca(2+) release events at basal state. In addition, in CPVT1-cardiomyocytes the Ca(2+)-induced Ca(2+)-release events continued after repolarization and were abolished by increasing the cytosolic cAMP levels with forskolin.CONCLUSION: This study demonstrates the suitability of iPS cells in modeling RYR2-related cardiac disorders in vitro and opens new opportunities for investigating the disease mechanism in vitro, developing new drugs, predicting their toxicity, and optimizing current treatment strategies.

KW - Action Potentials

KW - Calcium

KW - Catecholamines

KW - Cell Differentiation

KW - Colforsin

KW - Cyclic AMP

KW - Electrocardiography

KW - Heterozygote

KW - Humans

KW - Induced Pluripotent Stem Cells

KW - Karyotyping

KW - Models, Biological

KW - Mutation

KW - Myocytes, Cardiac

KW - Patch-Clamp Techniques

KW - Phenotype

KW - Ryanodine Receptor Calcium Release Channel

KW - Tachycardia, Ventricular

KW - Journal Article

KW - Research Support, N.I.H., Extramural

KW - Research Support, Non-U.S. Gov't

U2 - 10.1159/000335753

DO - 10.1159/000335753

M3 - Article

VL - 28

SP - 579

EP - 592

JO - Cellular Physiology and Biochemistry

JF - Cellular Physiology and Biochemistry

SN - 1015-8987

IS - 4

ER -