Generation and application of CES1-knockout Tet-Off-regulated CYP3A4 and UGT1A1-expressing Caco-2 cells

Murata, Michika; Okada, Kentaro; Takahashi, Masaki; Ueyama-Toba, Yukiko; Ito, Sumito; Mizuguchi, Hiroyuki

doi:https://doi.org/10.1016/j.toxlet.2024.10.003

Generation and application of CES1-knockout Tet-Off-regulated CYP3A4 and UGT1A1-expressing Caco-2 cells

Murata, Michika; Okada, Kentaro; Takahashi, Masaki et al.

Toxicology Letters, 2024, 401, 158-169

Number of Access:740（2025-08-16 14:04 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/98590

Link to primary information

File	Format	Terms of use	Size	Views	Date.Available	Description	information
ToxicolLett_401_158	pdf	None	2.00 MB	69	2024.12.13

Creative Commons : Attribution - NonCommercial - NoDerivatives

Item Information

Output File Export EndNote Basic Export Mendeley

Title	Generation and application of CES1-knockout Tet-Off-regulated CYP3A4 and UGT1A1-expressing Caco-2 cells
Creator	CreatorName Murata, Michika
	CreatorName Okada, Kentaro
	CreatorName Takahashi, Masaki
	NRID 1000010904930 1000010904930 ORCID 0000-0002-3491-7955 0000-0002-3491-7955 researchmap permalink toba-y toba-y CreatorName Ueyama-Toba, Yukiko
	NRID 1000020728969 1000020728969 CreatorName Ito, Sumito
	NRID 1000050311387 1000050311387 researchmap permalink read0165079 read0165079 Scopus Author ID 57195381326 57195381326 CreatorName Mizuguchi, Hiroyuki
Subject	Caco-2 cells
	CES
	CYP3A4
	Drug metabolism
	UGT1A1
Description	Murata M., Okada K., Takahashi M., et al. Generation and application of CES1-knockout Tet-Off-regulated CYP3A4 and UGT1A1-expressing Caco-2 cells. Toxicology Letters 401, 158 (2024); https://doi.org/10.1016/j.toxlet.2024.10.003.
Abstract	Caco-2 cells, a human colorectal adenocarcinoma cell line, are widely used to model small intestinal epithelial cells in the drug development process because they can predict drug absorption with high accuracy. However, Caco-2 cells have several issues. First, Caco-2 cells have little expression of cytochrome P450 3A4 (CYP3A4), which is a major drug-metabolizing enzyme in the human intestine. We previously developed Caco-2 cells whose expression of CYP3A4 can be controlled using doxycycline (Dox) (CYP3A4-Caco-2 cells) (Ichikawa et al., Sci. Rep, 2021). However, since the Tet-On system was used to regulate CYP3A4 expression in these cells, there was concern about drug-drug interactions. The second issue is that in the human small intestine, carboxylesterase 2 (CES2) is more highly expressed than carboxylesterase 1 (CES1), while in Caco-2 cells CES1 is more highly expressed. The third issue is the low level expression of uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1), a phase II drug-metabolizing enzyme. In this study, we used genome-editing technology to establish CES1-knockout Caco-2 cells whose CYP3A4 and UGT1A1 expression can be regulated by the Tet-Off system. These cell lines would be useful in pharmaceutical researches, including intestinal toxicological studies, as an in vitro model for orally administered drugs.
Publisher	Elsevier Ireland Ltd
Source Title	Toxicology Letters
Volume (Issue)	401
Page	158-169
Date of Issued	2024-11-01
Language	English
Handle URL	https://hdl.handle.net/11094/98590
PISSN	03784274
eISSN	18793169
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1016/j.toxlet.2024.10.003
Access Rights	open access
CopyRight	This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 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	Toxicology Letters. 2024, 401, p. 158-169
DC.title	Generation and application of CES1-knockout Tet-Off-regulated CYP3A4 and UGT1A1-expressing Caco-2 cells
DC.creator	Murata, Michika
	Okada, Kentaro
	Takahashi, Masaki
	Ueyama-Toba, Yukiko
	Ito, Sumito
	Mizuguchi, Hiroyuki
DC.publisher	Elsevier Ireland Ltd
DC.language' scheme='DCTERMS.RFC1766	English
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2024-11-01
DC.identifier	https://doi.org/10.1016/j.toxlet.2024.10.003
DC.subject	Caco-2 cells
	CES
	CYP3A4
	Drug metabolism
	UGT1A1
DC.description	Murata M., Okada K., Takahashi M., et al. Generation and application of CES1-knockout Tet-Off-regulated CYP3A4 and UGT1A1-expressing Caco-2 cells. Toxicology Letters 401, 158 (2024); https://doi.org/10.1016/j.toxlet.2024.10.003.
DCTERMS.abstract	Caco-2 cells, a human colorectal adenocarcinoma cell line, are widely used to model small intestinal epithelial cells in the drug development process because they can predict drug absorption with high accuracy. However, Caco-2 cells have several issues. First, Caco-2 cells have little expression of cytochrome P450 3A4 (CYP3A4), which is a major drug-metabolizing enzyme in the human intestine. We previously developed Caco-2 cells whose expression of CYP3A4 can be controlled using doxycycline (Dox) (CYP3A4-Caco-2 cells) (Ichikawa et al., Sci. Rep, 2021). However, since the Tet-On system was used to regulate CYP3A4 expression in these cells, there was concern about drug-drug interactions. The second issue is that in the human small intestine, carboxylesterase 2 (CES2) is more highly expressed than carboxylesterase 1 (CES1), while in Caco-2 cells CES1 is more highly expressed. The third issue is the low level expression of uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1), a phase II drug-metabolizing enzyme. In this study, we used genome-editing technology to establish CES1-knockout Caco-2 cells whose CYP3A4 and UGT1A1 expression can be regulated by the Tet-Off system. These cell lines would be useful in pharmaceutical researches, including intestinal toxicological studies, as an in vitro model for orally administered drugs.
DC.rights	This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
DC.format	98590
citation_title	Generation and application of CES1-knockout Tet-Off-regulated CYP3A4 and UGT1A1-expressing Caco-2 cells
citation_author	Murata, Michika
	Okada, Kentaro
	Takahashi, Masaki
	Ueyama-Toba, Yukiko
	Ito, Sumito
	Mizuguchi, Hiroyuki
citation_publisher	Elsevier Ireland Ltd
citation_language	English
citation_date	2024-11-01
citation_journal_title	Toxicology Letters
citation_volume	401
citation_firstpage	158
citation_lastpage	169
citation_doi	https://doi.org/10.1016/j.toxlet.2024.10.003
citation_public_url	https://hdl.handle.net/11094/98590

Generation and application of CES1-knockout Tet-Off-regulated CYP3A4 and UGT1A1-expressing Caco-2 cells

Link to primary information

Item Information

Show details