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Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual I(K1)

Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) are widely used in studying basic mechanisms of cardiac arrhythmias that are caused by ion channelopathies. Unfortunately, the action potential profile of hiPSC-CMs—and consequently the profile of individual membrane currents act...

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Autores principales: Meijer van Putten, Rosalie M. E., Mengarelli, Isabella, Guan, Kaomei, Zegers, Jan G., van Ginneken, Antoni C. G., Verkerk, Arie O., Wilders, Ronald
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4315032/
https://www.ncbi.nlm.nih.gov/pubmed/25691870
http://dx.doi.org/10.3389/fphys.2015.00007
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author Meijer van Putten, Rosalie M. E.
Mengarelli, Isabella
Guan, Kaomei
Zegers, Jan G.
van Ginneken, Antoni C. G.
Verkerk, Arie O.
Wilders, Ronald
author_facet Meijer van Putten, Rosalie M. E.
Mengarelli, Isabella
Guan, Kaomei
Zegers, Jan G.
van Ginneken, Antoni C. G.
Verkerk, Arie O.
Wilders, Ronald
author_sort Meijer van Putten, Rosalie M. E.
collection PubMed
description Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) are widely used in studying basic mechanisms of cardiac arrhythmias that are caused by ion channelopathies. Unfortunately, the action potential profile of hiPSC-CMs—and consequently the profile of individual membrane currents active during that action potential—differs substantially from that of native human cardiomyocytes, largely due to almost negligible expression of the inward rectifier potassium current (I(K1)). In the present study, we attempted to “normalize” the action potential profile of our hiPSC-CMs by inserting a voltage dependent in silico I(K1) into our hiPSC-CMs, using the dynamic clamp configuration of the patch clamp technique. Recordings were made from single hiPSC-CMs, using the perforated patch clamp technique at physiological temperature. We assessed three different models of I(K1), with different degrees of inward rectification, and systematically varied the magnitude of the inserted I(K1). Also, we modified the inserted I(K1) in order to assess the effects of loss- and gain-of-function mutations in the KCNJ2 gene, which encodes the Kir2.1 protein that is primarily responsible for the I(K1) channel in human ventricle. For our experiments, we selected spontaneously beating hiPSC-CMs, with negligible I(K1) as demonstrated in separate voltage clamp experiments, which were paced at 1 Hz. Upon addition of in silico I(K1) with a peak outward density of 4–6 pA/pF, these hiPSC-CMs showed a ventricular-like action potential morphology with a stable resting membrane potential near −80 mV and a maximum upstroke velocity >150 V/s (n = 9). Proarrhythmic action potential changes were observed upon injection of both loss-of-function and gain-of-function I(K1), as associated with Andersen–Tawil syndrome type 1 and short QT syndrome type 3, respectively (n = 6). We conclude that injection of in silico I(K1) makes the hiPSC-CM a more reliable model for investigating mechanisms underlying cardiac arrhythmias.
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spelling pubmed-43150322015-02-17 Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual I(K1) Meijer van Putten, Rosalie M. E. Mengarelli, Isabella Guan, Kaomei Zegers, Jan G. van Ginneken, Antoni C. G. Verkerk, Arie O. Wilders, Ronald Front Physiol Physiology Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) are widely used in studying basic mechanisms of cardiac arrhythmias that are caused by ion channelopathies. Unfortunately, the action potential profile of hiPSC-CMs—and consequently the profile of individual membrane currents active during that action potential—differs substantially from that of native human cardiomyocytes, largely due to almost negligible expression of the inward rectifier potassium current (I(K1)). In the present study, we attempted to “normalize” the action potential profile of our hiPSC-CMs by inserting a voltage dependent in silico I(K1) into our hiPSC-CMs, using the dynamic clamp configuration of the patch clamp technique. Recordings were made from single hiPSC-CMs, using the perforated patch clamp technique at physiological temperature. We assessed three different models of I(K1), with different degrees of inward rectification, and systematically varied the magnitude of the inserted I(K1). Also, we modified the inserted I(K1) in order to assess the effects of loss- and gain-of-function mutations in the KCNJ2 gene, which encodes the Kir2.1 protein that is primarily responsible for the I(K1) channel in human ventricle. For our experiments, we selected spontaneously beating hiPSC-CMs, with negligible I(K1) as demonstrated in separate voltage clamp experiments, which were paced at 1 Hz. Upon addition of in silico I(K1) with a peak outward density of 4–6 pA/pF, these hiPSC-CMs showed a ventricular-like action potential morphology with a stable resting membrane potential near −80 mV and a maximum upstroke velocity >150 V/s (n = 9). Proarrhythmic action potential changes were observed upon injection of both loss-of-function and gain-of-function I(K1), as associated with Andersen–Tawil syndrome type 1 and short QT syndrome type 3, respectively (n = 6). We conclude that injection of in silico I(K1) makes the hiPSC-CM a more reliable model for investigating mechanisms underlying cardiac arrhythmias. Frontiers Media S.A. 2015-02-03 /pmc/articles/PMC4315032/ /pubmed/25691870 http://dx.doi.org/10.3389/fphys.2015.00007 Text en Copyright © 2015 Meijer van Putten, Mengarelli, Guan, Zegers, van Ginneken, Verkerk and Wilders. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Physiology
Meijer van Putten, Rosalie M. E.
Mengarelli, Isabella
Guan, Kaomei
Zegers, Jan G.
van Ginneken, Antoni C. G.
Verkerk, Arie O.
Wilders, Ronald
Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual I(K1)
title Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual I(K1)
title_full Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual I(K1)
title_fullStr Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual I(K1)
title_full_unstemmed Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual I(K1)
title_short Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual I(K1)
title_sort ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual i(k1)
topic Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4315032/
https://www.ncbi.nlm.nih.gov/pubmed/25691870
http://dx.doi.org/10.3389/fphys.2015.00007
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