Establishment of SLC15A1/PEPT1-Knockout Human-Induced Pluripotent Stem Cell Line for Intestinal Drug Absorption Studies

  • Mol Ther Methods Clin Dev. 2019 Nov 21;17:49-57. doi: 10.1016/j.omtm.2019.11.008.
Kanae Kawai  1 ,  Ryosuke Negoro  2 ,  Moe Ichikawa  1 ,  Tomoki Yamashita  2 ,  Sayaka Deguchi  2 ,  Kazuo Harada  3 ,  Kazumasa Hirata  3 ,  Kazuo Takayama  1  2  4  5 ,  Hiroyuki Mizuguchi  1  2  5  6  7
Affiliations
  • 1. Laboratory of Biochemistry and Molecular Biology, School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
  • 2. Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
  • 3. Laboratory of Applied Environmental Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
  • 4. PRESTO, Japan Science and Technology Agency, Saitama 332-0012, Japan.
  • 5. Laboratory of Hepatocyte Regulation, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka 567-0085, Japan.
  • 6. Global Center for Medical Engineering and Informatics, Osaka University, Osaka 565-0871, Japan.
  • 7. Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University, Osaka 565-0871, Japan.
PMID: 31890740 DOI: 10.1016/j.omtm.2019.11.008
Abstract

Because many peptide and peptide-mimetic drugs are substrates of peptide transporter 1, it is important to evaluate the peptide transporter 1-mediated intestinal absorption of drug candidates in the early phase of drug development. Although intestinal cell lines treated with inhibitors of peptide transporter 1 are widely used to examine whether drug candidates are substrates for peptide transporter 1, these inhibitors are not sufficiently specific for peptide transporter 1. In this study, to generate a more precise evaluation model, we established peptide transporter 1-knockout induced pluripotent stem cells (iPSCs) by using a CRISPR-Cas9 system and differentiated the cells into intestinal epithelial-like cells. The permeability value and uptake capacity of glycylsarcosine (substrate of peptide transporter 1) in peptide transporter 1-knockout intestinal epithelial-like cells were significantly lower than those in wild-type intestinal epithelial-like cells, suggesting that peptide transporter 1 was successfully depleted in the epithelial cells. Taken together, our model can be useful in the development of peptide and peptide-mimetic drugs.

Keywords
CRISPR-Cas9; PEPT1; SLC15A1; drug absorption; drug transporter; genome editing; human iPS cells; intestinal epithelial-like cells.
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