Establishment of a primary human hepatocyte spheroid system for evaluating metabolic toxicity using dacarbazine under conditions of CYP1A2 induction

Mizoi, Kenta; Hosono, Mayu; Kojima, Hajime; Ogihara, Takuo

doi:10.1016/j.dmpk.2019.11.002

Cited by 13 publications

(11 citation statements)

References 33 publications

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“…Many attempts were made to use pHHs in 3D culture with the aim to maintain and to prolong their viability and to prevent dedifferentiation. PHH spheroids were viable for at least 2 and up to 7 weeks with stable albumin production [ 50 , 51 , 52 , 53 , 54 ], urea synthesis [ 51 , 52 ] and glycogen storage [ 54 ]. Additionally, cellular polarization was clearly present in pHH spheroids as shown by MRP2 [ 51 , 53 ], P-glycoprotein (Pgp), [ 54 ], and BSEP expression [ 53 , 54 ].…”

Section: Three-dimensional Culture Models For Human Hepatocytesmentioning

confidence: 99%

Three-Dimensional Liver Culture Systems to Maintain Primary Hepatic Properties for Toxicological Analysis In Vitro

Kammerer

2021

IJMS

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Drug-induced liver injury (DILI) is the major reason for failures in drug development and withdrawal of approved drugs from the market. Two-dimensional cultures of hepatocytes often fail to reliably predict DILI: hepatoma cell lines such as HepG2 do not reflect important primary-like hepatic properties and primary human hepatocytes (pHHs) dedifferentiate quickly in vitro and are, therefore, not suitable for long-term toxicity studies. More predictive liver in vitro models are urgently required in drug development and compound safety evaluation. This review discusses available human hepatic cell types for in vitro toxicology analysis and their usage in established and emerging three-dimensional (3D) culture systems. Generally, 3D cultures maintain or improve primary hepatic functions (including expression of drug-metabolizing enzymes) of different liver cells for several weeks of culture, thus allowing long-term and repeated-dose toxicity studies. Spheroid cultures of pHHs have been comprehensively tested, but also other cell types such as HepaRG benefit from 3D culture systems. Emerging 3D culture techniques include usage of induced pluripotent stem-cell-derived hepatocytes and primary-like upcyte cells, as well as advanced culture techniques such as microfluidic liver-on-a-chip models. In-depth characterization of existing and emerging 3D hepatocyte technologies is indispensable for successful implementation of such systems in toxicological analysis.

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Section: Three-dimensional Culture Models For Human Hepatocytesmentioning

confidence: 99%

Three-Dimensional Liver Culture Systems to Maintain Primary Hepatic Properties for Toxicological Analysis In Vitro

Kammerer

2021

IJMS

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“…Due to their increased metabolic activity, PHH spheroids are particularly suitable to evaluate toxicity caused by drug‐drug interactions. For instance, the proton‐pump inhibitor omeprazole, a known activator of the aryl hydrocarbon receptor (AhR), induced CYP1A2 activity in PHH spheroids, which in turn resulted in increased metabolic activation and elevated hepatotoxicity of the CYP1A2 substrate dacarbazine 60 . Using exposure at sub‐toxic concentrations and transcriptomic analyses, gene alterations could be identified that were specific for different classes of toxicity mechanisms, including amiodarone for mitochondrial toxicity, chlorpromazine for cholestasis and aflatoxin B1 for genotoxicity 61 .…”

