title: Inhibition of mitogen-activated protein kinase kinase, DNA methyltransferase, and transforming growth factor-β promotes differentiation of human induced pluripotent stem cells into enterocytes.
abstract: We previously reported that small-molecule compounds were effective in generating pharmacokinetically functional enterocytes from human induced pluripotent stem (iPS) cells. In this study, to determine whether the compounds promote the differentiation of human iPS cells into enterocytes, we investigated the effects of a combination of mitogen-activated protein kinase kinase (MEK), DNA methyltransferase (DNMT), and transforming growth factor (TGF)-β inhibitors on intestinal differentiation. Human iPS cells cultured on feeder cells were differentiated into endodermal cells by activin A. These endodermal-like cells were then differentiated into intestinal stem cells by fibroblast growth factor 2. Finally, the cells were differentiated into enterocyte cells by epidermal growth factor and small-molecule compounds. After differentiation, mRNA expression levels and drug-metabolizing enzyme activities were measured. The mRNA expression levels of the enterocyte marker sucrase-isomaltase and the major drug-metabolizing enzyme cytochrome P450 (CYP) 3A4 were increased by a combination of MEK, DNMT, and TGF-β inhibitors. The mRNA expression of CYP3A4 was markedly induced by 1α,25-dihydroxyvitamin D3. Metabolic activities of CYP1A1/2, CYP2B6, CYP2C9, CYP2C19, CYP3A4/5, UDP-glucuronosyltransferase, and sulfotransferase were also observed in the differentiated cells. In conclusion, MEK, DNMT, and TGF-β inhibitors can be used to promote the differentiation of human iPS cells into pharmacokinetically functional enterocytes.
authors.affiliation: Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, 467-8603, Japan.
authors.forename: Nao
authors.initials: N
authors.lastname: Kodama
authors.affiliation: Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, 467-8603, Japan. Electronic address: tiwao@phar.nagoya-cu.ac.jp.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Iwao
authors.affiliation: Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, 467-8603, Japan.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Kabeya
authors.affiliation: DMPK Research Laboratory, Mitsubishi Tanabe Pharma Corporation, Saitama, 335-8505, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Horikawa
authors.affiliation: DMPK Research Laboratory, Mitsubishi Tanabe Pharma Corporation, Saitama, 335-8505, Japan.
authors.forename: Takuro
authors.initials: T
authors.lastname: Niwa
authors.affiliation: Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, 467-8603, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kondo
authors.affiliation: Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, 467-8603, Japan.
authors.forename: Katsunori
authors.initials: K
authors.lastname: Nakamura
authors.affiliation: Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, 467-8603, Japan.
authors.forename: Tamihide
authors.initials: T
authors.lastname: Matsunaga
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors*; D020895:Enterocytes / Q000201:enzymology / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors*; D012333:RNA, Messenger / Q000096:biosynthesis; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors*
chemical_list: D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D003577:Cytochrome P-450 Enzyme System; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D015254:DNA Modification Methylases; D020929:Mitogen-Activated Protein Kinase Kinases
keywords: DNA methyltransferase inhibitors; Differentiation; Drug-metabolizing enzyme activity; Enterocytes; Human induced pluripotent stem cells; Mitogen-activated protein kinase kinase inhibitors; Small-molecule compounds; Transforming growth factor-β inhibitors