EXPRESSION PROFILES OF DRUG-METABOLIZING ENZYME CYP3A AND DRUG EFFLUX TRANSPORTER MULTIDRUG RESISTANCE 1 SUBFAMILY mRNAS IN RAT SMALL INTESTINE

The endoderm, differentiated from human induced pluripotent stem cells (iPSCs), can differentiate into the small intestine and liver, which are vital for drug absorption and metabolism. The development of human iPSC-derived enterocytes (HiEnts) and hepatocytes (HiHeps) has been reported. However, pharmacokinetic function-deficiency of these cells remains to be elucidated. Here, we aimed to develop an efficient differentiation method to induce endoderm formation from human iPSCs. Cells treated with activin A for 168 h expressed higher levels of endodermal genes than those treated for 72 h. Using activin A (days 0–7), CHIR99021 and PI−103 (days 0–2), and FGF2 (days 3–7), the hiPSC-derived endoderm (HiEnd) showed 97.97% CD−117 and CD−184 double-positive cells. Moreover, HiEnts derived from the human iPSC line Windy had similar or higher expression of small intestine-specific genes than adult human small intestine. Activities of the drug transporter P-glycoprotein and drug-metabolizing enzyme cytochrome P450 (CYP) 3A4/5 were confirmed. Additionally, Windy-derived HiHeps expressed higher levels of hepatocyte- and pharmacokinetics-related genes and proteins and showed higher CYP3A4/5 activity than those derived through the conventional differentiation method. Thus, using this novel method, the differentiated HiEnts and HiHeps with pharmacokinetic functions could be used for drug development.

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Generation of tetracycline-controllable CYP3A4-expressing Caco-2 cells by the piggyBac transposon system

Scientific Reports ◽

10.1038/s41598-021-91160-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Moe Ichikawa ◽

Hiroki Akamine ◽

Michika Murata ◽

Sumito Ito ◽

Kazuo Takayama ◽

...

Keyword(s):

Small Intestine ◽

Drug Absorption ◽

Cell Model ◽

Negative Effects ◽

Piggybac Transposon ◽

Drug Metabolizing Enzyme ◽

Metabolizing Enzyme ◽

Cyp3a4 Expression

AbstractCaco-2 cells are widely used as an in vitro intestinal epithelial cell model because they can form a monolayer and predict drug absorption with high accuracy. However, Caco-2 cells hardly express cytochrome P450 (CYP), a drug-metabolizing enzyme. It is known that CYP3A4 is the dominant drug-metabolizing enzyme in human small intestine. In this study, we generated CYP3A4-expressing Caco-2 (CYP3A4-Caco-2) cells and attempted to establish a model that can simultaneously evaluate drug absorption and metabolism. CYP3A4-Caco-2 cells were generated by piggyBac transposon vectors. A tetracycline-controllable CYP3A4 expression cassette (tet-on system) was stably transduced into Caco-2 cells, thus regulating the levels of CYP3A4 expression depending on the doxycycline concentration. The CYP3A4 expression levels in CYP3A4-Caco-2 cells cultured in the presence of doxycycline were similar to or higher than those of adult small intestine. The CYP3A4-Caco-2 cells had enough ability to metabolize midazolam, a substrate of CYP3A4. CYP3A4 overexpression had no negative effects on cell proliferation, barrier function, and P-glycoprotein activity in Caco-2 cells. Thus, we succeeded in establishing Caco-2 cells with CYP3A4 metabolizing activity comparable to in vivo human intestinal tissue. This cell line would be useful in pharmaceutical studies as a model that can simultaneously evaluate drug absorption and metabolism.

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Brivanib Exhibits Potential for Pharmacokinetic Drug–Drug Interactions and the Modulation of Multidrug Resistance through the Inhibition of Human ABCG2 Drug Efflux Transporter and CYP450 Biotransformation Enzymes

Molecular Pharmaceutics ◽

10.1021/acs.molpharmaceut.9b00361 ◽

2019 ◽

Vol 16 (11) ◽

pp. 4436-4450 ◽

Cited By ~ 2

Author(s):

Jakub Hofman ◽

Ales Sorf ◽

Dimitrios Vagiannis ◽

Simona Sucha ◽

Sarah Kammerer ◽

...

