Increased cytosolic calcium buffering contributes to a cellular arrhythmogenic substrate in iPSC-cardiomyocytes from patients with dilated cardiomyopathy
Dilated cardiomyopathy (DCM) is a major risk factor for heart failure and is associated with the development of life-threatening cardiac arrhythmias. Using a patient-specific induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM) model harbouring a mutation in cardiac troponin T (R173W), we a...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Springer Berlin Heidelberg
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9061684/ https://www.ncbi.nlm.nih.gov/pubmed/35499658 http://dx.doi.org/10.1007/s00395-022-00912-z |
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author | Jung, Philipp Seibertz, Fitzwilliam Fakuade, Funsho E. Ignatyeva, Nadezda Sampathkumar, Shrivatsan Ritter, Melanie Li, Housen Mason, Fleur E. Ebert, Antje Voigt, Niels |
author_facet | Jung, Philipp Seibertz, Fitzwilliam Fakuade, Funsho E. Ignatyeva, Nadezda Sampathkumar, Shrivatsan Ritter, Melanie Li, Housen Mason, Fleur E. Ebert, Antje Voigt, Niels |
author_sort | Jung, Philipp |
collection | PubMed |
description | Dilated cardiomyopathy (DCM) is a major risk factor for heart failure and is associated with the development of life-threatening cardiac arrhythmias. Using a patient-specific induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM) model harbouring a mutation in cardiac troponin T (R173W), we aim to examine the cellular basis of arrhythmogenesis in DCM patients with this mutation. iPSC from control (Ctrl) and DCM-TnT-R173W donors from the same family were differentiated into iPSC-CM and analysed through optical action potential (AP) recordings, simultaneous measurement of cytosolic calcium concentration ([Ca(2+)](i)) and membrane currents and separately assayed using field stimulation to detect the threshold for AP- and [Ca(2+)](i)-alternans development. AP duration was unaltered in TnT-R173W iPSC-CM. Nevertheless, TnT-R173W iPSC-CM showed a strikingly low stimulation threshold for AP- and [Ca(2+)](i)-alternans. Myofilaments are known to play a role as intracellular Ca(2+) buffers and here we show increased Ca(2+) affinity of intracellular buffers in TnT-R173W cells, indicating increased myofilament sensitivity to Ca(2+). Similarly, EMD57033, a myofilament Ca(2+) sensitiser, replicated the abnormal [Ca(2+)](i) dynamics observed in TnT-R173W samples and lowered the threshold for alternans development. In contrast, application of a Ca(2+) desensitiser (blebbistatin) to TnT-R173W iPSC-CM was able to phenotypically rescue Ca(2+) dynamics, normalising Ca(2+) transient profile and minimising the occurrence of Ca(2+) alternans at physiological frequencies. This finding suggests that increased Ca(2+) buffering likely plays a major arrhythmogenic role in patients with DCM, specifically in those with mutations in cardiac troponin T. In addition, we propose that modulation of myofilament Ca(2+) sensitivity could be an effective anti-arrhythmic target for pharmacological management of this disease. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00395-022-00912-z. |
format | Online Article Text |
id | pubmed-9061684 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-90616842022-05-07 Increased cytosolic calcium buffering contributes to a cellular arrhythmogenic substrate in iPSC-cardiomyocytes from patients with dilated cardiomyopathy Jung, Philipp Seibertz, Fitzwilliam Fakuade, Funsho E. Ignatyeva, Nadezda Sampathkumar, Shrivatsan Ritter, Melanie Li, Housen Mason, Fleur E. Ebert, Antje Voigt, Niels Basic Res Cardiol Original Contribution Dilated cardiomyopathy (DCM) is a major risk factor for heart failure and is associated with the development of life-threatening cardiac arrhythmias. Using a patient-specific induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM) model harbouring a mutation in cardiac troponin T (R173W), we aim to examine the cellular basis of arrhythmogenesis in DCM patients with this mutation. iPSC from control (Ctrl) and DCM-TnT-R173W donors from the same family were differentiated into iPSC-CM and analysed through optical action potential (AP) recordings, simultaneous measurement of cytosolic calcium concentration ([Ca(2+)](i)) and membrane currents and separately assayed using field stimulation to detect the threshold for AP- and [Ca(2+)](i)-alternans development. AP duration was unaltered in TnT-R173W iPSC-CM. Nevertheless, TnT-R173W iPSC-CM showed a strikingly low stimulation threshold for AP- and [Ca(2+)](i)-alternans. Myofilaments are known to play a role as intracellular Ca(2+) buffers and here we show increased Ca(2+) affinity of intracellular buffers in TnT-R173W cells, indicating increased myofilament sensitivity to Ca(2+). Similarly, EMD57033, a myofilament Ca(2+) sensitiser, replicated the abnormal [Ca(2+)](i) dynamics observed in TnT-R173W samples and lowered the threshold for alternans development. In contrast, application of a Ca(2+) desensitiser (blebbistatin) to TnT-R173W iPSC-CM was able to phenotypically rescue Ca(2+) dynamics, normalising Ca(2+) transient profile and minimising the occurrence of Ca(2+) alternans at physiological frequencies. This finding suggests that increased Ca(2+) buffering likely plays a major arrhythmogenic role in patients with DCM, specifically in those with mutations in cardiac troponin T. In addition, we propose that modulation of myofilament Ca(2+) sensitivity could be an effective anti-arrhythmic target for pharmacological management of this disease. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00395-022-00912-z. Springer Berlin Heidelberg 2022-05-02 2022 /pmc/articles/PMC9061684/ /pubmed/35499658 http://dx.doi.org/10.1007/s00395-022-00912-z Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Original Contribution Jung, Philipp Seibertz, Fitzwilliam Fakuade, Funsho E. Ignatyeva, Nadezda Sampathkumar, Shrivatsan Ritter, Melanie Li, Housen Mason, Fleur E. Ebert, Antje Voigt, Niels Increased cytosolic calcium buffering contributes to a cellular arrhythmogenic substrate in iPSC-cardiomyocytes from patients with dilated cardiomyopathy |
title | Increased cytosolic calcium buffering contributes to a cellular arrhythmogenic substrate in iPSC-cardiomyocytes from patients with dilated cardiomyopathy |
title_full | Increased cytosolic calcium buffering contributes to a cellular arrhythmogenic substrate in iPSC-cardiomyocytes from patients with dilated cardiomyopathy |
title_fullStr | Increased cytosolic calcium buffering contributes to a cellular arrhythmogenic substrate in iPSC-cardiomyocytes from patients with dilated cardiomyopathy |
title_full_unstemmed | Increased cytosolic calcium buffering contributes to a cellular arrhythmogenic substrate in iPSC-cardiomyocytes from patients with dilated cardiomyopathy |
title_short | Increased cytosolic calcium buffering contributes to a cellular arrhythmogenic substrate in iPSC-cardiomyocytes from patients with dilated cardiomyopathy |
title_sort | increased cytosolic calcium buffering contributes to a cellular arrhythmogenic substrate in ipsc-cardiomyocytes from patients with dilated cardiomyopathy |
topic | Original Contribution |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9061684/ https://www.ncbi.nlm.nih.gov/pubmed/35499658 http://dx.doi.org/10.1007/s00395-022-00912-z |
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