Integrins Increase Sarcoplasmic Reticulum Activity for Excitation—Contraction Coupling in Human Stem Cell-Derived Cardiomyocytes
Engagement of the sarcoplasmic reticulum (SR) Ca<sup>2+</sup> stores for excitation–contraction (EC)-coupling is a fundamental feature of cardiac muscle cells. Extracellular matrix (ECM) proteins that form the extracellular scaffolding supporting cardiac contractile activity are thought...
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MDPI AG
2022-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/18/10940 |
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| author | Brian X. Wang Christopher Kane Laura Nicastro Oisín King Worrapong Kit-Anan Barrett Downing Graziano Deidda Liam S. Couch Christian Pinali Anna Mitraki Kenneth T. MacLeod Cesare M. Terracciano |
| author_facet | Brian X. Wang Christopher Kane Laura Nicastro Oisín King Worrapong Kit-Anan Barrett Downing Graziano Deidda Liam S. Couch Christian Pinali Anna Mitraki Kenneth T. MacLeod Cesare M. Terracciano |
| author_sort | Brian X. Wang |
| collection | DOAJ |
| description | Engagement of the sarcoplasmic reticulum (SR) Ca<sup>2+</sup> stores for excitation–contraction (EC)-coupling is a fundamental feature of cardiac muscle cells. Extracellular matrix (ECM) proteins that form the extracellular scaffolding supporting cardiac contractile activity are thought to play an integral role in the modulation of EC-coupling. At baseline, human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) show poor utilisation of SR Ca<sup>2+</sup> stores, leading to inefficient EC-coupling, like developing or human CMs in cardiac diseases such as heart failure. We hypothesised that integrin ligand–receptor interactions between ECM proteins and CMs recruit the SR to Ca<sup>2+</sup> cycling during EC-coupling. hiPSC-CM monolayers were cultured on fibronectin-coated glass before 24 h treatment with fibril-forming peptides containing the integrin-binding tripeptide sequence arginine–glycine–aspartic acid (2 mM). Micropipette application of 40 mM caffeine in standard or Na<sup>+</sup>/Ca<sup>2+</sup>-free Tyrode’s solutions was used to assess the Ca<sup>2+</sup> removal mechanisms. Microelectrode recordings were conducted to analyse action potentials in current-clamp. Confocal images of labelled hiPSC-CMs were analysed to investigate hiPSC-CM morphology and ultrastructural arrangements in Ca<sup>2+</sup> release units. This study demonstrates that peptides containing the integrin-binding sequence arginine–glycine–aspartic acid (1) abbreviate hiPSC-CM Ca<sup>2+</sup> transient and action potential duration, (2) increase co-localisation between L-type Ca<sup>2+</sup> channels and ryanodine receptors involved in EC-coupling, and (3) increase the rate of SR-mediated Ca<sup>2+</sup> cycling. We conclude that integrin-binding peptides induce recruitment of the SR for Ca<sup>2+</sup> cycling in EC-coupling through functional and structural improvements and demonstrate the importance of the ECM in modulating cardiomyocyte function in physiology. |
| first_indexed | 2024-03-09T23:42:06Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T23:42:06Z |
| publishDate | 2022-09-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-a0c335a772f34dd3b77998664e203c382023-11-23T16:50:51ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-09-0123181094010.3390/ijms231810940Integrins Increase Sarcoplasmic Reticulum Activity for Excitation—Contraction Coupling in Human Stem Cell-Derived CardiomyocytesBrian X. Wang0Christopher Kane1Laura Nicastro2Oisín King3Worrapong Kit-Anan4Barrett Downing5Graziano Deidda6Liam S. Couch7Christian Pinali8Anna Mitraki9Kenneth T. MacLeod10Cesare M. Terracciano11National Heart & Lung Institute, Imperial College London, London SW7 2AZ, UKNational Heart & Lung Institute, Imperial College London, London SW7 2AZ, UKNational Heart & Lung Institute, Imperial College London, London SW7 2AZ, UKNational Heart & Lung Institute, Imperial College London, London SW7 2AZ, UKNational Heart & Lung Institute, Imperial College London, London SW7 2AZ, UKNational Heart & Lung Institute, Imperial College London, London SW7 2AZ, UKInstitute of Electronic Structure and