Orai1–STIM1 Regulates Increased Ca2+ Mobilization, Leading to Contractile Duchenne Muscular Dystrophy Phenotypes in Patient-Derived Induced Pluripotent Stem Cells

Ca2+ overload is one of the factors leading to Duchenne muscular dystrophy (DMD) pathogenesis. However, the molecular targets of dystrophin deficiency-dependent Ca2+ overload and the correlation between Ca2+ overload and contractil...

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Autores principales:	Tomoya Uchimura, Hidetoshi Sakurai
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	iPSC skeletal muscle Ca<sup>2+</sup> overload store-operated Ca<sup>2+</sup> channel STIM1-Orai1 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/d09301e338eb479ca80f3f40bd7f4477
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spelling	oai:doaj.org-article:d09301e338eb479ca80f3f40bd7f44772021-11-25T16:49:22ZOrai1–STIM1 Regulates Increased Ca<sup>2+</sup> Mobilization, Leading to Contractile Duchenne Muscular Dystrophy Phenotypes in Patient-Derived Induced Pluripotent Stem Cells10.3390/biomedicines91115892227-9059https://doaj.org/article/d09301e338eb479ca80f3f40bd7f44772021-10-01T00:00:00Zhttps://www.mdpi.com/2227-9059/9/11/1589https://doaj.org/toc/2227-9059Ca<sup>2+</sup> overload is one of the factors leading to Duchenne muscular dystrophy (DMD) pathogenesis. However, the molecular targets of dystrophin deficiency-dependent Ca<sup>2+</sup> overload and the correlation between Ca<sup>2+</sup> overload and contractile DMD phenotypes in in vitro human models remain largely elusive. In this study, we utilized DMD patient-derived induced pluripotent stem cells (iPSCs) to differentiate myotubes using doxycycline-inducible MyoD overexpression, and searched for a target molecule that mediates dystrophin deficiency-dependent Ca<sup>2+</sup> overload using commercially available chemicals and siRNAs. We found that several store-operated Ca<sup>2+</sup> channel (SOC) inhibitors effectively prevented Ca<sup>2+</sup> overload and identified that STIM1–Orai1 is a molecular target of SOCs. These findings were further confirmed by demonstrating that STIM1–Orai1 inhibitors, CM4620, AnCoA4, and GSK797A, prevented Ca<sup>2+</sup> overload in dystrophic myotubes. Finally, we evaluated CM4620, AnCoA4, and GSK7975A activities using a previously reported model recapitulating a muscle fatigue-like decline in contractile performance in DMD. All three chemicals ameliorated the decline in contractile performance, indicating that modulating STIM1–Orai1-mediated Ca<sup>2+</sup> overload is effective in rescuing contractile phenotypes. In conclusion, SOCs are major contributors to dystrophin deficiency-dependent Ca<sup>2+</sup> overload through STIM1–Orai1 as molecular mediators. Modulating STIM1–Orai1 activity was effective in ameliorating the decline in contractile performance in DMD.Tomoya UchimuraHidetoshi SakuraiMDPI AGarticleiPSCskeletal muscleCa<sup>2+</sup> overloadstore-operated Ca<sup>2+</sup> channelSTIM1-Orai1Biology (General)QH301-705.5ENBiomedicines, Vol 9, Iss 1589, p 1589 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	iPSC skeletal muscle Ca<sup>2+</sup> overload store-operated Ca<sup>2+</sup> channel STIM1-Orai1 Biology (General) QH301-705.5
spellingShingle	iPSC skeletal muscle Ca<sup>2+</sup> overload store-operated Ca<sup>2+</sup> channel STIM1-Orai1 Biology (General) QH301-705.5 Tomoya Uchimura Hidetoshi Sakurai Orai1–STIM1 Regulates Increased Ca<sup>2+</sup> Mobilization, Leading to Contractile Duchenne Muscular Dystrophy Phenotypes in Patient-Derived Induced Pluripotent Stem Cells
