Induced Endothelial Cell-Integrated Liver Assembloids Promote Hepatic Maturation and Therapeutic Effect on Cholestatic Liver Fibrosis

Donggyu Nam; Myung Rae Park; Hyunah Lee; Sung Chul Bae; Daniela Gerovska; Marcos J Araúzo-Bravo; Holm Zaehres; Hans R Schöler; Jeong Beom Kim

doi:10.3390/cells11142242

Induced Endothelial Cell-Integrated Liver Assembloids Promote Hepatic Maturation and Therapeutic Effect on Cholestatic Liver Fibrosis

Cells. 2022 Jul 19;11(14):2242. doi: 10.3390/cells11142242.

Authors

Donggyu Nam¹, Myung Rae Park², Hyunah Lee¹, Sung Chul Bae³, Daniela Gerovska⁴, Marcos J Araúzo-Bravo^{4

5

6

7}, Holm Zaehres^{7

8}, Hans R Schöler⁷, Jeong Beom Kim²

Affiliations

¹ Hans Schöler Stem Cell Research Center (HSSCRC), Department of Biological Sciences, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Korea.
² Hans Schöler Stem Cell Research Center (HSSCRC), Department of Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Korea.
³ Department of Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Korea.
⁴ Group of Computational Biology and Systems Biomedicine, Biodonostia Health Research Institute, 20014 San Sebastian, Spain.
⁵ IKERBASQUE, Basque Foundation for Science, 48013 Bilbao, Spain.
⁶ Department of Cell Biology and Histology, Faculty of Medicine and Nursing, University of Basque Country (UPV/EHU), 48940 Leioa, Spain.
⁷ Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, 48149 Münster, Germany.
⁸ Department of Anatomy and Molecular Embryology, Medical Faculty, Ruhr-University Bochum, 44801 Bochum, Germany.

Abstract

The transplantation of pluripotent stem cell (PSC)-derived liver organoids has been studied to solve the current donor shortage. However, the differentiation of unintended cell populations, difficulty in generating multi-lineage organoids, and tumorigenicity of PSC-derived organoids are challenges. However, direct conversion technology has allowed for the generation lineage-restricted induced stem cells from somatic cells bypassing the pluripotent state, thereby eliminating tumorigenic risks. Here, liver assembloids (iHEAs) were generated by integrating induced endothelial cells (iECs) into the liver organoids (iHLOs) generated with induced hepatic stem cells (iHepSCs). Liver assembloids showed enhanced functional maturity compared to iHLOs in vitro and improved therapeutic effects on cholestatic liver fibrosis animals in vivo. Mechanistically, FN1 expressed from iECs led to the upregulation of Itgα5/β1 and Hnf4α in iHEAs and were correlated to the decreased expression of genes related to hepatic stellate cell activation such as Lox and Spp1 in the cholestatic liver fibrosis animals. In conclusion, our study demonstrates the possibility of generating transplantable iHEAs with directly converted cells, and our results evidence that integrating iECs allows iHEAs to have enhanced hepatic maturation compared to iHLOs.

Keywords: assembloid; assembloid transplantation; cholestatic liver fibrosis; direct conversion; induced endothelial cells; induced hepatic stem cells; organoid.

Publication types

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

MeSH terms

Animals
Cholestasis* / metabolism
Endothelial Cells*
Liver Cirrhosis / metabolism
Organoids / metabolism

Grants and funding

This study was supported by the Leading Foreign Research Institute Recruitment Program (2018K1A4A3A01063890) through the National Research Foundation of Korea (NRF), and 2022 INNOPOLIS Technology Transfer (R&BD) Program (2022-IT-RD-0139) of the INNOPOLIS Korea Innovation foundation funded by the Ministry of Science and ICT, Smart healthcare research center at UNIST (Ulsan National Institute of Science & Technology), and SuPine Therapeutics Inc. M.J.A.-B. was supported by grants from the Ministry of Economy and Competitiveness, Spain; MINECO Grant No. PID2020-119715GB-I00 co-funded by the European Regional Development Fund (ERDF/ESF, Investing in your future).