Laser (IESL), Foundation for Research and Technology−Hellas (FORTH), 700 13 Heraklion, GreeceNational Heart & Lung Institute, Imperial College London, London SW7 2AZ, UKDivision of Cardiovascular Sciences, University of Manchester, Manchester M13 9NT, UKInstitute of Electronic Structure and Laser (IESL), Foundation for Research and Technology−Hellas (FORTH), 700 13 Heraklion, GreeceNational Heart & Lung Institute, Imperial College London, London SW7 2AZ, UKNational Heart & Lung Institute, Imperial College London, London SW7 2AZ, UKEngagement of the sarcoplasmic reticulum (SR) Ca<sup>2+</sup> stores for excitation–contraction (EC)-coupling is a fundamental feature of cardiac muscle cells. Extracellular matrix (ECM) proteins that form the extracellular scaffolding supporting cardiac contractile activity are thought to play an integral role in the modulation of EC-coupling. At baseline, human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) show poor utilisation of SR Ca<sup>2+</sup> stores, leading to inefficient EC-coupling, like developing or human CMs in cardiac diseases such as heart failure. We hypothesised that integrin ligand–receptor interactions between ECM proteins and CMs recruit the SR to Ca<sup>2+</sup> cycling during EC-coupling. hiPSC-CM monolayers were cultured on fibronectin-coated glass before 24 h treatment with fibril-forming peptides containing the integrin-binding tripeptide sequence arginine–glycine–aspartic acid (2 mM). Micropipette application of 40 mM caffeine in standard or Na<sup>+</sup>/Ca<sup>2+</sup>-free Tyrode’s solutions was used to assess the Ca<sup>2+</sup> removal mechanisms. Microelectrode recordings were conducted to analyse action potentials in current-clamp. Confocal images of labelled hiPSC-CMs were analysed to investigate hiPSC-CM morphology and ultrastructural arrangements in Ca<sup>2+</sup> release units. This study demonstrates that peptides containing the integrin-binding sequence arginine–glycine–aspartic acid (1) abbreviate hiPSC-CM Ca<sup>2+</sup> transient and action potential duration, (2) increase co-localisation between L-type Ca<sup>2+</sup> channels and ryanodine receptors involved in EC-coupling, and (3) increase the rate of SR-mediated Ca<sup>2+</sup> cycling. We conclude that integrin-binding peptides induce recruitment of the SR for Ca<sup>2+</sup> cycling in EC-coupling through functional and structural improvements and demonstrate the importance of the ECM in modulating cardiomyocyte function in physiology.https://www.mdpi.com/1422-0067/23/18/10940stem cellintegrinCa<sup>2+</sup> cyclingcardiomyocytefibrosissarcoplasmic reticulum |
| spellingShingle | Brian X. Wang Christopher Kane Laura Nicastro Oisín King Worrapong Kit-Anan Barrett Downing Graziano Deidda Liam S. Couch Christian Pinali Anna Mitraki Kenneth T. MacLeod Cesare M. Terracciano Integrins Increase Sarcoplasmic Reticulum Activity for Excitation—Contraction Coupling in Human Stem Cell-Derived Cardiomyocytes International Journal of Molecular Sciences stem cell integrin Ca<sup>2+</sup> cycling cardiomyocyte fibrosis sarcoplasmic reticulum |
| title | Integrins Increase Sarcoplasmic Reticulum Activity for Excitation—Contraction Coupling in Human Stem Cell-Derived Cardiomyocytes |
| title_full | Integrins Increase Sarcoplasmic Reticulum Activity for Excitation—Contraction Coupling in Human Stem Cell-Derived Cardiomyocytes |
| title_fullStr | Integrins Increase Sarcoplasmic Reticulum Activity for Excitation—Contraction Coupling in Human Stem Cell-Derived Cardiomyocytes |
| title_full_unstemmed | Integrins Increase Sarcoplasmic Reticulum Activity for Excitation—Contraction Coupling in Human Stem Cell-Derived Cardiomyocytes |
| title_short | Integrins Increase Sarcoplasmic Reticulum Activity for Excitation—Contraction Coupling in Human Stem Cell-Derived Cardiomyocytes |
| title_sort | integrins increase sarcoplasmic reticulum activity for excitation contraction coupling in human stem cell derived cardiomyocytes |
| topic | stem cell integrin Ca<sup>2+</sup> cycling cardiomyocyte fibrosis sarcoplasmic reticulum |
| url | https://www.mdpi.com/1422-0067/23/18/10940 |
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