scholarly journals Primary Human Hepatocyte Spheroids as Tools to Study the Hepatotoxic Potential of Non-Pharmaceutical Chemicals

2021 ◽  
Vol 22 (20) ◽  
pp. 11005
Author(s):  
Vânia Vilas-Boas ◽  
Eva Gijbels ◽  
Kaat Leroy ◽  
Alanah Pieters ◽  
Audrey Baze ◽  
...  
Keyword(s):  
Cyclosporine A ◽  
Prolonged Exposure ◽  
Human Hepatocyte ◽  
Drug Induced ◽  
Clinical Issue ◽  
Drug Induced Liver Injury ◽  
Primary Human Hepatocyte ◽  
Donor Variability ◽  
Hepatocyte Spheroids

Drug-induced liver injury, including cholestasis, is an important clinical issue and economic burden for pharmaceutical industry and healthcare systems. However, human-relevant in vitro information on the ability of other types of chemicals to induce cholestatic hepatotoxicity is lacking. This work aimed at investigating the cholestatic potential of non-pharmaceutical chemicals using primary human hepatocytes cultured in 3D spheroids. Spheroid cultures were repeatedly (co-) exposed to drugs (cyclosporine-A, bosentan, macitentan) or non-pharmaceutical chemicals (paraquat, tartrazine, triclosan) and a concentrated mixture of bile acids for 4 weeks. Cell viability (adenosine triphosphate content) was checked every week and used to calculate the cholestatic index, an indicator of cholestatic liability. Microarray analysis was performed at specific time-points to verify the deregulation of genes related to cholestasis, steatosis and fibrosis. Despite the evident inter-donor variability, shorter exposures to cyclosporine-A consistently produced cholestatic index values below 0.80 with transcriptomic data partially supporting its cholestatic burden. Bosentan confirmed to be hepatotoxic, while macitentan was not toxic in the tested concentrations. Prolonged exposure to paraquat suggested fibrotic potential, while triclosan markedly deregulated genes involved in different types of hepatotoxicity. These results support the applicability of primary human hepatocyte spheroids to study hepatotoxicity of non-pharmaceutical chemicals in vitro.

scholarly journals Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Catherine C. Bell ◽  
Delilah F. G. Hendriks ◽  
Sabrina M. L. Moro ◽  
Ewa Ellis ◽  
Joanne Walsh ◽  
...  
Keyword(s):  
Liver Injury ◽  
Liver Function ◽  
Model System ◽  
Human Hepatocyte ◽  
Drug Induced ◽  
Drug Induced Liver Injury ◽  
Primary Human Hepatocyte ◽  
Hepatocyte Spheroids

scholarly journals Intermittent Starvation Extends the Functional Lifetime of Primary Human Hepatocyte Cultures

2020 ◽  
Vol 174 (2) ◽  
pp. 266-277
Author(s):  
Matthew D Davidson ◽  
Salman R Khetani
Keyword(s):  
Drug Exposure ◽  
Adenosine Monophosphate ◽  
Human Hepatocyte ◽  
Toxicity Tests ◽  
Drug Induced ◽  
Nonparenchymal Cells ◽  
Primary Human Hepatocyte ◽  
Serum Hormone

