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Toxicogenomics in Toxicologic Pathology
Published in Pritam S. Sahota, James A. Popp, Jerry F. Hardisty, Chirukandath Gopinath, Page R. Bouchard, Toxicologic Pathology, 2018
Arun R. Pandiri, David E. Malarkey, Mark J. Hoenerhoff
In accordance with the 3Rs principle, in vitro studies using cell cultures are being used for screening and to decipher the mechanisms of action of hepatic toxicity. Hepatocytic cell cultures with varying degrees of metabolic activity (HepaRG>HepG2) either in 2D or 3D (organoid) configurations are increasingly being used to identify early signals of hepatic toxicity in a high-throughput manner. These signals could be based on traditional toxicity endpoints (cytotoxicity) or toxicogenomic endpoints such as certain mRNA (signatures) and miRNA (miR-122). In addition, toxicogenomic platforms that focus only on a subset of the transcriptome that respond to xenobiotic exposure have also been developed for humans, rodents and zebrafish. One example is the S1550+ TempOSeq platform that is currently being used in the TOX21-Phase III to enable high throughput screening of chemicals in 3D cell cultures and determine the point of departure using the BMD approach (NTP 2017).
Aspergillus
Published in Dongyou Liu, Laboratory Models for Foodborne Infections, 2017
László Kredics, János Varga, Rajagopalaboopathi Jayasudha, Sándor Kocsubé, Nikolett Baranyi, Coimbatore Subramanian Shobana, Muthusamy Chandrasekaran, Shine Kadaikunnan, Venkatapathy Narendran, Csaba Vágvölgyi, Palanisamy Manikandan
Table 31.3 summarizes the studies published since 2010 that have used human or animal cell lines as laboratory models for the investigation of Aspergillus mycotoxins [184–213]. Among the retrieved records, relevant data were found in 31 publications. Eleven studies applied different liver cell lines (human hepatocellular carcinoma cell lines Huh7, HepG2, Hep3B, or SMMC-7721, HepaRG terminally differentiated hepatic cells derived from a human hepatic progenitor cell line that retains many characteristics of primary human hepatocytes, the HeLa derivative Chang liver cell line, as well as H4IIE rat hepatoma cells) for AFB1 research. The major outputs of these studies included the development of a stable Huh7 cell line with constitutive expression of human CYP4501A2 [184], the demonstration that AFB1 is able to activate the pregnane X receptor (PXR), a known regulator of liver xenobiotic metabolism in human hepatocytes [185], the demonstration that AFB1 stimulates hepatoma cell migration [186], the recognition that AFB1 exposure decreases the replication of the hepatitis B virus [190], and the detection of an interaction between AFB1 and FB1 on activation and expression of CYP1A and its transcription factor Ahr [194]. Another group of studies applied various kidney cell lines (RPTEC/TERT1 immortalized human renal proximal tubular epithelial cells, IHKE immortalized human kidney cells, Vero African green monkey kidney cells, or the porcine kidney cell lines LLC-PK1 and PK-15) to study Aspergillus mycotoxins, primarily OTA. Among others, these studies revealed that OTA causes significant deciliation in a model of the proximal tubule [196], that the hypoxia inducible factor 2α (but not 1α) plays a crucial role in prevention of diminishment of vascular endothelial growth factor production evoked by OTA in kidney proximal tubular epithelial cells [197], and that with its high antioxidant potential, quercetin protects Vero cells from OTA-induced oxidative stress and apoptosis [200]. The immunomodulatory or immunotoxic properties of Aspergillus mycotoxins were studied on J774A.1 murine macrophages [201] and human T-cell lines [202,203]. Studies on human colon carcinoma cell lines (Caco-2, HCT116, and SW620) revealed important data about the toxicity of AFB1, OTA, and PAT [204–206]; for the latter, a new anticancer mechanism was proposed [205]. Further cell lines used since 2010 to study Aspergillus mycotoxins include Sp2/0 mouse myeloma cells and 2C10 and 2E6 mouse hybridoma cells [207]; SAOS-2 human osteosarcoma cells [208], MCF-7aro human breast cancer cells, JEG-3 human placental cells, MCF-7 human breast adenocarcinoma cells, and T98G human Caucasian glioblastoma cells [209]; B-13 and B-13/H rat pancreatic progenitor cells [210]; A549 human lung carcinoma cells [211]; INS-1 rat insulinoma cells [212]; as well as L929 murine subcutaneous connective tissue cells and M12.C3.F6 murine B cell lymphoma cells [213].
