Perfused human hepatocyte microtissues identify reactive metabolite-forming and mitochondria-perturbing hepatotoxins

Rowe, Cliff; Shaeri, Mohsen; Large, Emma; Cornforth, Terri; Robinson, Angela Byun; Kostrzewski, Tomasz; Sison-Young, Rowena; Goldring, Christopher E.; Park, Kevin; Hughes, David J.

doi:10.1016/j.tiv.2017.09.012

Cited by 26 publications

(18 citation statements)

References 44 publications

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“…Cells in this model remain viable and functional for at least seven days, as judged by calcein acetoxymethyl (AM) and albumin staining, respectively. The model has been used for a variety of applications, including pharmacokinetic studies, evaluation of hepatotoxicity, modeling of nonalcoholic fatty liver disease (NAFLD), and hepatitis B viral infections . In addition, a multitude of small companies and start‐ups provide liver‐on‐a‐chip platforms for which at present very limited peer‐reviewed benchmarking data are available.…”

Section: D Culture Paradigms For Phhmentioning

confidence: 99%

3D Primary Hepatocyte Culture Systems for Analyses of Liver Diseases, Drug Metabolism, and Toxicity: Emerging Culture Paradigms and Applications

Lauschke

Shafagh

Hendriks

et al. 2019

Biotechnology Journal

101

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Recent research has shown that the maintenance of relevant liver functions ex vivo requires models in which the cells exhibit an in vivo‐like phenotype, often achieved by reconstitution of appropriate cellular interactions. Multiple different models have been presented that differ in the cells utilized, media, and culture conditions. Furthermore, several technologically different approaches have been presented including bioreactors, chips, and plate‐based systems in fluidic or static media constituting of chemically diverse materials. Using such models, the ability to predict drug metabolism, drug toxicity, and liver functionality have increased tremendously as compared to conventional in vitro models in which cells are cultured as 2D monolayers. Here, the authors highlight important considerations for microphysiological systems for primary hepatocyte culture, review current culture paradigms, and discuss their opportunities for studies of drug metabolism, hepatotoxicity, liver biology, and disease.

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Section: D Culture Paradigms For Phhmentioning

confidence: 99%

3D Primary Hepatocyte Culture Systems for Analyses of Liver Diseases, Drug Metabolism, and Toxicity: Emerging Culture Paradigms and Applications

Lauschke

Shafagh

Hendriks

et al. 2019

Biotechnology Journal

101

View full text Add to dashboard Cite

show abstract

“…However, expression levels of various CYPs (CYP2C8 and CYP2E1), drug transporters (BSEP and OATP1B3), and nuclear receptors (CAR and PXR) decline considerably over the course of 10 days (Vivares et al, 2014). Nevertheless, hierarchical clustering of lactate dehydrogenase (LDH) leakage, albumin secretion and urea production distinguished the hepatotoxins fialuridine, acetaminophen, and clozapine from the nontoxins olanzapine and entecavir (Rowe et al, 2018). In a co-culture configuration of PHH and Kupffer cells, the platform remained metabolically stable for 2 weeks and could model the modulation of CYP3A4 activity by the monoclonal antibody tocilizumab and its effects on simvastatin hydroxy acid pharmacokinetics (Long et al, 2016).…”

Section: Organotypic Liver Models For Hepatotoxicity Predictionsmentioning

confidence: 99%

Comprehensive Evaluation of Organotypic and Microphysiological Liver Models for Prediction of Drug-Induced Liver Injury

2019

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Drug-induced liver injury (DILI) is a major concern for the pharmaceutical industry and constitutes one of the most important reasons for the termination of promising drug development projects. Reliable prediction of DILI liability in preclinical stages is difficult, as current experimental model systems do not accurately reflect the molecular phenotype and functionality of the human liver. As a result, multiple drugs that passed preclinical safety evaluations failed due to liver toxicity in clinical trials or postmarketing stages in recent years. To improve the selection of molecules that are taken forward into the clinics, the development of more predictive in vitro systems that enable high-throughput screening of hepatotoxic liabilities and allow for investigative studies into DILI mechanisms has gained growing interest. Specifically, it became increasingly clear that the choice of cell types and culture method both constitute important parameters that affect the predictive power of test systems. In this review, we present current 3D culture paradigms for hepatotoxicity tests and critically evaluate their utility and performance for DILI prediction. In addition, we highlight possibilities of these emerging platforms for mechanistic evaluations of selected drug candidates and present current research directions towards the further improvement of preclinical liver safety tests. We conclude that organotypic and microphysiological liver systems have provided an important step towards more reliable DILI prediction. Furthermore, we expect that the increasing availability of comprehensive benchmarking studies will facilitate model dissemination that might eventually result in their regulatory acceptance.

