Efficacy of primary liver organoid culture from different stages of non-alcoholic steatohepatitis (NASH) mouse model

Mohamed Elbadawy; Megumi Yamanaka; Yuta Goto; Kimika Hayashi; Ryouichi Tsunedomi; Shoichi Hazama; Hiroaki Nagano; Toshinori Yoshida; Makoto Shibutani; Ryo Ichikawa; Junta Nakahara; Tsutomu Omatsu; Tetsuya Mizutani; Yukie Katayama; Yuta Shinohara; Amira Abugomaa; Masahiro Kaneda; Hideyuki Yamawaki; Tatsuya Usui; Kazuaki Sasaki

doi:10.1016/j.biomaterials.2020.119823

Efficacy of primary liver organoid culture from different stages of non-alcoholic steatohepatitis (NASH) mouse model

Biomaterials. 2020 Apr:237:119823. doi: 10.1016/j.biomaterials.2020.119823. Epub 2020 Jan 27.

Authors

Mohamed Elbadawy¹, Megumi Yamanaka², Yuta Goto², Kimika Hayashi², Ryouichi Tsunedomi³, Shoichi Hazama⁴, Hiroaki Nagano³, Toshinori Yoshida⁵, Makoto Shibutani⁵, Ryo Ichikawa⁵, Junta Nakahara⁵, Tsutomu Omatsu⁶, Tetsuya Mizutani⁶, Yukie Katayama⁶, Yuta Shinohara⁷, Amira Abugomaa², Masahiro Kaneda⁸, Hideyuki Yamawaki⁹, Tatsuya Usui¹⁰, Kazuaki Sasaki²

Affiliations

¹ Laboratory of Veterinary Pharmacology, Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan; Department of Pharmacology, Faculty of Veterinary Medicine, Benha University, 13736, Moshtohor, Toukh, Elqaliobiya, Egypt.
² Laboratory of Veterinary Pharmacology, Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan.
³ Department of Gastroenterological, Breast and Endocrine Surgery, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi, 755-8505, Japan.
⁴ Department of Gastroenterological, Breast and Endocrine Surgery, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi, 755-8505, Japan; Department of Translational Research and Developmental Therapeutics Against Cancer, Yamaguchi University School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi, 755-8505, Japan.
⁵ Laboratory of Veterinary Pathology, Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan.
⁶ Research and Education Center for Prevention of Global Infectious Disease of Animals, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan.
⁷ Laboratory of Veterinary Pharmacology, Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan; Pet Health & Food Division, Iskara Industry CO., LTD, 1-14-2, Nihonbashi, Chuo-ku, Tokyo, 103-0027, Japan.
⁸ Laboratory of Veterinary Anatomy, Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan.
⁹ Laboratory of Veterinary Pharmacology, School of Veterinary Medicine, Kitasato University, 35-1, Higashi 23 Ban-cho, Towada, Aomori, 034-8628, Japan.
¹⁰ Laboratory of Veterinary Pharmacology, Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan. Electronic address: fu7085@go.tuat.ac.jp.

PMID: 32044522
DOI: 10.1016/j.biomaterials.2020.119823

Abstract

Non-alcoholic steatohepatitis (NASH) is associated with liver fibrosis and cirrhosis, which eventually leads to hepatocellular carcinoma. Although several animal models were developed to understand the mechanisms of NASH pathogenesis and progression, it remains obscure. A 3D organoid culture system can recapitulate organ structures and maintain gene expression profiles of original tissues. We therefore tried to generate liver organoids from different degrees [defined as mild (NASH A), moderate (NASH B) and severe (NASH C)] of methionine- and choline-deficient diet-induced NASH model mice and analyzed the difference of their architecture, cell components, organoid-forming efficacy, and gene expression profiles. Organoids from each stage of NASH model mice were successfully generated. Interestingly, epithelial-mesenchymal transition was observed in NASH C organoids. Expression of Collagen I and an activated hepatic stellite cell marker, α-sma was upregulated in the liver organoids from NASH B and C mice. The analysis of RNA sequencing revealed that several novel genes were upregulated in all NASH liver organoids. These results suggest that our generated liver organoids from different stages of NASH diseased mice might become a useful tool for in vitro studies of the molecular mechanism of NASH development and also for identifying novel biomarkers for early diagnosis of NASH disease.

Keywords: Biomarker; Liver organoid; NASH; RNA seq, EMT, Liver fibrosis.

Publication types

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

MeSH terms

Animals
Disease Models, Animal
Liver / pathology
Liver Cirrhosis / pathology
Liver Neoplasms* / pathology
Mice
Mice, Inbred C57BL
Non-alcoholic Fatty Liver Disease*
Organoids