An in vitro model of polycystic liver disease using genome-edited human inducible pluripotent stem cells

Akihide Kamiya; Hiromi Chikada; Kinuyo Ida; Emi Ando; Kota Tsuruya; Tatehiro Kagawa; Yutaka Inagaki

doi:10.1016/j.scr.2018.08.018

An in vitro model of polycystic liver disease using genome-edited human inducible pluripotent stem cells

Stem Cell Res. 2018 Oct:32:17-24. doi: 10.1016/j.scr.2018.08.018. Epub 2018 Aug 24.

Authors

Akihide Kamiya¹, Hiromi Chikada², Kinuyo Ida³, Emi Ando³, Kota Tsuruya⁴, Tatehiro Kagawa⁵, Yutaka Inagaki⁶

Affiliations

¹ Department of Molecular Life Sciences, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa, Japan; Center for Matrix Biology and Medicine, Graduate School of Medicine, Tokai University, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan. Electronic address: kamiyaa@tokai-u.jp.
² Department of Molecular Life Sciences, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa, Japan; Center for Matrix Biology and Medicine, Graduate School of Medicine, Tokai University, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan.
³ Department of Molecular Life Sciences, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa, Japan.
⁴ Department of Molecular Life Sciences, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa, Japan; Division of Gastroenterology, Department of Internal Medicine, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan.
⁵ Division of Gastroenterology, Department of Internal Medicine, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan.
⁶ Center for Matrix Biology and Medicine, Graduate School of Medicine, Tokai University, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan; Department of Regenerative Medicine, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan.

PMID: 30172093
DOI: 10.1016/j.scr.2018.08.018

Abstract

In the developing liver, bile duct structure is formed through differentiation of hepatic progenitor cells (HPC) into cholangiocytes. A subtype of polycystic liver diseases characterized by uncontrolled expansion of bile ductal cells is caused by genetic abnormalities such as in that of protein kinase C substrate 80 K-H (PRKCSH). In this study, we aimed to mimic the disease process in vitro by genome editing of the PRKCSH locus in human inducible pluripotent stem (iPS) cells. A proportion of cultured human iPS cell-derived CD13⁺CD133⁺ HPC differentiated into CD13^- cells. During the subsequent gel embedding culture, CD13^- cells formed bile ductal marker-positive cystic structures with the polarity of epithelial cells. A deletion of PRKCSH gene increased expression of cholangiocytic transcription factors in CD13^- cells and the number of cholangiocytic cyst structure. These results suggest that PRKCSH deficiency promotes the differentiation of HPC-derived cholangiocytes, providing a good in vitro model to analyze the molecular mechanisms underlying polycystic diseases.

Keywords: Cholangiocyte; Genome editing; Human iPS cells; Polycystic liver disease.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

AC133 Antigen / metabolism
CD13 Antigens / metabolism
Calcium-Binding Proteins
Cell Differentiation
Cell Line
Cells, Cultured
Cysts / metabolism*
Cysts / pathology*
Flow Cytometry
Gene Editing / methods*
Glucosidases / deficiency
Glucosidases / genetics
Humans
Intracellular Signaling Peptides and Proteins / deficiency
Intracellular Signaling Peptides and Proteins / genetics
Liver / cytology
Liver Diseases / metabolism*
Liver Diseases / pathology*

Substances

AC133 Antigen
Calcium-Binding Proteins
Intracellular Signaling Peptides and Proteins
Glucosidases
PRKCSH protein, human
CD13 Antigens

Supplementary concepts

Polycystic liver disease