Prediction of liver toxicity and mode of action using metabolomics in vitro in HepG2 cells

Ramı́rez, Tzutzuy; Strigun, Alexander; Verlohner, Andreas; Huener, Hans Albrecht; Peter, Erik; Herold, Michael; Bordag, Natalie; Mellert, W.; Walk, T.; Spitzer, Michael; Jiang, Xiaoqi; Sperber, Saskia; Hofmann, Thomas; Hartung, Thomas; Kamp, Hennicke; Ravenzwaay, B. van

doi:10.1007/s00204-017-2079-6

Cited by 125 publications

(93 citation statements)

References 41 publications

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“…Цитотоксический эффект PLGA-плацебо НЧ оценивали на клеточных линиях HepG2, LLC-PK и U87MG. Культуры клеток LLC-PK и HepG2 часто используют как in vitro модели для оценки нефротоксического и гепатотоксического действия лекар-ственных средств [30,31]. Цитотоксический эффект Dox-PLGA НЧ в отношении опухолевых клеток оценивали на культурах клеток U87MG и HepG2.…”

Section: оценка цитотоксического действия наночастицunclassified

Cytotoxicity and Hemocompatibility of Doxorubicin-Loaded Plga Nanoparticles

Малиновская

Kovalenko

Kovshova

et al. 2020

Rossijskij bioterapevtičeskij žurnal

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Introduction. The use of polymeric biodegradable nanoparticles (NP) as drug delivery systems is a promising approach to overcome histohematomatic barriers. Thus, poloxamer 188-coated poly (lactide-co-glycolide) (PLGA) NP are able to overcome blood-brain barrier and to deliver therapeutic agents, in particular doxorubicin, into intracranial tumour upon intravenous administration. It is important to evaluate NP interaction with blood components in preclinical studies.The objective of the study was to investigate cytotoxicity and hemocompatibility of doxorubicin-loaded PLGA NP (Dox-PLGA NP), to essess NP uptake by glioblastoma cells.Materials and methods. The influence of NP on coagulation cascade was evaluated by prothrombin time measuring before and after plasma incubation with NP. To assess NP thrombogenicity the platelet activation level was determined by flow cytometry. The NP hemolytic activity (released hemoglobin concentration) was measured spectrophotometrically. NP cytotoxicity was determined by MTS assay. NP uptake by human glioblastoma cells was evaluated by flow cytometry.Results. Dox-PLGA NP did not influence blood coagulation time and thrombocyte activity at concentrations up to 100 mcg/mL: PT values were 12–15 s for all tested samples, and P-selectin expression level did not exceed 15 %. All samples were not hemolytic after 3 h of incubation. Cytotoxicity of doxorubicin released from PLGA NP on glioma U87MG cells was comparable to that of free doxorubicin. As shown by flow cytometry Dox-PLGA NP were efficiently internalized into the cells.Conclusion. The study of hemocompatibility confirmed the safety of Dox-PLGA NP: NP did not influence blood coagulation system and did not induce hemolysis. NP were efficiently internalized into the human glioblastoma cells and produced considerable antitumor effect in vitro.

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Section: оценка цитотоксического действия наночастицunclassified

Cytotoxicity and Hemocompatibility of Doxorubicin-Loaded Plga Nanoparticles

Малиновская

Kovalenko

Kovshova

et al. 2020

Rossijskij bioterapevtičeskij žurnal

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“…Other studies HepG2 HepG2, a perpetual hepatocellular cell line, expresses a variety of liver-specific enzymes and nuclear transcription factors (such as p53 and Nrf2) that play pivotal roles in the development of drug-induced toxicity (Jiang et al 2015b). Due to their low cost, high reproducibility and unlimited lifespan, HepG2-based in vitro model systems and multiple omics approaches have been extensively exploited in a number of studies for the prediction of drug-induced hepatotoxicity (Ramirez et al 2018), the classification of cholestatic and necrotic hepatotoxicants (Van den Hof et al 2014) and the discrimination between genotoxicants and non-genotoxicants (Magkoufopoulou et al 2011(Magkoufopoulou et al , 2012 as well as to illustrate the corresponding mechanisms. For instance, an in vitro study conducted using HepG2 cells showed that the transcriptome-based classifiers achieved high accuracies (accuracies above 90%) for the discrimination of hepatotoxicants from nonhepatotoxicants and for the separation of cholestatic and non-cholestatic compounds (36-gene and 12-gene classifiers, respectively) (Van den Hof et al 2014).…”

Section: Hepatocellular Carcinoma Cell Linesmentioning

confidence: 99%

“…Aside from the classification power, this in vitro culture system revealed that endoplasmic reticulum (ER) stress and the unfolded protein response are important cellular responses to drug-induced liver toxicity (Van den Hof et al 2014). In 2018, a research team exposed HepG2 cells to 35 compounds and performed metabolome analyses using mass spectrometry (MS), aiming to develop robust, standardized and reproducible metabolomic systems for prediction of liver toxicity in vitro (Ramirez et al 2018). The study successfully discovered dose-dependent and compound-specific action modes for the tested hepatotoxins and identified several molecular mechanisms (e.g.…”