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“…(20) We previously developed a model of hepatic steatosis using a 3D perfused microphysiological system (MPS), which enabled PHHs to be cultured for 2 weeks in the presence of FFAs, allowing the chronic effects of triglyceride accumulation to be analyzed. (21) The perfused MPS maintains highly metabolically active PHHs for extended periods (up to 40 days) (22)(23)(24) and has been demonstrated to support PHHs and human Kupffer cell (HK) co-cultures to study the effects of liver inflammation on drug metabolism, drug-drug interactions, and liver toxicity. (22)(23)(24) Here, we use the same MPS with a co-culture of PHH, HK, and hepatic stellate cells (HSCs) to create a model of human NASH.…”

Section: A Microphysiological System For Studying Nonalcoholic Steatomentioning

confidence: 99%

“…(21) The perfused MPS maintains highly metabolically active PHHs for extended periods (up to 40 days) (22)(23)(24) and has been demonstrated to support PHHs and human Kupffer cell (HK) co-cultures to study the effects of liver inflammation on drug metabolism, drug-drug interactions, and liver toxicity. (22)(23)(24) Here, we use the same MPS with a co-culture of PHH, HK, and hepatic stellate cells (HSCs) to create a model of human NASH. These co-cultures can be maintained under disease-inducing conditions for at least 2 weeks and demonstrate key hallmarks of the human disease, including accumulation of intracellular triglyceride, the production of both pro-inflammatory cytokines, and profibrotic markers.…”

Section: A Microphysiological System For Studying Nonalcoholic Steatomentioning

confidence: 99%

A Microphysiological System for Studying Nonalcoholic Steatohepatitis

et al. 2019

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Nonalcoholic steatohepatitis (NASH) is the most severe form of nonalcoholic fatty liver disease (NAFLD), which to date has no approved drug treatments. There is an urgent need for better understanding of the genetic and molecular pathways that underlie NAFLD/NASH, and currently available preclinical models, be they in vivo or in vitro, do not fully represent key aspects of the human disease state. We have developed a human in vitro co‐culture NASH model using primary human hepatocytes, Kupffer cells and hepatic stellate cells, which are cultured together as microtissues in a perfused three‐dimensional microphysiological system (MPS). The microtissues were cultured in medium containing free fatty acids for at least 2 weeks, to induce a NASH‐like phenotype. The co‐culture microtissues within the MPS display a NASH‐like phenotype, showing key features of the disease including hepatic fat accumulation, the production of an inflammatory milieu, and the expression of profibrotic markers. Addition of lipopolysaccharide resulted in a more pro‐inflammatory milieu. In the model, obeticholic acid ameliorated the NASH phenotype. Microtissues were formed from both wild‐type and patatin‐like phospholipase domain containing 3 (PNPLA3) I148M mutant hepatic stellate cells. Stellate cells carrying the mutation enhanced the overall disease state of the model and in particular produced a more pro‐inflammatory milieu. Conclusion: The MPS model displays a phenotype akin to advanced NAFLD or NASH and has utility as a tool for exploring mechanisms underlying the disease. Furthermore, we demonstrate that in co‐culture the PNPLA3 I148M mutation alone can cause hepatic stellate cells to enhance the overall NASH disease phenotype.

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Perfused human hepatocyte microtissues identify reactive metabolite-forming and mitochondria-perturbing hepatotoxins

Cited by 26 publications

References 44 publications

3D Primary Hepatocyte Culture Systems for Analyses of Liver Diseases, Drug Metabolism, and Toxicity: Emerging Culture Paradigms and Applications

3D Primary Hepatocyte Culture Systems for Analyses of Liver Diseases, Drug Metabolism, and Toxicity: Emerging Culture Paradigms and Applications

Comprehensive Evaluation of Organotypic and Microphysiological Liver Models for Prediction of Drug-Induced Liver Injury

A Microphysiological System for Studying Nonalcoholic Steatohepatitis

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