Human liver organoids; a patient-derived primary model for HBV Infection and Related Hepatocellular Carcinoma

AbstractThe 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, 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 by 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 disease surveillance and development of HCC in HBV infected patients.

Download Full-text

Application of human liver organoids as a patient-derived primary model for HBV infection and related hepatocellular carcinoma

eLife ◽

10.7554/elife.60747 ◽

2021 ◽

Vol 10 ◽

Author(s):

Elisa De Crignis ◽

Tanvir Hossain ◽

Shahla Romal ◽

Fabrizia Carofiglio ◽

Panagiotis Moulos ◽

...

Keyword(s):

Infected Patient ◽

Human Liver ◽

Ex Vivo ◽

Gene Signature ◽

Hbv Infection ◽

Infection Model ◽

Drug Induced ◽

Molecular Events ◽

Primary Model ◽

Liver Organoids

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.

Download Full-text

3D human liver organoids: An in vitro platform to investigate HBV infection, replication and liver tumorigenesis

Cancer Letters ◽

10.1016/j.canlet.2021.02.024 ◽

2021 ◽

Vol 506 ◽

pp. 35-44

Author(s):

Shringar Rao ◽

Tanvir Hossain ◽

Tokameh Mahmoudi

Keyword(s):

Human Liver ◽

Hbv Infection ◽

Liver Organoids

Download Full-text

SAT-179-Human liver organoids culture as a novel in vitro model of HBV infection

Journal of Hepatology ◽

10.1016/s0618-8278(19)31414-8 ◽

2019 ◽

Vol 70 (1) ◽

pp. e708

Author(s):

Chuan Kok Lim ◽

Bang M Tran ◽

Byron Shue ◽

Nicholas Eyre ◽

Dustin Flanagan ◽

...

Keyword(s):

Human Liver ◽

In Vitro Model ◽

Hbv Infection ◽

Vitro Model ◽

Liver Organoids

Download Full-text

Use of 3D Human Liver Organoids to Predict Drug-Induced Phospholipidosis

International Journal of Molecular Sciences ◽

10.3390/ijms21082982 ◽

2020 ◽

Vol 21 (8) ◽

pp. 2982

Author(s):

Ji-Young Lee ◽

Hyo-Jeong Han ◽

Sang-Joon Lee ◽

Eun-Ho Cho ◽

Han-Byul Lee ◽

...

Keyword(s):

Hepg2 Cells ◽

Human Liver ◽

Three Dimensional ◽

Lamellar Bodies ◽

Albumin Secretion ◽

Drug Induced ◽

Cytoplasmic Vacuoles ◽

Transmission Electron ◽

Liver Organoids

Drug-induced phospholipidosis (PL) is a storage disorder caused by the formation of phospholipid-drug complexes in lysosomes. Because of the diversity of PL between species, human cell-based assays have been used to predict drug-induced PL in humans. We established three-dimensional (3D) human liver organoids as described previously and investigated their liver characteristics through multiple analyses. Drug-induced PL was initiated in these organoids and in monolayer HepG2 cultures, and cellular changes were systemically examined. Organoids that underwent differentiation showed characteristics of hepatocytes rather than HepG2 cells. The organoids also survived under PL-inducing drug conditions for 48 h and maintained a more stable albumin secretion level than the HepG2 cells. More cytoplasmic vacuoles were observed in organoids and HepG2 cells treated with more potent PL-induced drugs, but to a greater extent in organoids than in HepG2 cells. Lysosome-associated membrane protein 2, a marker of lysosome membranes, showed a stronger immunohistochemical signal in the organoids. PL-distinctive lamellar bodies were observed only in amiodarone-treated organoids by transmission electron microscopy. Human liver organoids are thus more sensitive to drug-induced PL and less affected by cytotoxicity than HepG2 cells. Since PL is a chronic condition, these results indicate that organoids better reflect metabolite-mediated hepatotoxicity in vivo and could be a valuable system for evaluating the phospholipidogenic effects of different compounds during drug development.

