Inter-donor variability of phase I/phase II metabolism of three reference drugs in cryopreserved primary human hepatocytes in suspension and monolayer

Braver-Sewradj, Shalenie P. den; Braver, Michiel W. den; Vermeulen, Nico; Commandeur, Jan N. M.; Richert, Lysiane; Vos, J. Chris

doi:10.1016/j.tiv.2016.02.013

Cited by 38 publications

(26 citation statements)

References 66 publications

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“…For the evaluation of drug-induced liver injury (DILI), primary human hepatocytes (PHHs) have been considered the gold standard model ( Gómez-Lechón et al , 2014 ; Sison-Young et al , 2017 ). In conventional 2D monolayer cultures, however, PHH rapidly dedifferentiate within hours after plating and lose hepatic functions, such as albumin production and drug metabolizing enzyme activity ( den Braver-Sewradj et al , 2016 ), which severely limits their predictive capacity, particularly where chemically reactive metabolites are implicated in the toxicity mechanisms ( Heslop et al , 2016 ; Lauschke et al , 2016b ).…”

mentioning

confidence: 99%

Comparison of Hepatic 2D Sandwich Cultures and 3D Spheroids for Long-term Toxicity Applications: A Multicenter Study

Bell

Dankers

Lauschke

et al. 2018

Toxicological Sciences

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Primary human hepatocytes (PHHs) are commonly used for in vitro studies of drug-induced liver injury. However, when cultured as 2D monolayers, PHH lose crucial hepatic functions within hours. This dedifferentiation can be ameliorated when PHHs are cultured in sandwich configuration (2Dsw), particularly when cultures are regularly re-overlaid with extracellular matrix, or as 3D spheroids. In this study, the 6 participating laboratories evaluated the robustness of these 2 model systems made from cryopreserved PHH from the same donors considering both inter-donor and inter-laboratory variability and compared their suitability for use in repeated-dose toxicity studies using 5 different hepatotoxins with different toxicity mechanisms. We found that expression levels of proteins involved in drug absorption, distribution, metabolism, and excretion, as well as catalytic activities of 5 different CYPs, were significantly higher in 3D spheroid cultures, potentially affecting the exposure of the cells to drugs and their metabolites. Furthermore, global proteomic analyses revealed that PHH in 3D spheroid configuration were temporally stable whereas proteomes from the same donors in 2Dsw cultures showed substantial alterations in protein expression patterns over the 14 days in culture. Overall, spheroid cultures were more sensitive to the hepatotoxic compounds investigated, particularly upon long-term exposures, across testing sites with little inter-laboratory or inter-donor variability. The data presented here suggest that repeated-dosing regimens improve the predictivity of in vitro toxicity assays, and that PHH spheroids provide a sensitive and robust system for long-term mechanistic studies of drug-induced hepatotoxicity, whereas the 2Dsw system has a more dedifferentiated phenotype and lower sensitivity to detect hepatotoxicity.

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mentioning

confidence: 99%

Comparison of Hepatic 2D Sandwich Cultures and 3D Spheroids for Long-term Toxicity Applications: A Multicenter Study

Bell

Dankers

Lauschke

et al. 2018

Toxicological Sciences

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241

203

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“…Several publications have recently appeared in which the here selected training compounds and negative controls have been investigated in order to substantiate their utility as training compounds to profile and elucidate further DILI mechanisms and to interrogate existing and novel in vitro systems for the prediction of human DILI (Sison-Young et al 2015; Bell et al 2016; Sison-Young et al 2016; den Braver-Sewradj et al 2016; den Braver et al 2016). …”

Section: Discussionmentioning

confidence: 99%

Evidence-based selection of training compounds for use in the mechanism-based integrated prediction of drug-induced liver injury in man

Dragović¹,

Vermeulen²,

Gerets

et al. 2016

Arch Toxicol

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The current test systems employed by pharmaceutical industry are poorly predictive for drug-induced liver injury (DILI). The ‘MIP-DILI’ project addresses this situation by the development of innovative preclinical test systems which are both mechanism-based and of physiological, pharmacological and pathological relevance to DILI in humans. An iterative, tiered approach with respect to test compounds, test systems, bioanalysis and systems analysis is adopted to evaluate existing models and develop new models that can provide validated test systems with respect to the prediction of specific forms of DILI and further elucidation of mechanisms. An essential component of this effort is the choice of compound training set that will be used to inform refinement and/or development of new model systems that allow prediction based on knowledge of mechanisms, in a tiered fashion. In this review, we focus on the selection of MIP-DILI training compounds for mechanism-based evaluation of non-clinical prediction of DILI. The selected compounds address both hepatocellular and cholestatic DILI patterns in man, covering a broad range of pharmacologies and chemistries, and taking into account available data on potential DILI mechanisms (e.g. mitochondrial injury, reactive metabolites, biliary transport inhibition, and immune responses). Known mechanisms by which these compounds are believed to cause liver injury have been described, where many if not all drugs in this review appear to exhibit multiple toxicological mechanisms. Thus, the training compounds selection offered a valuable tool to profile DILI mechanisms and to interrogate existing and novel in vitro systems for the prediction of human DILI.

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“…The reaction of drug metabolism can be divided into two categories: phase I and phase II; CYPs play an important role in the metabolism of phase I metabolism and the pharmacokinetics of xenobiotics. The meaningful processes observed in phase I include hydrolysis, oxidation, and reduction [12,13]. The second period is mainly directed toward generally lipophilic compounds, and occurs by conjugation with endogenous substances such as glutathione (GSH), glucuronic acid, and sulfate [14].…”

Section: F I G U R Ementioning

confidence: 99%

Electrochemical microreactor combined with mass spectrometry for online oxidation and real‐time detection of alkaloids

Yang

Wang³

et al. 2020

J of Separation Science

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The main purpose of the present study was to investigate the prototypes and oxidation products of alkaloids with the use of an online electrochemistry/quadrupole time-of-flight mass spectrometry system. The metabolism of oxidative phase I and II was simulated in an electrochemical reaction cell. The metabolic processes for coptisine and jatrorrhizine were simulated in a thin-layer cell fitted with a glassy carbon working electrode, while the metabolic processes for berberine and palmatine were simulated by using a boron-doped diamond working electrode. By using the new experimental system, dehydrogenation, demethylation, methylation, hydroxylation, and the formation of two hydroxylation adducts were detected by applying different potentials to the electrochemical cell. The online reaction with glutathione yielded different covalent glutathione adducts. The results obtained from the electrochemical simulation were found to be in good accordance with those reported previously in vivo, showing that electrochemistry/mass spectrometry is an effective tool for studying metabolic reactions for various complex components. Moreover, analysis of alkaloids in liver microsomes by liquid chromatography coupled with mass spectrometry confirmed the possibility of using an electrochemistry technique to simulate the metabolism of target compounds.

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Inter-donor variability of phase I/phase II metabolism of three reference drugs in cryopreserved primary human hepatocytes in suspension and monolayer

Cited by 38 publications

References 66 publications

Comparison of Hepatic 2D Sandwich Cultures and 3D Spheroids for Long-term Toxicity Applications: A Multicenter Study

Comparison of Hepatic 2D Sandwich Cultures and 3D Spheroids for Long-term Toxicity Applications: A Multicenter Study

Evidence-based selection of training compounds for use in the mechanism-based integrated prediction of drug-induced liver injury in man

Electrochemical microreactor combined with mass spectrometry for online oxidation and real‐time detection of alkaloids

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