description	Ca<sup>2+</sup> overload is one of the factors leading to Duchenne muscular dystrophy (DMD) pathogenesis. However, the molecular targets of dystrophin deficiency-dependent Ca<sup>2+</sup> overload and the correlation between Ca<sup>2+</sup> overload and contractile DMD phenotypes in in vitro human models remain largely elusive. In this study, we utilized DMD patient-derived induced pluripotent stem cells (iPSCs) to differentiate myotubes using doxycycline-inducible MyoD overexpression, and searched for a target molecule that mediates dystrophin deficiency-dependent Ca<sup>2+</sup> overload using commercially available chemicals and siRNAs. We found that several store-operated Ca<sup>2+</sup> channel (SOC) inhibitors effectively prevented Ca<sup>2+</sup> overload and identified that STIM1–Orai1 is a molecular target of SOCs. These findings were further confirmed by demonstrating that STIM1–Orai1 inhibitors, CM4620, AnCoA4, and GSK797A, prevented Ca<sup>2+</sup> overload in dystrophic myotubes. Finally, we evaluated CM4620, AnCoA4, and GSK7975A activities using a previously reported model recapitulating a muscle fatigue-like decline in contractile performance in DMD. All three chemicals ameliorated the decline in contractile performance, indicating that modulating STIM1–Orai1-mediated Ca<sup>2+</sup> overload is effective in rescuing contractile phenotypes. In conclusion, SOCs are major contributors to dystrophin deficiency-dependent Ca<sup>2+</sup> overload through STIM1–Orai1 as molecular mediators. Modulating STIM1–Orai1 activity was effective in ameliorating the decline in contractile performance in DMD.
format	article
author	Tomoya Uchimura Hidetoshi Sakurai
author_facet	Tomoya Uchimura Hidetoshi Sakurai
author_sort	Tomoya Uchimura
title	Orai1–STIM1 Regulates Increased Ca<sup>2+</sup> Mobilization, Leading to Contractile Duchenne Muscular Dystrophy Phenotypes in Patient-Derived Induced Pluripotent Stem Cells
title_short	Orai1–STIM1 Regulates Increased Ca<sup>2+</sup> Mobilization, Leading to Contractile Duchenne Muscular Dystrophy Phenotypes in Patient-Derived Induced Pluripotent Stem Cells
title_full	Orai1–STIM1 Regulates Increased Ca<sup>2+</sup> Mobilization, Leading to Contractile Duchenne Muscular Dystrophy Phenotypes in Patient-Derived Induced Pluripotent Stem Cells
title_fullStr	Orai1–STIM1 Regulates Increased Ca<sup>2+</sup> Mobilization, Leading to Contractile Duchenne Muscular Dystrophy Phenotypes in Patient-Derived Induced Pluripotent Stem Cells
title_full_unstemmed	Orai1–STIM1 Regulates Increased Ca<sup>2+</sup> Mobilization, Leading to Contractile Duchenne Muscular Dystrophy Phenotypes in Patient-Derived Induced Pluripotent Stem Cells
title_sort	orai1–stim1 regulates increased ca<sup>2+</sup> mobilization, leading to contractile duchenne muscular dystrophy phenotypes in patient-derived induced pluripotent stem cells
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/d09301e338eb479ca80f3f40bd7f4477
work_keys_str_mv	AT tomoyauchimura orai1stim1regulatesincreasedcasup2supmobilizationleadingtocontractileduchennemusculardystrophyphenotypesinpatientderivedinducedpluripotentstemcells AT hidetoshisakurai orai1stim1regulatesincreasedcasup2supmobilizationleadingtocontractileduchennemusculardystrophyphenotypesinpatientderivedinducedpluripotentstemcells
_version_	1718412925444554752

Orai1–STIM1 Regulates Increased Ca<sup>2+</sup> Mobilization, Leading to Contractile Duchenne Muscular Dystrophy Phenotypes in Patient-Derived Induced Pluripotent Stem Cells

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