SARS-CoV-2 spike receptor-binding domain is internalized and promotes protein ISGylation in human induced pluripotent stem cell-derived cardiomyocytes

Okuno, Shota; Higo, Shuichiro; Kondo, Takumi; Shiba, Mikio; Kameda, Satoshi; Inoue, Hiroyuki; Tabata, Tomoka; Ogawa, Shou; Morishita, Yu; Sun, Congcong; Ishino, Saki; Honda, Tomoyuki; Miyagawa, Shigeru; Sakata, Yasushi

doi:https://doi.org/10.1038/s41598-023-48084-7

SARS-CoV-2 spike receptor-binding domain is internalized and promotes protein ISGylation in human induced pluripotent stem cell-derived cardiomyocytes

Okuno, Shota; Higo, Shuichiro; Kondo, Takumi et al.

Scientific Reports, 2023, 13(1), 21397

アクセス数:1,215件（2025-07-22 20:47 集計）

固定URL: https://hdl.handle.net/11094/93523

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タイトル	SARS-CoV-2 spike receptor-binding domain is internalized and promotes protein ISGylation in human induced pluripotent stem cell-derived cardiomyocytes
著者	著者名 Okuno, Shota
	NRID 1000000604034 1000000604034 researchmap permalink s.higo s.higo Scopus Author ID 23018633200 23018633200 著者名 Higo, Shuichiro
	著者名 Kondo, Takumi
	著者名 Shiba, Mikio
	著者名 Kameda, Satoshi
	著者名 Inoue, Hiroyuki
	NRID 1000030973094 1000030973094 researchmap permalink tomoka0122 tomoka0122 著者名 Tabata, Tomoka
	著者名 Ogawa, Shou
	著者名 Morishita, Yu
	著者名 Sun, Congcong
	著者名 Ishino, Saki
	NRID 1000080402676 1000080402676 著者名 Honda, Tomoyuki
	NRID 1000070544237 1000070544237 ORCID 0000-0003-0015-6569 0000-0003-0015-6569 researchmap permalink s-miyagawa3154 s-miyagawa3154 Scopus Author ID 55560624700 55560624700 著者名 Miyagawa, Shigeru
	NRID 1000000397671 1000000397671 ORCID 0000-0002-5618-4721 0000-0002-5618-4721 researchmap permalink C4589 C4589 Scopus Author ID 35395964700 35395964700 著者名 Sakata, Yasushi
内容	Okuno S., Higo S., Kondo T., et al. SARS-CoV-2 spike receptor-binding domain is internalized and promotes protein ISGylation in human induced pluripotent stem cell-derived cardiomyocytes. Scientific Reports 13, 21397 (2023); https://doi.org/10.1038/s41598-023-48084-7.
抄録	Although an increased risk of myocarditis has been observed after vaccination with mRNA encoding severe acute respiratory syndrome coronavirus 2 spike protein, its underlying mechanism has not been elucidated. This study investigated the direct effects of spike receptor-binding domain (S-RBD) on human cardiomyocytes differentiated from induced pluripotent stem cells (iPSC-CMs). Immunostaining experiments using ACE2 wild-type (WT) and knockout (KO) iPSC-CMs treated with purified S-RBD demonstrated that S-RBD was bound to ACE2 and internalized into the subcellular space in the iPSC-CMs, depending on ACE2. Immunostaining combined with live cell imaging using a recombinant S-RBD fused to the superfolder GFP (S-RBD-sfGFP) demonstrated that S-RBD was bound to the cell membrane, co-localized with RAB5A, and then delivered from the endosomes to the lysosomes in iPSC-CMs. Quantitative PCR array analysis followed by single cell RNA sequence analysis clarified that S-RBD-sfGFP treatment significantly upregulated the NF-kβ pathway-related gene (CXCL1) in the differentiated non-cardiomyocytes, while upregulated interferon (IFN)-responsive genes (IFI6, ISG15, and IFITM3) in the matured cardiomyocytes. S-RBD-sfGFP treatment promoted protein ISGylation, an ISG15-mediated post-translational modification in ACE2-WT-iPSC-CMs, which was suppressed in ACE2-KO-iPSC-CMs. Our experimental study demonstrates that S-RBD is internalized through the endolysosomal pathway, which upregulates IFN-responsive genes and promotes ISGylation in the iPSC-CMs.
出版者	Nature Research
収録物名	Scientific Reports
巻(号)	13(1)
ページ	21397
刊行年月	2023-12-04
言語	英語
ハンドルURL	https://hdl.handle.net/11094/93523
eISSN	20452322
