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Cited by 378Drug-Induced Hepatotoxicity and ProtectionAnimal testing and alternativesPluripotent Stem Cells Research

High-Fidelity Drug-Induced Liver Injury Screen Using Human Pluripotent Stem Cell–Derived Organoids

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Shinozawa, Tadahiro, Kimura, Masaki, Cai, Yuqi, Saiki, Norikazu, Yoneyama, Yosuke, Ouchi, Rie, Koike, Hiroyuki, Maezawa, Mari +8 more·2021·Gastroenterology
DOI 10.1053/j.gastro.2020.10.002PMC PMC7878295
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Abstract

BACKGROUND & AIMS: Preclinical identification of compounds at risk of causing drug induced liver injury (DILI) remains a significant challenge in drug development, highlighting a need for a predictive human system to study complicated DILI mechanism and susceptibility to individual drug. Here, we established a human liver organoid (HLO)-based screening model for analyzing DILI pathology at organoid resolution. METHODS: We first developed a reproducible method to generate HLO from storable foregut progenitors from pluripotent stem cell (PSC) lines with reproducible bile transport function. The qRT-PCR and single cell RNA-seq determined hepatocyte transcriptomic state in cells of HLO relative to primary hepatocytes. Histological and ultrastructural analyses were performed to evaluate micro-anatomical architecture. HLO based drug-induced liver injury assays were transformed into a 384 well based high-speed live imaging platform. RESULTS: HLO, generated from 10 different pluripotent stem cell lines, contain polarized immature hepatocytes with bile canaliculi-like architecture, establishing the unidirectional bile acid transport pathway. Single cell RNA-seq profiling identified diverse and zonal hepatocytic populations that in part emulate primary adult hepatocytes. The accumulation of fluorescent bile acid into organoid was impaired by CRISPR-Cas9-based gene editing and transporter inhibitor treatment with BSEP. Furthermore, we successfully developed an organoid based assay with multiplexed readouts measuring viability, cholestatic and/or mitochondrial toxicity with high predictive values for 238 marketed drugs at 4 different concentrations (Sensitivity: 88.7%, Specificity: 88.9%). LoT positively predicts genomic predisposition (CYP2C9∗2) for Bosentan-induced cholestasis. CONCLUSIONS: Liver organoid-based Toxicity screen (LoT) is a potential assay system for liver toxicology studies, facilitating compound optimization, mechanistic study, and precision medicine as well as drug screening applications.

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  • iHLOs enable patient-specific modeling for precision medicine tailored to individual genetic and pathological conditions.

    “Furthermore, iHLOs enable patient-specific modeling, providing a platform for precision medicine tailored to individual genetic and pathological conditions12,13.”

    From ​Temporal proteomic profiling of iPSC-derived human liver organoids reveals optimal maturation for drug metabolism and toxicology (2026) · cites this paper as [13]

    The Introduction (page unavailable) says HLOs modeled genotype-specific susceptibility to bosentan-induced cholestasis and could facilitate personalized medicine. The Discussion (page unavailable) describes precision studies using donor-derived HLOs, but frames genomic stratification as a future possibility. The paper does not establish patient-specific modeling tailored to a broad range of individual pathological conditions. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

  • Day 21 or around this timepoint has been identified as an important stage for liver organoid functional development in several previous studies.

    “It is also noteworthy that Day 21, or around this timepoint, has been identified as an important stage for liver organoid functional development in several previous studies13,15,28,29.”

    From ​Temporal proteomic profiling of iPSC-derived human liver organoids reveals optimal maturation for drug metabolism and toxicology (2026) · cites this paper as [13]

    Materials and methods (Large scale screening with 238 test compounds; page unavailable) says the authors seeded liver organoids into drug-screening plates at Day 21, supporting the use of functional organoids at that timepoint. It does not identify Day 21 as an important stage in their functional development or show a developmental change specifically around that day. (LLM, conf=0.82)

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References 55 · Cited by 378

Research works printed but not linked: 39

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Authors
Shinozawa, T., Kimura, M., Cai, Y., Saiki, N., Yoneyama, Y., Ouchi, R., Koike, H., Maezawa, M., Zhang, R.-R., Dunn, A., Ferguson, A., Togo, S., Lewis, K., Thompson, W. L., Asai, A., & Takebe, T.
Affiliations
  • Cincinnati Children's Hospital Medical Center (US)
  • Tokyo Medical and Dental University (JP)
Corresponding author
Takebe, Takanori
ORCID
Venue
Gastroenterology
Publisher
Elsevier BV
Published
2021-02
Volume
160
Issue
3
Pages
831-846.e10
ISSN
0016-5085 (ISSN-L)
DOI
10.1053/j.gastro.2020.10.002
PMC
PMC7878295
OpenAlex
W3092088977
Cited by
378
References
55
Citations by year
2026: 68 · 2025: 91 · 2024: 66 · 2023: 68 · 2022: 50 · 2021: 28 · 2020: 5 · 2012: 1
FWCI
33.37
Type
journal article
Language
en
Open access
Licence
Locations
  • Gastroenterology (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • New York Stem Cell Foundation
  • Cincinnati Children's Hospital Medical Center
  • U.S. Public Health Service (P30 DK078392)
  • Takeda Science Foundation
  • Japan Society for the Promotion of Science (19K22416, JP18H02800)
  • Mitsubishi Foundation
  • Precursory Research for Embryonic Science and Technology (DP2 DK128799-01)
  • Japan Agency for Medical Research and Development (20fk0210060h0002, 20gm1210012h0001, JP19bm0404045, JP19bm0704025, JP19fk0210037, JP19fk0210060)
  • Cincinnati Children’s Research Foundation
Topics
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
  • Health Sciences › Veterinary › Small Animals › Animal testing and alternatives
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
Keywords
drug-induced liver injury, human liver organoids, bile transport, bile salt export pump, hepatocyte zonation, mitochondrial toxicity, drug screening, CYP2C9*2, bosentan
MeSH terms
Cell Line, Drug Evaluation, Preclinical, Humans, Liver, Organoids, Hepatocytes, Toxicity Tests, Acute, Pluripotent Stem Cells, Chemical and Drug Induced Liver Injury, High-Throughput Screening Assays

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