Abstract Primary human hepatocyte (PHH) cultures have become indispensable to mitigate the risk of adverse drug reactions in human patients. In contrast to dedifferentiating monocultures, coculture with nonparenchymal cells maintains PHH functions for 2–4 weeks. However, because the functional lifespan of PHHs in vivo is 200–400 days, it is desirable to further prolong PHH functions in vitro toward modeling chronic drug exposure and disease progression. Fasting has benefits on the longevity of organisms and the health of tissues such as the liver. We hypothesized that a culturing protocol that mimics dynamic fasting/starvation could activate starvation pathways and prolong PHH functional lifetime. To mimic starvation, serum and hormones were intermittently removed from the culture medium of micropatterned cocultures (MPCCs) containing PHHs organized onto collagen domains and surrounded by 3T3-J2 murine fibroblasts. A weekly 2-day starvation optimally prolonged PHH functional lifetime for 6+ weeks in MPCCs versus a decline after 3 weeks in nonstarved controls. The 2-day starvation also enhanced the functions of PHH monocultures for 2 weeks, suggesting direct effects on PHHs. In MPCCs, starvation activated 5' adenosine monophosphate-activated protein kinase (AMPK) and restricted fibroblast overgrowth onto PHH islands, thereby maintaining hepatic polarity. The effects of starvation on MPCCs were partially recapitulated by activating AMPK using metformin or growth arresting fibroblasts via mitomycin-C. Lastly, starved MPCCs demonstrated lower false positives for drug toxicity tests and higher drug-induced cytochrome-P450 activities versus nonstarved controls even after 5 weeks. In conclusion, intermittent serum/hormone starvation extends PHH functional lifetime toward enabling clinically relevant drug screening.

scholarly journals Primary human hepatocyte spheroids as an in vitro tool for investigating drug compounds with low clearance

2021 ◽  
pp. DMD-AR-2020-000340
Author(s):  
Julia Riede ◽  
Birgit M. Wollmann ◽  
Espen Molden ◽  
Magnus Ingelman-Sundberg
Keyword(s):  
Human Hepatocyte ◽  
Primary Human Hepatocyte ◽  
Drug Compounds ◽  
Hepatocyte Spheroids

scholarly journals Intermittent Starvation Extends the Functional Lifetime of Primary Human Hepatocyte Cultures

2019 ◽  
Author(s):  
Matthew D. Davidson ◽  
Salman R. Khetani
Keyword(s):  
Protein Kinase ◽  
Drug Exposure ◽  
Adenosine Monophosphate ◽  
Human Hepatocyte ◽  
Toxicity Tests ◽  
Drug Induced ◽  
Primary Human Hepatocyte ◽  
Serum Hormone

Primary human hepatocyte (PHH) cultures have become indispensable to mitigate the risk of adverse drug reactions in human patients. In contrast to de-differentiating monocultures, co-culture with non-parenchymal cells maintains PHH functions for 2-4 weeks. However, since the functional lifespan of PHHs in vivo is 200-400 days, it is desirable to further prolong PHH functions in vitro towards modeling chronic drug exposure and disease progression. Fasting has benefits on the longevity of organisms and the health of tissues such as the liver. We hypothesized that a culturing protocol that mimics dynamic fasting/starvation could activate starvation pathways and prolong PHH functional lifetime. To mimic starvation, serum and hormones were intermittently removed from the culture medium of micropatterned co-cultures (MPCC) containing PHHs organized onto collagen domains and surrounded by 3T3-J2 murine fibroblasts. A weekly 2-day starvation optimally prolonged PHH functional lifetime for 6+ weeks in MPCCs versus a decline after 3 weeks in non-starved controls. The 2-day starvation also enhanced the functions of PHH monocultures for 2 weeks, suggesting direct effects on PHHs. In MPCCs, starvation activated adenosine monophosphate activated protein kinase and restricted fibroblast overgrowth onto PHH islands, thereby maintaining hepatic polarity. The effects of starvation on MPCCs were partially recapitulated by activating adenosine monophosphate activated protein kinase using metformin or growth-arresting fibroblasts via mitomycin-C. Lastly, starved MPCCs demonstrated lower false positives for drug toxicity tests and higher drug-induced cytochrome-P450 activities versus non-starved controls even after 5 weeks. In conclusion, intermittent serum/hormone starvation extends PHH functional lifetime towards enabling clinically-relevant drug screening.