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1038/s41598-023-48084-7
アクセス権	open access
権利情報	This article is licensed under a Creative Commons Attribution 4.0 International License.
出版タイプ	Version of Record
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCMI資源タイプ	text
DCTERMS.bibliographicCitation	Scientific Reports. 2023, 13(1), p. 21397
DC.title	SARS-CoV-2 spike receptor-binding domain is internalized and promotes protein ISGylation in human induced pluripotent stem cell-derived cardiomyocytes
DC.creator	Okuno, Shota
	Higo, Shuichiro
	Kondo, Takumi
	Shiba, Mikio
	Kameda, Satoshi
	Inoue, Hiroyuki
	Tabata, Tomoka
	Ogawa, Shou
	Morishita, Yu
	Sun, Congcong
	Ishino, Saki
	Honda, Tomoyuki
	Miyagawa, Shigeru
	Sakata, Yasushi
DC.publisher	Nature Research
DC.language' scheme='DCTERMS.RFC1766	英語
DC.type' scheme='DCTERMS.DCMIType	text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2023-12-04
DC.identifier	https://doi.org/10.1038/s41598-023-48084-7
DC.description	Okuno S., Higo S., Kondo T., et al. SARS-CoV-2 spike receptor-binding domain is internalized and promotes protein ISGylation in human induced pluripotent stem cell-derived cardiomyocytes. Scientific Reports 13, 21397 (2023); https://doi.org/10.1038/s41598-023-48084-7.
DCTERMS.abstract	Although an increased risk of myocarditis has been observed after vaccination with mRNA encoding severe acute respiratory syndrome coronavirus 2 spike protein, its underlying mechanism has not been elucidated. This study investigated the direct effects of spike receptor-binding domain (S-RBD) on human cardiomyocytes differentiated from induced pluripotent stem cells (iPSC-CMs). Immunostaining experiments using ACE2 wild-type (WT) and knockout (KO) iPSC-CMs treated with purified S-RBD demonstrated that S-RBD was bound to ACE2 and internalized into the subcellular space in the iPSC-CMs, depending on ACE2. Immunostaining combined with live cell imaging using a recombinant S-RBD fused to the superfolder GFP (S-RBD-sfGFP) demonstrated that S-RBD was bound to the cell membrane, co-localized with RAB5A, and then delivered from the endosomes to the lysosomes in iPSC-CMs. Quantitative PCR array analysis followed by single cell RNA sequence analysis clarified that S-RBD-sfGFP treatment significantly upregulated the NF-kβ pathway-related gene (CXCL1) in the differentiated non-cardiomyocytes, while upregulated interferon (IFN)-responsive genes (IFI6, ISG15, and IFITM3) in the matured cardiomyocytes. S-RBD-sfGFP treatment promoted protein ISGylation, an ISG15-mediated post-translational modification in ACE2-WT-iPSC-CMs, which was suppressed in ACE2-KO-iPSC-CMs. Our experimental study demonstrates that S-RBD is internalized through the endolysosomal pathway, which upregulates IFN-responsive genes and promotes ISGylation in the iPSC-CMs.
DC.rights	This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	93523
citation_title	SARS-CoV-2 spike receptor-binding domain is internalized and promotes protein ISGylation in human induced pluripotent stem cell-derived cardiomyocytes
citation_author	Okuno, Shota
	Higo, Shuichiro
	Kondo, Takumi
	Shiba, Mikio
	Kameda, Satoshi
	Inoue, Hiroyuki
	Tabata, Tomoka
	Ogawa, Shou
	Morishita, Yu
	Sun, Congcong
	Ishino, Saki
	Honda, Tomoyuki
	Miyagawa, Shigeru
	Sakata, Yasushi
citation_publisher	Nature Research
citation_language	英語
citation_date	2023-12-04
citation_journal_title	Scientific Reports
citation_volume	13
citation_issue	1
citation_firstpage	21397
citation_doi	https://doi.org/10.1038/s41598-023-48084-7
citation_public_url	https://hdl.handle.net/11094/93523

SARS-CoV-2 spike receptor-binding domain is internalized and promotes protein ISGylation in human induced pluripotent stem cell-derived cardiomyocytes

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