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Overexpression of KCNJ2 enhances maturation of human-induced pluripotent stem cell-derived cardiomyocytes
BACKGROUND: Although human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) are a promising cell resource for cardiovascular research, these cells exhibit an immature phenotype that hampers their potential applications. The inwardly rectifying potassium channel K(ir)2.1, encoded by th...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10105952/ https://www.ncbi.nlm.nih.gov/pubmed/37061738 http://dx.doi.org/10.1186/s13287-023-03312-9 |
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author | Zhou, Jingjun Cui, Baiping Wang, Xiaochen Wang, Hongkun Zheng, Junnan Guo, Fengfeng Sun, Yaxun Fan, Hangping Shen, Jiaxi Su, Jun Wang, Jue Zhao, Haige Tang, Yiquan Gong, Tingyu Sun, Ning Liang, Ping |
author_facet | Zhou, Jingjun Cui, Baiping Wang, Xiaochen Wang, Hongkun Zheng, Junnan Guo, Fengfeng Sun, Yaxun Fan, Hangping Shen, Jiaxi Su, Jun Wang, Jue Zhao, Haige Tang, Yiquan Gong, Tingyu Sun, Ning Liang, Ping |
author_sort | Zhou, Jingjun |
collection | PubMed |
description | BACKGROUND: Although human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) are a promising cell resource for cardiovascular research, these cells exhibit an immature phenotype that hampers their potential applications. The inwardly rectifying potassium channel K(ir)2.1, encoded by the KCNJ2 gene, has been thought as an important target for promoting electrical maturation of iPSC-CMs. However, a comprehensive characterization of morphological and functional changes in iPSC-CMs overexpressing KCNJ2 (KCNJ2 OE) is still lacking. METHODS: iPSC-CMs were generated using a 2D in vitro monolayer differentiation protocol. Human KCNJ2 construct with green fluorescent protein (GFP) tag was created and overexpressed in iPSC-CMs via lentiviral transduction. The mixture of iPSC-CMs and mesenchymal cells was cocultured with decellularized natural heart matrix for generation of 3D human engineered heart tissues (EHTs). RESULTS: We showed that mRNA expression level of KCNJ2 in iPSC-CMs was dramatically lower than that in human left ventricular tissues. KCNJ2 OE iPSC-CMs yielded significantly increased protein expression of K(ir)2.1 and current density of K(ir)2.1-encoded I(K1). The larger I(K1) linked to a quiescent phenotype that required pacing to elicit action potentials in KCNJ2 OE iPSC-CMs, which can be reversed by I(K1) blocker BaCl(2). KCNJ2 OE also led to significantly hyperpolarized maximal diastolic potential (MDP), shortened action potential duration (APD) and increased maximal upstroke velocity. The enhanced electrophysiological maturation in KCNJ2 OE iPSC-CMs was accompanied by improvements in Ca(2+) signaling, mitochondrial energy metabolism and transcriptomic profile. Notably, KCNJ2 OE iPSC-CMs exhibited enlarged cell size and more elongated and stretched shape, indicating a morphological phenotype toward structural maturation. Drug testing using hERG blocker E-4031 revealed that a more stable MDP in KCNJ2 OE iPSC-CMs allowed for obtaining significant drug response of APD prolongation in a concentration-dependent manner. Moreover, KCNJ2 OE iPSC-CMs formed more mature human EHTs with better tissue structure and cell junction. CONCLUSIONS: Overexpression of KCNJ2 can robustly enhance maturation of iPSC-CMs in electrophysiology, Ca(2+) signaling, metabolism, transcriptomic profile, cardiomyocyte structure and tissue engineering, thus providing more accurate cellular model for elucidating cellular and molecular mechanisms of cardiovascular diseases, screening drug-induced cardiotoxicity, and developing personalized and precision cardiovascular medicine. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-023-03312-9. |
format | Online Article Text |