Comparative cytotoxicity induced by parabens and their halogenated byproducts in human and fish cell lines
Published in Drug and Chemical Toxicology, 2023
Ashley L. Ball, Megan E. Solan, Marco E. Franco, Ramon Lavado
Undifferentiated HepaRG cells were obtained from BioPredic International (Paris, France) (distributors in the USA: Lonza Walkersville Inc., Walkersville, MD) and grown as recommended by Tascher et al. (2019). Briefly, Gibco-formulated William’s E Medium was supplemented with 2 mM Glutamax, 100 U/mL penicillin, 100 µg/mL streptomycin, 10% FBS, 5 µg/mL insulin, and 50 µM hydrocortisone hemisuccinate to constitute HepaRG growth media. Cells were grown at 37 °C in a 5% CO2/air atmosphere. Undifferentiated HepaRG cells were maintained in this growth media for 28 days before undergoing differentiation. When confluence was reached, HepaRG cells were shifted to the same medium supplemented with 2% of DMSO, hereafter referred to as the differentiation medium, for two additional weeks, leading to confluent differentiated cultures containing equal proportions of hepatocyte-like and progenitors/primitive biliary-like cells.
Through a glass, darkly? HepaRG and HepG2 cells as models of human phase I drug metabolism
Published in Drug Metabolism Reviews, 2022
Lesley A. Stanley, C. Roland Wolf
The patent-protected HepaRG cell line (PCT/FR02/02391, 2002) was established from a Grade I hepatocarcinoma growing in an adult female with chronic hepatitis C infection. The resulting cultures comprised bipolar oval cells with the capacity to differentiate when treated with dimethyl sulfoxide (DMSO; ∼2% in cell culture medium) and hydrocortisone (50 µM) (Parent et al. 2004). The resulting hepatocyte-like cells metabolized phenacetin, tolbutamide, dextromethorphan, and nifedipine with specific activities ∼2–10 pmol/min/mg protein (Gripon et al. 2002), while fluorescence-based assays demonstrated 7-ethoxyresorufin O-deethylation (EROD), tolbutamide 4-hydroxylation, testosterone 6β-hydroxylation and chlorzoxazone 6-hydroxyation in differentiated HepaRGs; EROD activity was upregulated by 3-MC (5 µM, 24 h) but testosterone 6β-hydroxylation was not further increased in response to the CYP3A4 inducer rifampicin (25 µM, 72 h) (Aninat et al. 2006).
The development and hepatotoxicity of acetaminophen: reviewing over a century of progress
Published in Drug Metabolism Reviews, 2020
Mitchell R. McGill, Jack A. Hinson
Additional translational data have come from recent studies using cultured human hepatocytes. HepaRG is a unique bipotential human liver cell line that can differentiate into hepatocytes and bile duct epithelium-like cells. Importantly, HepaRG cells express most major drug metabolizing enzymes and transporters at levels similar to PHHs. It has been demonstrated that APAP is toxic specifically to hepatocytes within HepaRG cultures, with a time course of injury that resembles APAP overdose patients (McGill et al. 2011). APAP also causes GSH depletion, protein binding, mitochondrial depolarization and oxidative stress in HepaRG cells, and each of those precedes the cell death, indicating that they may be important for the mechanism (McGill et al. 2011). More recently, the mechanisms of APAP toxicity were explored in PHHs freshly isolated from resected liver tissue. Like HepaRG cells, fresh PHH develop toxicity with a time course that resembles humans (Xie et al. 2014). And, again, mitochondrial depolarization and oxidative stress precede the cell death. Importantly, JNK activation was also observed in PHH after APAP treatment, and the JNK inhibitor SP600125 reduced APAP toxicity in that model (Xie et al. 2014). Finally, there is evidence that 4-methylpyrazole (4MP) protects PHH against APAP toxicity by inhibiting cytochromes P450 and reducing protein alkylation (Akakpo et al. 2018). Collectively, the data from APAP overdose patients and cultured human hepatocytes strongly indicate that the basic mechanisms of APAP hepatotoxicity in humans and mice are at least similar, if not identical.