Generation of Human iPSC-Derived Intestinal Epithelial Cell Monolayers by CDX2 Transduction

Cell Mol Gastroenterol Hepatol. 2019;8(3):513-526. doi: 10.1016/j.jcmgh.2019.06.004. Epub 2019 Jun 19.

Abstract

Background & aims: To develop an effective and safe orally administered drug, it is important to predict its intestinal absorption rate, intestinal first-pass effect, and drug-drug interactions of orally administered drugs. However, there is no existing model to comprehensively predict the intestinal pharmacokinetics and drug-response of orally administered drugs. In this study, we attempted to generate homogenous and functional intestinal epithelial cells from human induced pluripotent stem (iPS) cells for pharmaceutical research.

Methods: We generated almost-homogenous Villin- and zonula occludens-1 (ZO1)-positive intestinal epithelial cells by caudal-related homeobox transcription factor 2 (CDX2) transduction into human iPS cell-derived intestinal progenitor cells.

Results: The drug absorption rates in human iPS cell-derived intestinal epithelial cell monolayers (iPS-IECM) were highly correlated with those in humans (R2=0.91). The expression levels of cytochrome P450 (CYP) 3A4, a dominant drug-metabolizing enzyme in the small intestine, in human iPS-IECM were similar to those in human small intestine in vivo. In addition, intestinal availability in human iPS-IECM (the fraction passing the gut wall: Fg=0.73) was more similar to that in the human small intestine in vivo (Fg=0.57) than to that in Caco-2 cells (Fg=0.99), a human colorectal adenocarcinoma cell line. Moreover, the drug-drug interaction and drug-food interaction could be observed by using our human iPS-IECM in the presence of an inducer and inhibitor of CYP3A4, i.e., rifampicin and grape fruit juice, respectively.

Conclusion: Taking these results together, we succeeded in generating the human iPS-IECM that can be applied to various intestinal pharmacokinetics and drug-response tests of orally administered drugs.

Keywords: Adenovirus; CYP3A4; Differentiation; Intestinal First-Pass Effect.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CDX2 Transcription Factor / genetics*
  • CDX2 Transcription Factor / metabolism
  • Caco-2 Cells
  • Cell Culture Techniques
  • Cell Differentiation
  • Cells, Cultured
  • Cytochrome P-450 CYP3A / metabolism
  • Drug Interactions
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism
  • Food-Drug Interactions
  • Fruit and Vegetable Juices
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Intestinal Absorption
  • Intestines / cytology*
  • Rifampin / pharmacokinetics
  • Transduction, Genetic / methods*

Substances

  • CDX2 Transcription Factor
  • CDX2 protein, human
  • Cytochrome P-450 CYP3A
  • CYP3A4 protein, human
  • Rifampin