Application of human liver organoids as a patient-derived primary model for HBV infection and related hepatocellular carcinoma
- Erasmus MC, Netherlands
- Biomedical Sciences Research Center 'Alexander Fleming', Greece
- Erasmus University Medical Center, Netherlands
- Foundation Hubrecht Organoid Technology (HUB), Netherlands
- HybridStat Predictive Analytics, Greece
- University Medical Centre Utrecht, Netherlands
- University of Cambridge, United Kingdom
Abstract
The molecular events that drive Hepatitis B virus (HBV)-mediated transformation and tumorigenesis have remained largely unclear, due to the absence of a relevant primary model system. Here we propose the use of human liver organoids as a platform for modeling HBV infection and related tumorigenesis. We first describe a primary ex vivo HBV-infection model derived from healthy donor liver organoids after challenge with recombinant virus or HBV-infected patient serum. HBV infected organoids produced cccDNA, HBeAg, expressed intracellular HBV RNA and proteins, and produced infectious HBV. This ex vivo HBV infected primary differentiated hepatocyte organoid platform was amenable to drug screening for both anti-HBV activity as well as for drug-induced toxicity. We also studied HBV replication in transgenically modified organoids; liver organoids exogenously overexpressing the HBV receptor NTCP after lentiviral transduction were not more susceptible to HBV, suggesting the necessity for additional host factors for efficient infection. We also generated transgenic organoids harboring integrated HBV, representing a long-term culture system also suitable for viral production and the study of HBV transcription. Finally, we generated HBV-infected patient-derived liver organoids from non-tumor cirrhotic tissue of explants from liver transplant patients. Interestingly, transcriptomic analysis of patient-derived liver organoids indicated the presence of an aberrant early cancer gene signature, which clustered with the HCC cohort on the TCGA LIHC dataset and away from healthy liver tissue, and may provide invaluable novel biomarkers for the development of HCC and surveillance in HBV infected patients.
Data availability
Sequencing data that support the findings of this study have been deposited in GEO with the accession code GSE 126798.
Article and author information
Author details
Mir Mubashir Khalid
Funding
European Research Council ERC (STG 337116)
- Tokameh Mahmoudi
Dutch Aids Fonds (201614)
- Tokameh Mahmoudi
Bristol Meyers Squibb (AI424-543)
- Elisa De Crignis
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Melanie Ott, University of California, San Francisco, United States
Ethics
Human subjects: The Medical Ethical Council of the Erasmus Medical Center approved the use of this material for research purposes (reference number: MEC-2014-060, Department of Surgery, Erasmus MC), and informed consent was provided from all patients.
Version history
- Preprint posted: March 5, 2019 (view preprint)
- Received: August 25, 2020
- Accepted: July 29, 2021
- Accepted Manuscript published: July 30, 2021 (version 1)
- Version of Record published: August 24, 2021 (version 2)
Copyright
© 2021, De Crignis et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Application of human liver organoids as a patient-derived primary model for HBV infection and related hepatocellular carcinoma
eLife 10:e60747.
https://doi.org/10.7554/eLife.60747
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