Download Full-text

Faculty Opinions recommendation of Utility of spherical human liver microtissues for prediction of clinical drug-induced liver injury.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.727717884.793538537 ◽

2017 ◽

Author(s):

Muireann Coen ◽

Rabiya Zia

Keyword(s):

Liver Injury ◽

Human Liver ◽

Drug Induced ◽

Drug Induced Liver Injury ◽

Clinical Drug

Download Full-text

In Vitro, In Silico and Ex Vivo Studies of Dihydroartemisinin Derivatives as Antitubercular Agents

Current Topics in Medicinal Chemistry ◽

10.2174/1568026619666190305131425 ◽

2019 ◽

Vol 19 (8) ◽

pp. 633-644 ◽

Cited By ~ 1

Author(s):

Komal Kalani ◽

Sarfaraz Alam ◽

Vinita Chaturvedi ◽

Shyam Singh ◽

Feroz Khan ◽

...

Keyword(s):

Bone Marrow ◽

In Silico ◽

Ex Vivo ◽

Virulent Strain ◽

Infection Model ◽

Mouse Bone Marrow ◽

Significant Activity ◽

Macrophage Infection ◽

In Silico Studies

Introduction: As a part of our drug discovery program for anti-tubercular agents, dihydroartemisinin (DHA-1) was screened against Mtb H37Rv, which showed moderate anti-tubercular activity (>25.0 µg/mL). These results prompted us to carry out the chemical transformation of DHA-1 into various derivatives and study their antitubercular potential. Materials and Methods: DHA-1 was semi-synthetically converted into four new acyl derivatives (DHA-1A – DHA-1D) and in-vitro evaluated for their anti-tubercular potential against Mycobacterium tuberculosis H37Rv virulent strain. The derivatives, DHA-1C (12-O-(4-nitro) benzoyl; MIC 12.5 µg/mL) and DHA-1D (12-O-chloro acetyl; MIC 3.12µg/mL) showed significant activity against the pathogen. Results: In silico studies of the most active derivative (DHA-1D) showed interaction with ARG448 inhibiting the mycobacterium enzymes. Additionally, it showed no cytotoxicity towards the Vero C1008 cells and Mouse bone marrow derived macrophages. Conclusion: DHA-1D killed 62% intracellular M. tuberculosis in Mouse bone marrow macrophage infection model. To the best of our knowledge, this is the first-ever report on the antitubercular potential of dihydroartemisinin and its derivatives. Since dihydroartemisinin is widely used as an antimalarial drug; these results may be of great help in anti-tubercular drug development from a very common, inexpensive, and non-toxic natural product.

Download Full-text

Targeting human Acyl-CoA:cholesterol acyltransferase as a dual viral and T cell metabolic checkpoint

Nature Communications ◽

10.1038/s41467-021-22967-7 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Nathalie M. Schmidt ◽

Peter A. C. Wing ◽

Mariana O. Diniz ◽

Laura J. Pallett ◽

Leo Swadling ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Human Liver ◽

Lipid Droplets ◽

Ex Vivo ◽

Functional Cure ◽

Carcinogenic Activity ◽

Attractive Therapeutic Target ◽

Tcr Signalling

AbstractDetermining divergent metabolic requirements of T cells, and the viruses and tumours they fail to combat, could provide new therapeutic checkpoints. Inhibition of acyl-CoA:cholesterol acyltransferase (ACAT) has direct anti-carcinogenic activity. Here, we show that ACAT inhibition has antiviral activity against hepatitis B (HBV), as well as boosting protective anti-HBV and anti-hepatocellular carcinoma (HCC) T cells. ACAT inhibition reduces CD8+ T cell neutral lipid droplets and promotes lipid microdomains, enhancing TCR signalling and TCR-independent bioenergetics. Dysfunctional HBV- and HCC-specific T cells are rescued by ACAT inhibitors directly ex vivo from human liver and tumour tissue respectively, including tissue-resident responses. ACAT inhibition enhances in vitro responsiveness of HBV-specific CD8+ T cells to PD-1 blockade and increases the functional avidity of TCR-gene-modified T cells. Finally, ACAT regulates HBV particle genesis in vitro, with inhibitors reducing both virions and subviral particles. Thus, ACAT inhibition provides a paradigm of a metabolic checkpoint able to constrain tumours and viruses but rescue exhausted T cells, rendering it an attractive therapeutic target for the functional cure of HBV and HBV-related HCC.

Download Full-text