id | pubmed-10105952 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-101059522023-04-17 Overexpression of KCNJ2 enhances maturation of human-induced pluripotent stem cell-derived cardiomyocytes Zhou, Jingjun Cui, Baiping Wang, Xiaochen Wang, Hongkun Zheng, Junnan Guo, Fengfeng Sun, Yaxun Fan, Hangping Shen, Jiaxi Su, Jun Wang, Jue Zhao, Haige Tang, Yiquan Gong, Tingyu Sun, Ning Liang, Ping Stem Cell Res Ther Research BACKGROUND: Although human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) are a promising cell resource for cardiovascular research, these cells exhibit an immature phenotype that hampers their potential applications. The inwardly rectifying potassium channel K(ir)2.1, encoded by the KCNJ2 gene, has been thought as an important target for promoting electrical maturation of iPSC-CMs. However, a comprehensive characterization of morphological and functional changes in iPSC-CMs overexpressing KCNJ2 (KCNJ2 OE) is still lacking. METHODS: iPSC-CMs were generated using a 2D in vitro monolayer differentiation protocol. Human KCNJ2 construct with green fluorescent protein (GFP) tag was created and overexpressed in iPSC-CMs via lentiviral transduction. The mixture of iPSC-CMs and mesenchymal cells was cocultured with decellularized natural heart matrix for generation of 3D human engineered heart tissues (EHTs). RESULTS: We showed that mRNA expression level of KCNJ2 in iPSC-CMs was dramatically lower than that in human left ventricular tissues. KCNJ2 OE iPSC-CMs yielded significantly increased protein expression of K(ir)2.1 and current density of K(ir)2.1-encoded I(K1). The larger I(K1) linked to a quiescent phenotype that required pacing to elicit action potentials in KCNJ2 OE iPSC-CMs, which can be reversed by I(K1) blocker BaCl(2). KCNJ2 OE also led to significantly hyperpolarized maximal diastolic potential (MDP), shortened action potential duration (APD) and increased maximal upstroke velocity. The enhanced electrophysiological maturation in KCNJ2 OE iPSC-CMs was accompanied by improvements in Ca(2+) signaling, mitochondrial energy metabolism and transcriptomic profile. Notably, KCNJ2 OE iPSC-CMs exhibited enlarged cell size and more elongated and stretched shape, indicating a morphological phenotype toward structural maturation. Drug testing using hERG blocker E-4031 revealed that a more stable MDP in KCNJ2 OE iPSC-CMs allowed for obtaining significant drug response of APD prolongation in a concentration-dependent manner. Moreover, KCNJ2 OE iPSC-CMs formed more mature human EHTs with better tissue structure and cell junction. CONCLUSIONS: Overexpression of KCNJ2 can robustly enhance maturation of iPSC-CMs in electrophysiology, Ca(2+) signaling, metabolism, transcriptomic profile, cardiomyocyte structure and tissue engineering, thus providing more accurate cellular model for elucidating cellular and molecular mechanisms of cardiovascular diseases, screening drug-induced cardiotoxicity, and developing personalized and precision cardiovascular medicine. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-023-03312-9. BioMed Central 2023-04-15 /pmc/articles/PMC10105952/ /pubmed/37061738 http://dx.doi.org/10.1186/s13287-023-03312-9 Text en © The Author(s) 2023 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/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Zhou, Jingjun Cui, Baiping Wang, Xiaochen Wang, Hongkun Zheng, Junnan Guo, Fengfeng Sun, Yaxun Fan, Hangping Shen, Jiaxi Su, Jun Wang, Jue Zhao, Haige Tang, Yiquan Gong, Tingyu Sun, Ning Liang, Ping Overexpression of KCNJ2 enhances maturation of human-induced pluripotent stem cell-derived cardiomyocytes |
title | Overexpression of KCNJ2 enhances maturation of human-induced pluripotent stem cell-derived cardiomyocytes |
title_full | Overexpression of KCNJ2 enhances maturation of human-induced pluripotent stem cell-derived cardiomyocytes |
title_fullStr | Overexpression of KCNJ2 enhances maturation of human-induced pluripotent stem cell-derived cardiomyocytes |
title_full_unstemmed | Overexpression of KCNJ2 enhances maturation of human-induced pluripotent stem cell-derived cardiomyocytes |
title_short | Overexpression of KCNJ2 enhances maturation of human-induced pluripotent stem cell-derived cardiomyocytes |
title_sort | overexpression of kcnj2 enhances maturation of human-induced pluripotent stem cell-derived cardiomyocytes |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10105952/ https://www.ncbi.nlm.nih.gov/pubmed/37061738 http://dx.doi.org/10.1186/s13287-023-03312-9 |
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