Impaired Relaxation in Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Pathogenic TNNI3 Mutation of Pediatric Restrictive Cardiomyopathy

Wang, Renjie; Hasegawa, Moyu; Suginobe, Hidehiro; Yoshihara, Chika; Ishii, Yoichiro; Ueyama, Atsuko; Ueda, Kazutoshi; Hashimoto, Kazuhisa; Hirose, Masaki; Ishii, Ryo; Narita, Jun; Watanabe, Takuji; Kawamura, Takuji; Taira, Masaki; Ueno, Takayoshi; Miyagawa, Shigeru; Ishida, Hidekazu

doi:https://doi.org/10.1161/JAHA.123.032375

Impaired Relaxation in Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Pathogenic TNNI3 Mutation of Pediatric Restrictive Cardiomyopathy

Wang, Renjie; Hasegawa, Moyu; Suginobe, Hidehiro et al.

Journal of the American Heart Association, 2024, 13(6), e032375

Number of Access:2,058（2025-08-15 22:51 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/95638

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Title	Impaired Relaxation in Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Pathogenic TNNI3 Mutation of Pediatric Restrictive Cardiomyopathy
Creator	CreatorName Wang, Renjie
	NRID 1000000896508 1000000896508 CreatorName Hasegawa, Moyu
	CreatorName Suginobe, Hidehiro
	CreatorName Yoshihara, Chika
	NRID 1000090573882 1000090573882 CreatorName Ishii, Yoichiro
	CreatorName Ueyama, Atsuko
	CreatorName Ueda, Kazutoshi
	CreatorName Hashimoto, Kazuhisa
	researchmap permalink masahirose masahirose Scopus Author ID 57908743200 57908743200 NRID 1000010973662 1000010973662 CreatorName Hirose, Masaki
	NRID 1000090794008 1000090794008 researchmap permalink iryo-0918 iryo-0918 Scopus Author ID 56017974100 56017974100 CreatorName Ishii, Ryo
	NRID 1000070467562 1000070467562 researchmap permalink J-narita J-narita Scopus Author ID 36993204400 36993204400 CreatorName Narita, Jun
	NRID 1000080838582 1000080838582 researchmap permalink takuji-watanabe takuji-watanabe Scopus Author ID 57209196810 57209196810 CreatorName Watanabe, Takuji
	NRID 1000060839398 1000060839398 researchmap permalink takuji-kawamura takuji-kawamura Scopus Author ID 55331768500 55331768500 ORCID 0000-0003-2985-0036 0000-0003-2985-0036 CreatorName Kawamura, Takuji
	NRID 1000090624651 1000090624651 Scopus Author ID 57188551084 57188551084 CreatorName Taira, Masaki
	NRID 1000060437316 1000060437316 researchmap permalink takayoshi-ueno takayoshi-ueno Scopus Author ID 8137931400 8137931400 CreatorName Ueno, Takayoshi
	NRID 1000070544237 1000070544237 ORCID 0000-0003-0015-6569 0000-0003-0015-6569 researchmap permalink s-miyagawa3154 s-miyagawa3154 Scopus Author ID 55560624700 55560624700 CreatorName Miyagawa, Shigeru
	NRID 1000050467552 1000050467552 researchmap permalink researcher_cardio researcher_cardio Scopus Author ID 36863872000 36863872000 CreatorName Ishida, Hidekazu
Subject	cardiomyocytes
	diastolic dysfunction
	iPSC
	isogenic
	motion analysis
	restrictive cardiomyopathy
Description	Wang R., Hasegawa M., Suginobe H., et al. Impaired Relaxation in Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Pathogenic TNNI3 Mutation of Pediatric Restrictive Cardiomyopathy. Journal of the American Heart Association 13, e032375 (2024); https://doi.org/10.1161/JAHA.123.032375.
Abstract	BACKGROUND: Restrictive cardiomyopathy (RCM) is characterized by impaired diastolic function with preserved ventricular contraction. Several pathogenic variants in sarcomere genes, including TNNI3, are reported to cause Ca²⁺ hypersensitivity in cardiomyocytes in overexpression models; however, the pathophysiology of induced pluripotent stem cell (iPSC)-derived cardiomyocytes specific to a patient with RCM remains unknown. METHODS AND RESULTS: We established an iPSC line from a pediatric patient with RCM and a heterozygous TNNI3 missense variant, c.508C>T (p.Arg170Trp; R170W). We conducted genome editing via CRISPR/Cas9 technology to establish an isogenic correction line harboring wild type TNNI3 as well as a homozygous TNNI3-R170W. iPSCs were then differentiated to cardio-myocytes to compare their cellular physiological, structural, and transcriptomic features. Cardiomyocytes differentiated from heterozygous and homozygous TNNI3-R170W iPSC lines demonstrated impaired diastolic function in cell motion analyses as compared with that in cardiomyocytes derived from isogenic-corrected iPSCs and 3 independent healthy iPSC lines. The intracellular Ca²⁺ oscillation and immunocytochemistry of troponin I were not significantly affected in RCM-cardiomyocytes with either heterozygous or homozygous TNNI3-R170W. Electron microscopy showed that the myofibril and mitochondrial structures appeared to be unaffected. RNA sequencing revealed that pathways associated with cardiac muscle development and contraction, extracellular matrix-receptor interaction, and transforming growth factor-β were altered in RCM-iPSC-derived cardiomyocytes. CONCLUSIONS: Patient-specific iPSC-derived cardiomyocytes could effectively represent the diastolic dysfunction of RCM. Myofibril structures including troponin I remained unaffected in the monolayer culture system, although gene expression profiles associated with cardiac muscle functions were altered.