Recent Progress in Prediction Systems for Drug-induced Liver Injury Using in vitro Cell Culture

2020 ◽  
Vol 14 ◽  
Author(s):  
Shogo Ozawa ◽  
Toshitaka Miura ◽  
Jun Terashima ◽  
Wataru Habano ◽  
Seiichi Ishida
Keyword(s):  
Cell Culture ◽  
Recent Progress ◽  
Inflammatory Cells ◽  
Protein Adducts ◽  
In Vitro Assays ◽  
Drug Induced ◽  
Drug Induced Liver Injury ◽  
Culture Systems ◽  
Cell Culture Systems

Background: In order to avoid drug-induced liver injury (DILI), in vitro assays, which enable the assessment of both metabolic activation and immune reaction processes that ultimately result in DILI, are needed. Objective: In this study, the recent progress in the application of in vitro assays using cell culture systems is reviewed for potential DILI-causing drugs/xenobiotics and a mechanistic study on DILI, as well as for the limitations of in vitro cell culture systems for DILI research. Methods: Information related to DILI was collected through a literature search of the PubMed database. Results: The initial biological event for the onset of DILI is the formation of cellular protein adducts after drugs have been metabolically activated by drug metabolizing enzymes. The damaged peptides derived from protein adducts lead to the activation of CD4+ helper T lymphocytes and recognition by CD8+ cytotoxic T lymphocytes, which destroy hepatocytes through immunological reactions. Because DILI is a major cause of drug attrition and drug withdrawal, numerous in vitro systems consisting of hepatocytes and immune/inflammatory cells, or spheroids of human primary hepatocytes containing non-parenchymal cells have been developed. These cellular-based systems have identified DILIinducing drugs with approximately 50% sensitivity and 90% specificity. Conclusion: Different co-culture systems consisting of human hepatocyte-derived cells and other immune/inflammatory cells have enabled the identification of DILI-causing drugs and of the actual mechanisms of action.

Drug-induced liver injury classification model based on in vitro human transcriptomics and in vivo rat clinical chemistry data

2014 ◽  
Vol 2 (4) ◽  
pp. 63-70 ◽  
Author(s):  
Danyel Jennen ◽  
Jan Polman ◽  
Mark Bessem ◽  
Maarten Coonen ◽  
Joost van Delft ◽  
...  
Keyword(s):  
Liver Injury ◽  
Clinical Chemistry ◽  
Classification Model ◽  
Drug Induced ◽  
Drug Induced Liver Injury ◽  
Chemistry Data ◽  
Model Based ◽  
Injury Classification

scholarly journals Computational Models Using Multiple Machine Learning Algorithms for Predicting Drug Hepatotoxicity with the DILIrank Dataset

2020 ◽  
Author(s):  
Robert Ancuceanu ◽  
Marilena Viorica Hovanet ◽  
Adriana Iuliana Anghel ◽  
Florentina Furtunescu ◽  
Monica Neagu ◽  
...  
Keyword(s):  
Machine Learning ◽  
Liver Injury ◽  
Computational Models ◽  
Liver Toxicity ◽  
Learning Algorithms ◽  
Machine Learning Algorithms ◽  
Drug Induced ◽  
Reference Drug ◽  
Drug Induced Liver Injury

Drug induced liver injury (DILI) remains one of the challenges in the safety profile of both authorized drugs and candidate drugs and predicting hepatotoxicity from the chemical structure of a substance remains a challenge worth pursuing, being also coherent with the current tendency for replacing non-clinical tests with in vitro or in silico alternatives. In 2016 a group of researchers from FDA published an improved annotated list of drugs with respect to their DILI risk, constituting “the largest reference drug list ranked by the risk for developing drug-induced liver injury in humans”, DILIrank. This paper is one of the few attempting to predict liver toxicity using the DILIrank dataset. Molecular descriptors were computed with the Dragon 7.0 software, and a variety of feature selection and machine learning algorithms were implemented in the R computing environment. Nested (double) cross-validation was used to externally validate the models selected. A number of 78 models with reasonable performance have been selected and stacked through several approaches, including the building of multiple meta-models. The performance of the stacked models was slightly superior to other models published. The models were applied in a virtual screening exercise on over 100,000 compounds from the ZINC database and about 20% of them were predicted to be non-hepatotoxic.

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