A novel transient receptor potential C3/C6 selective activator induces the cellular uptake of antisense oligonucleotides

Kohashi, Hiroto; Nagata, Ryu; Tamenori, Yusuke; Amatani, Tomorrow; Ueda, Yoshifumi; Mori, Yasuo; Kasahara, Yuuya; Obika, Satoshi; Shimojo, Masahito

doi:https://doi.org/10.1093/nar/gkae245

A novel transient receptor potential C3/C6 selective activator induces the cellular uptake of antisense oligonucleotides

Kohashi, Hiroto; Nagata, Ryu; Tamenori, Yusuke et al.

Nucleic Acids Research, 2024, 52(9), 4784-4798

Number of Access:826（2025-11-27 00:29 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/97215

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Title	A novel transient receptor potential C3/C6 selective activator induces the cellular uptake of antisense oligonucleotides
Creator	CreatorName Kohashi, Hiroto
	CreatorName Nagata, Ryu
	CreatorName Tamenori, Yusuke
	CreatorName Amatani, Tomorrow
	CreatorName Ueda, Yoshifumi
	NRID 1000080212265 1000080212265 CreatorName Mori, Yasuo
	NRID 1000010740673 1000010740673 researchmap permalink YKasa YKasa CreatorName Kasahara, Yuuya
	NRID 1000080243252 1000080243252 ORCID 0000-0002-6842-6812 0000-0002-6842-6812 researchmap permalink read0014368 read0014368 Scopus Author ID 7003800550 7003800550 CreatorName Obika, Satoshi
	NRID 1000090591925 1000090591925 researchmap permalink shimojom shimojom Scopus Author ID 7102952254 7102952254 CreatorName Shimojo, Masahito
Description	Kohashi H., Nagata R., Tamenori Y., et al. A novel transient receptor potential C3/C6 selective activator induces the cellular uptake of antisense oligonucleotides. Nucleic Acids Research 52, 4784 (2024); https://doi.org/10.1093/nar/gkae245.
Abstract	Antisense oligonucleotide (ASO) therapy is a novel therapeutic approach in which ASO specifically binds target mRNA, resulting in mRNA degradation; however, cellular uptake of ASOs remains critically low, warranting improvement. Transient receptor potential canonical (TRPC) channels regulate Ca2+ influx and are activated upon stimulation by phospholipase C-generated diacylglycerol. Herein, we report that a novel TRPC3/C6/C7 activator, L687, can induce cellular ASO uptake. L687-induced ASO uptake was enhanced in a dose- and incubation-time-dependent manner. L687 enhanced the knockdown activity of various ASOs both in vitro and in vivo. Notably, suppression of TRPC3/C6 by specific siRNAs reduced ASO uptake in A549 cells. Application of BAPTA-AM, a Ca2+ chelator, and SKF96365, a TRPC3/C6 inhibitor, suppressed Ca2+ influx via TRPC3/C6, resulting in reduced ASO uptake, thereby suggesting that Ca2+ influx via TRPC3/C6 is critical for L687-mediated increased ASO uptake. L687 also induced dextran uptake, indicating that L687 increased endocytosis. Adding ASO to L687 resulted in endosome accumulation; however, the endosomal membrane disruptor UNC7938 facilitated endosomal escape and enhanced knockdown activity. We discovered a new function for TRPC activators regarding ASO trafficking in target cells. Our findings provide an opportunity to formulate an innovative drug delivery system for the therapeutic development of ASO.
Publisher	Oxford University Press
Source Title	Nucleic Acids Research
Volume (Issue)	52(9)
Page	4784-4798
Date of Issued	2024-05-22
Language	English
Handle URL	https://hdl.handle.net/11094/97215
PISSN	03051048
eISSN	13624962
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1093/nar/gkae245
Access Rights	open access
CopyRight	This article is licensed under a Creative Commons Attribution 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
DCTERMS.bibliographicCitation	Nucleic Acids Research. 2024, 52(9), p. 4784-4798
DC.title	A novel transient receptor potential C3/C6 selective activator induces the cellular uptake of antisense oligonucleotides
DC.creator	Kohashi, Hiroto
	Nagata, Ryu
	Tamenori, Yusuke
	Amatani, Tomorrow
	Ueda, Yoshifumi
	Mori, Yasuo
	Kasahara, Yuuya
	Obika, Satoshi
	Shimojo, Masahito
DC.publisher	Oxford University Press
DC.language' scheme='DCTERMS.RFC1766	English
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2024-05-22
DC.identifier	https://doi.org/10.1093/nar/gkae245
DC.description	Kohashi H., Nagata R., Tamenori Y., et al. A novel transient receptor potential C3/C6 selective activator induces the cellular uptake of antisense oligonucleotides. Nucleic Acids Research 52, 4784 (2024); https://doi.org/10.1093/nar/gkae245.
DCTERMS.abstract	Antisense oligonucleotide (ASO) therapy is a novel therapeutic approach in which ASO specifically binds target mRNA, resulting in mRNA degradation; however, cellular uptake of ASOs remains critically low, warranting improvement. Transient receptor potential canonical (TRPC) channels regulate Ca2+ influx and are activated upon stimulation by phospholipase C-generated diacylglycerol. Herein, we report that a novel TRPC3/C6/C7 activator, L687, can induce cellular ASO uptake. L687-induced ASO uptake was enhanced in a dose- and incubation-time-dependent manner. L687 enhanced the knockdown activity of various ASOs both in vitro and in vivo. Notably, suppression of TRPC3/C6 by specific siRNAs reduced ASO uptake in A549 cells. Application of BAPTA-AM, a Ca2+ chelator, and SKF96365, a TRPC3/C6 inhibitor, suppressed Ca2+ influx via TRPC3/C6, resulting in reduced ASO uptake, thereby suggesting that Ca2+ influx via TRPC3/C6 is critical for L687-mediated increased ASO uptake. L687 also induced dextran uptake, indicating that L687 increased endocytosis. Adding ASO to L687 resulted in endosome accumulation; however, the endosomal membrane disruptor UNC7938 facilitated endosomal escape and enhanced knockdown activity. We discovered a new function for TRPC activators regarding ASO trafficking in target cells. Our findings provide an opportunity to formulate an innovative drug delivery system for the therapeutic development of ASO.
DC.rights	This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	97215
citation_title	A novel transient receptor potential C3/C6 selective activator induces the cellular uptake of antisense oligonucleotides
citation_author	Kohashi, Hiroto
	Nagata, Ryu
	Tamenori, Yusuke
	Amatani, Tomorrow
	Ueda, Yoshifumi
	Mori, Yasuo
	Kasahara, Yuuya
	Obika, Satoshi
	Shimojo, Masahito
citation_publisher	Oxford University Press
citation_language	English
citation_date	2024-05-22
citation_journal_title	Nucleic Acids Research
citation_volume	52
citation_issue	9
citation_firstpage	4784
citation_lastpage	4798
citation_doi	https://doi.org/10.1093/nar/gkae245
citation_public_url	https://hdl.handle.net/11094/97215

A novel transient receptor potential C3/C6 selective activator induces the cellular uptake of antisense oligonucleotides

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