Publisher	American Heart Association Inc.
Source Title	Journal of the American Heart Association
Volume (Issue)	13(6)
Page	e032375
Date of Issued	2024-03-19
Language	English
Handle URL	https://hdl.handle.net/11094/95638
eISSN	20479980
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1161/JAHA.123.032375
Access Rights	open access
CopyRight	This article is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
oaire:version	Version of Record
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCMI Type	Text
DCTERMS.bibliographicCitation	Journal of the American Heart Association. 2024, 13(6), p. e032375
DC.title	Impaired Relaxation in Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Pathogenic TNNI3 Mutation of Pediatric Restrictive Cardiomyopathy
DC.creator	Wang, Renjie
	Hasegawa, Moyu
	Suginobe, Hidehiro
	Yoshihara, Chika
	Ishii, Yoichiro
	Ueyama, Atsuko
	Ueda, Kazutoshi
	Hashimoto, Kazuhisa
	Hirose, Masaki
	Ishii, Ryo
	Narita, Jun
	Watanabe, Takuji
	Kawamura, Takuji
	Taira, Masaki
	Ueno, Takayoshi
	Miyagawa, Shigeru
	Ishida, Hidekazu
DC.publisher	American Heart Association Inc.
DC.language' scheme='DCTERMS.RFC1766	English
DC.type' scheme='DCTERMS.DCMIType	Text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2024-03-19
DC.identifier	https://doi.org/10.1161/JAHA.123.032375
DC.subject	cardiomyocytes
	diastolic dysfunction
	iPSC
	isogenic
	motion analysis
	restrictive cardiomyopathy
DC.description	Wang R., Hasegawa M., Suginobe H., et al. Impaired Relaxation in Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Pathogenic TNNI3 Mutation of Pediatric Restrictive Cardiomyopathy. Journal of the American Heart Association 13, e032375 (2024); https://doi.org/10.1161/JAHA.123.032375.
DCTERMS.abstract	BACKGROUND: Restrictive cardiomyopathy (RCM) is characterized by impaired diastolic function with preserved ventricular contraction. Several pathogenic variants in sarcomere genes, including TNNI3, are reported to cause Ca²⁺ hypersensitivity in cardiomyocytes in overexpression models; however, the pathophysiology of induced pluripotent stem cell (iPSC)-derived cardiomyocytes specific to a patient with RCM remains unknown. METHODS AND RESULTS: We established an iPSC line from a pediatric patient with RCM and a heterozygous TNNI3 missense variant, c.508C>T (p.Arg170Trp; R170W). We conducted genome editing via CRISPR/Cas9 technology to establish an isogenic correction line harboring wild type TNNI3 as well as a homozygous TNNI3-R170W. iPSCs were then differentiated to cardio-myocytes to compare their cellular physiological, structural, and transcriptomic features. Cardiomyocytes differentiated from heterozygous and homozygous TNNI3-R170W iPSC lines demonstrated impaired diastolic function in cell motion analyses as compared with that in cardiomyocytes derived from isogenic-corrected iPSCs and 3 independent healthy iPSC lines. The intracellular Ca²⁺ oscillation and immunocytochemistry of troponin I were not significantly affected in RCM-cardiomyocytes with either heterozygous or homozygous TNNI3-R170W. Electron microscopy showed that the myofibril and mitochondrial structures appeared to be unaffected. RNA sequencing revealed that pathways associated with cardiac muscle development and contraction, extracellular matrix-receptor interaction, and transforming growth factor-β were altered in RCM-iPSC-derived cardiomyocytes. CONCLUSIONS: Patient-specific iPSC-derived cardiomyocytes could effectively represent the diastolic dysfunction of RCM. Myofibril structures including troponin I remained unaffected in the monolayer culture system, although gene expression profiles associated with cardiac muscle functions were altered.
DC.rights	This article is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
DC.format	95638
citation_title	Impaired Relaxation in Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Pathogenic TNNI3 Mutation of Pediatric Restrictive Cardiomyopathy
citation_author	Wang, Renjie
	Hasegawa, Moyu
	Suginobe, Hidehiro
	Yoshihara, Chika
	Ishii, Yoichiro
	Ueyama, Atsuko
	Ueda, Kazutoshi
	Hashimoto, Kazuhisa
	Hirose, Masaki
	Ishii, Ryo
	Narita, Jun
	Watanabe, Takuji
	Kawamura, Takuji
	Taira, Masaki
	Ueno, Takayoshi
	Miyagawa, Shigeru
	Ishida, Hidekazu
citation_publisher	American Heart Association Inc.
citation_language	English
citation_date	2024-03-19
citation_journal_title	Journal of the American Heart Association
citation_volume	13
citation_issue	6
citation_firstpage	e032375
citation_doi	https://doi.org/10.1161/JAHA.123.032375
citation_public_url	https://hdl.handle.net/11094/95638

Impaired Relaxation in Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Pathogenic TNNI3 Mutation of Pediatric Restrictive Cardiomyopathy

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