Section: Hepatocellular Carcinoma Cell Linesmentioning

confidence: 99%

The application of omics-based human liver platforms for investigating the mechanism of drug-induced hepatotoxicity in vitro

et al. 2019

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Drug-induced liver injury (DILI) complicates safety assessment for new drugs and poses major threats to both patient health and drug development in the pharmaceutical industry. A number of human liver cell-based in vitro models combined with toxicogenomics methods have been developed as an alternative to animal testing for studying human DILI mechanisms. In this review, we discuss the in vitro human liver systems and their applications in omics-based drug-induced hepatotoxicity studies. We furthermore present bioinformatic approaches that are useful for analyzing toxicogenomic data generated from these models and discuss their current and potential contributions to the understanding of mechanisms of DILI. Human pluripotent stem cells, carrying donor-specific genetic information, hold great potential for advancing the study of individualspecific toxicological responses. When co-cultured with other liver-derived non-parenchymal cells in a microfluidic device, the resulting dynamic platform enables us to study immune-mediated drug hypersensitivity and accelerates personalized drug toxicology studies. A flexible microfluidic platform would also support the assembly of a more advanced organs-ona-chip device, further bridging gap between in vitro and in vivo conditions. The standard transcriptomic analysis of these cell systems can be complemented with causality-inferring approaches to improve the understanding of DILI mechanisms. These approaches involve statistical techniques capable of elucidating regulatory interactions in parts of these mechanisms. The use of more elaborated human liver models, in harmony with causality-inferring bioinformatic approaches will pave the way for establishing a powerful methodology to systematically assess DILI mechanisms across a wide range of conditions. Keywords Drug-induced hepatotoxicity • In vitro human liver model • Omics approaches • Bioinformatics • Graphical Gaussian networks • Bayesian networks

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“…The human hepatoma (HepG2) cell line was well characterized for its usefulness for examining the cytotoxicity of the substances affecting cell metabolism [26][27][28][29]. The HepG2 cells were selected as target cells for evaluating toxicity to human liver cancer and have been extensively used as the test system for the prediction of toxicity and metabolites in cancer patients [30]. The toxic effect of CAP metabolites was verified in a metabolically inactive cell model, Balb/3T3 fibroblasts.…”

Section: Introductionmentioning

confidence: 99%

Metabolomic Profile of Primary Turkey and Rat Hepatocytes and Two Cell Lines after Chloramphenicol Exposure

Radko

Śniegocki

Sell

et al. 2019

Animals

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Simple Summary: The use of cell cultures can be an important alternative for animal experiments. Therefore, it seems reasonable to use in vitro models to compare the liver metabolism of a drug in different animal species. Chloramphenicol is an effective broad-spectrum antibiotic used in human and pets. The toxicity of chloramphenicol is complex and differs among animal species mainly depending on the biotransformation. The purpose of this study was to assess chloramphenicol metabolism on its cytotoxicity in primary turkey and rat hepatocyte cultures, also in human hepatoma (HepG2) cells and nonhepatic, Balb/c 3T3 fibroblasts. To the best of our knowledge, this is the first report of differences in chloramphenicol metabolism in primary turkey and rat hepatocyte cultures. The two metabolites of the drug were detected in turkey and rat hepatocyte cultures. The amount of one metabolite of chloramphenicol was closely related to the cytotoxicity of the drug. The primary turkey and rat hepatocyte cultures were more sensitive to chloramphenicol than HepG2 cells and Balb/c 3T3 fibroblasts. The primary hepatocyte cultures represent valuable tools with which to study the biotransformation of xenobiotics and determine species differences in their metabolism and toxicity. Abstract:The purpose of this study was to assess the formation of chloramphenicol metabolites in primary turkey and rat hepatocyte cultures and human hepatoma (HepG2) cells and nonhepatic, Balb/c 3T3 fibroblasts. Additionally, the cytotoxicity of the drug was assessed through three biochemical endpoints: mitochondrial and lysosomal activity and cellular membrane integrity after 24 and 48 h exposure. The two metabolites of the drug, chloramphenicol glucuronide and nitroso-chloramphenicol, were detected to the greatest extent in both primary hepatocyte cultures by liquid chromatography-tandem mass spectrometry. Toxic nitroso-chloramphenicol was the main metabolite in the primary turkey hepatocyte cultures, but it was not in the primary rat hepatocyte cultures. The most affected endpoint in turkey and rat hepatocyte cultures was the disintegration of the cellular membrane, but in the cell lines, mitochondrial and lysosomal activities underwent the greatest change. The primary hepatocyte cultures represent valuable tools with which to study the species differences in the biotransformation and toxicity of drugs. To the best of our knowledge, this is the first report of differences in chloramphenicol metabolism in primary turkey and rat hepatocyte cultures.

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Prediction of liver toxicity and mode of action using metabolomics in vitro in HepG2 cells

Cited by 125 publications

References 41 publications

Cytotoxicity and Hemocompatibility of Doxorubicin-Loaded Plga Nanoparticles

Cytotoxicity and Hemocompatibility of Doxorubicin-Loaded Plga Nanoparticles

The application of omics-based human liver platforms for investigating the mechanism of drug-induced hepatotoxicity in vitro

Metabolomic Profile of Primary Turkey and Rat Hepatocytes and Two Cell Lines after Chloramphenicol Exposure

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