Section: Liver Spheroid Applications In Drug Discovery and Developmentmentioning

confidence: 99%

3D human liver spheroids for translational pharmacology and toxicology

Ingelman‐Sundberg

Lauschke

2021

Basic Clin Pharma Tox

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Drug development is a failure‐prone endeavour, and more than 85% of drugs fail during clinical development, showcasing that current preclinical systems for compound selection are clearly inadequate. Liver toxicity remains a major reason for safety failures. Furthermore, all efforts to develop pharmacological therapies for a variety of chronic liver diseases, such as non‐alcoholic steatohepatitis (NASH) and fibrosis, remain unsuccessful. Considering the time and expense of clinical trials, as well as the substantial burden on patients, new strategies are thus of paramount importance to increase clinical success rates. To this end, human liver spheroids are becoming increasingly utilized as they allow to preserve patient‐specific phenotypes and functions for multiple weeks in culture. We here review the recent application of such systems for i) predictive and mechanistic analyses of drug hepatotoxicity, ii) the evaluation of hepatic disposition and metabolite formation of low clearance drugs and iii) the development of drugs for metabolic and infectious liver diseases, including NASH, fibrosis, malaria and viral hepatitis. We envision that with increasing dissemination, liver spheroids might become the new gold standard for such applications in translational pharmacology and toxicology.

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“…Therefore, it is important to assess the ability of compounds to cause DILI in the presence of CYP inducers. We developed a CYP1A2-mediated metabolic toxicity system using PHH spheroids treated with OPZ as a CYP1A2 inducer and dacarbazine (DTIC) as a test substrate [ 56 ], since CYP1A2 catalyzes the formation of a cytotoxic product (reactive oxygen species) from DTIC [ 75 ]. Cell viability, CYP1A2 mRNA expression level, and DTIC metabolism were measured alongside hepatic function markers (albumin and urea secretion and AST leakage).…”

Section: Application Of Phh Spheroids For Evaluating Metabolic Toxmentioning

confidence: 99%

“…Cultured human cancer-derived hepatocytes have long been used for human-specific toxicity assessments, but they generally have lower expression levels of drug-metabolizing enzymes than PHHs [ 39 , 40 ]. Various applications of PHH spheroids as pharmacokinetic models [ 41 , 42 , 43 , 44 ] or hepatotoxicity detection models [ 16 , 45 , 46 , 47 , 48 , 49 , 50 , 51 ] have already been reported, and it has become clear that this model is extremely versatile and offers a variety of advantages over 2D cultures ( Table 2 ) [ 16 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 ]. PHH spheroids have also been used as disease models [ 58 , 59 , 60 ].…”

Section: Introductionmentioning

confidence: 99%

Utility of Three-Dimensional Cultures of Primary Human Hepatocytes (Spheroids) as Pharmacokinetic Models

et al. 2020

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This paper reviews the usefulness, current status, and potential of primary human hepatocytes (PHHs) in three-dimensional (3D) cultures, also known as spheroids, in the field of pharmacokinetics (PK). Predicting PK and toxicity means pharmaceutical research can be conducted more efficiently. Various in vitro test systems using human hepatocytes have been proposed as tools to detect hepatic toxicity at an early stage in the drug development process. However, such evaluation requires long-term, low-level exposure to the test compound, and conventional screening systems such as PHHs in planar (2D) culture, in which the cells can only survive for a few days, are unsuitable for this purpose. In contrast, spheroids consisting of PHH are reported to retain the functional characteristics of human liver for at least 35 days. Here, we introduce a fundamental PK and toxicity assessment model of PHH spheroids and describe their applications for assessing species-specific metabolism, enzyme induction, and toxicity, focusing on our own work in these areas. The studies outlined in this paper may provide important information for pharmaceutical companies to reduce termination of development of drug candidates.

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Establishment of a primary human hepatocyte spheroid system for evaluating metabolic toxicity using dacarbazine under conditions of CYP1A2 induction

Cited by 13 publications

References 33 publications

Three-Dimensional Liver Culture Systems to Maintain Primary Hepatic Properties for Toxicological Analysis In Vitro

Three-Dimensional Liver Culture Systems to Maintain Primary Hepatic Properties for Toxicological Analysis In Vitro

3D human liver spheroids for translational pharmacology and toxicology

Utility of Three-Dimensional Cultures of Primary Human Hepatocytes (Spheroids) as Pharmacokinetic Models

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