Keyword(s):

Multidrug Resistance ◽

Drug Interactions ◽

Drug Efflux ◽

Efflux Transporter ◽

Biotransformation Enzymes ◽

Pharmacokinetic Drug

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Orange Juice and Its Component, Hesperidin, Decrease the Expression of Multidrug Resistance-Associated Protein 2 in Rat Small Intestine and Liver

Journal of Biomedicine and Biotechnology ◽

10.1155/2011/502057 ◽

2011 ◽

Vol 2011 ◽

pp. 1-8 ◽

Cited By ~ 8

Author(s):

Minoru Watanabe ◽

Naoki Matsumoto ◽

Yuko Takeba ◽

Toshio Kumai ◽

Masami Tanaka ◽

...

Keyword(s):

Small Intestine ◽

Multidrug Resistance ◽

Orange Juice ◽

Tap Water ◽

Pharmacokinetic Parameters ◽

Mrna Levels ◽

Rat Small Intestine ◽

Sprague Dawley ◽

Water Ingestion ◽

Sprague Dawley Rats

We investigated the effects of orange juice (OJ) or hesperidin, a component of OJ, on the pharmacokinetics of pravastatin (PRV) and the expression of both protein and mRNA of multidrug resistance-associated protein 2 (Mrp2) in the rat small intestine and liver. Eight-week-old male Sprague-Dawley rats were used in this study. OJ or a 0.079% hesperidin suspension was administered orally for 2 days. Tap water was given as a control. A single dose of PRV at 100 mg/kg p.o. was administered after 2 days of OJ, hesperidin, or tap water ingestion. The AUC, , andt1/2values of PRV were significantly increased in OJ group. Mrp2 protein and mRNA levels in the small intestine and liver, respectively, were significantly decreased after the ingestion of OJ. The same results were obtained with hesperidin. These results suggest that the changes in PRV pharmacokinetic parameters and the decrease in Mrp2 expression caused by OJ are due to hesperidin in the juice.

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Integrating miRNA and mRNA expression profiles in response to heat stress-induced injury in rat small intestine

Functional & Integrative Genomics ◽

10.1007/s10142-010-0198-8 ◽

2010 ◽

Vol 11 (2) ◽

pp. 203-213 ◽

Cited By ~ 46

Author(s):

Jin Yu ◽

Fenghua Liu ◽

Peng Yin ◽

Xiaoyu Zhu ◽

Guiling Cheng ◽

...

Keyword(s):

Small Intestine ◽

Heat Stress ◽

Mrna Expression ◽

Expression Profiles ◽

Rat Small Intestine

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Entrectinib reverses cytostatic resistance through the inhibition of ABCB1 efflux transporter, but not the CYP3A4 drug-metabolizing enzyme

Biochemical Pharmacology ◽

10.1016/j.bcp.2020.114061 ◽

2020 ◽

Vol 178 ◽

pp. 114061

Author(s):

Dimitrios Vagiannis ◽

Zhang Yu ◽

Eva Novotna ◽

Anselm Morell ◽

Jakub Hofman

Keyword(s):

Efflux Transporter ◽

Drug Metabolizing Enzyme ◽

Metabolizing Enzyme

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Microarray Analysis of Gene Expression Profiles of Rat Small Intestine in Response to Heat Stress

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057111403928 ◽

2011 ◽

Vol 16 (6) ◽

pp. 655-667 ◽

Cited By ~ 10

Author(s):

An Lu ◽

Huichuan Wang ◽

Xiaolin Hou ◽

Huanrong Li ◽

Guilin Cheng ◽

...

Keyword(s):

Gene Expression ◽

Heat Treatment ◽

Small Intestine ◽

Heat Stress ◽

Differentially Expressed Genes ◽

Microarray Analysis ◽

Expression Profiles ◽

Differentially Expressed ◽

Epithelial Tissue ◽

Rat Small Intestine

Ambient temperature is a critical factor that affects biological organisms in many ways. In this study, the authors investigated gene expression changes in rat small intestine in response to heat stress. Male Sprague-Dawley rats were randomly divided into control and heat-stressed groups. Both groups were housed at 25 °C, although the heat-stressed group was also subjected to 40 °C for 2 h each day for 10 successive days. Rats were sacrificed 1, 3, 6, and 10 days after heat treatment, and sections of their small intestine epithelial tissue were excised for morphological examination and microarray analyses. The rat rectal and body surface temperatures and serum cortisol levels were all significantly increased after heat treatment (p < 0.05). The jejuna were significantly damaged by 3 days after heat treatment began. Microarray analysis showed that 422 genes were differentially expressed, of which 290 genes were significantly upregulated and 132 genes were significantly downregulated. Subsequent bioinformatics analyses revealed that the differentially expressed genes were mainly related to stress, immune regulation, and metabolism processes. The bioinformatics analysis of the differentially expressed genes should be beneficial to further investigations on the underlying mechanisms involved in heat stress–induced damage in the small intestine.

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