An Automated Multiplexed Hepatotoxicity and CYP Induction Assay Using HepaRG Cells in 2D and 3D

Ott, Lindsey; Ramachandran, Karthik; Stehno‐Bittel, Lisa

doi:10.1177/2472555217701058

Cited by 40 publications

(45 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…HepaRG spheroids can be cultured up to 7 weeks with polarized transporters and bile canalicular structures (Leite et al, 2012). Furthermore, activities of CYP1A2, CYP2B6, CYP2C9, CYP3A4, and UGT enzymes remained stable and substantially (2- to 20-fold) higher in 3D compared to 2D culture (Leite et al, 2012; Wang et al, 2015; Ott et al, 2017). In accordance with these functional data spheroid culture improved sensitivity to acetaminophen and aflatoxin B1, which both require metabolic activation (Leite et al, 2012; Gunness et al, 2013; Mueller et al, 2014).…”

Section: Organotypic Liver Models For Hepatotoxicity Predictionsmentioning

confidence: 94%

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

2019

View full text Add to dashboard Cite

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.

show abstract

Section: Organotypic Liver Models For Hepatotoxicity Predictionsmentioning

confidence: 94%

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

2019

View full text Add to dashboard Cite

show abstract

“…The same holds for HepaRG cells that still show slightly more non-DEGs than HepG2 cells but are less similar to in vivo liver tissue as PHH or microtissues. Several recent studies reported that HepaRG 3D models mimic in vivo liver (Ott et al 2017;Ramaiahgari et al 2017;Takahashi et al 2015) but these studies did not perform an RNA-Seq based comparison to human liver tissues.…”

Section: Discussionmentioning

confidence: 99%

Comparing in vitro human liver models to in vivo human liver using RNA-Seq

et al. 2020

View full text Add to dashboard Cite

The liver plays an important role in xenobiotic metabolism and represents a primary target for toxic substances. Many different in vitro cell models have been developed in the past decades. In this study, we used RNA-sequencing (RNA-Seq) to analyze the following human in vitro liver cell models in comparison to human liver tissue: cancer-derived cell lines (HepG2, HepaRG 3D), induced pluripotent stem cell-derived hepatocyte-like cells (iPSC-HLCs), cancerous human liver-derived assays (hPCLiS, human precision cut liver slices), non-cancerous human liver-derived assays (PHH, primary human hepatocytes) and 3D liver microtissues. First, using CellNet, we analyzed whether these liver in vitro cell models were indeed classified as liver, based on their baseline expression profile and gene regulatory networks (GRN). More comprehensive analyses using non-differentially expressed genes (non-DEGs) and differential transcript usage (DTU) were applied to assess the coverage for important liver pathways. Through different analyses, we noticed that 3D liver microtissues exhibited a high similarity with in vivo liver, in terms of CellNet (C/T score: 0.98), non-DEGs (10,363) and pathway coverage (highest for 19 out of 20 liver specific pathways shown) at the beginning of the incubation period (0 h) followed by a decrease during long-term incubation for 168 and 336 h. PHH also showed a high degree of similarity with human liver tissue and allowed stable conditions for a short-term cultivation period of 24 h. Using the same metrics, HepG2 cells illustrated the lowest similarity (C/T: 0.51, non-DEGs: 5623, and pathways coverage: least for 7 out of 20) with human liver tissue. The HepG2 are widely used in hepatotoxicity studies, however, due to their lower similarity, they should be used with caution. HepaRG models, iPSC-HLCs, and hPCLiS ranged clearly behind microtissues and PHH but showed higher similarity to human liver tissue than HepG2 cells. In conclusion, this study offers a resource of RNA-Seq data of several biological replicates of human liver cell models in vitro compared to human liver tissue.

show abstract

“…It is important to note that differences in cell viability have been rather inconsistent in the literature. Studies comparing cell viability in hepatocyte monolayer and spheroid cultures found that spheroids required a higher (Walker et al 2000) or lower (Ott et al 2017) concentration to induce cytotoxicity compared with confluent cells. A chemotherapy drug had similar cytotoxicity in confluent and spheroid cultures of the prostate adenocarcinoma cell line LNCaP, whereas monolayer cells of the same cell line were more sensitive to another anticancer drug (Takagi et al 2007).…”

Section: Pcb-126-induced Cytotoxicitymentioning

confidence: 99%

Evaluation of the Aryl Hydrocarbon Receptor Response in LMH 3D Spheroids

Sharin

Crump

O’Brien

2020

Enviro Toxic and Chemistry

View full text Add to dashboard Cite

In the present study, we investigated whether the immortalized chicken hepatocellular carcinoma cell line, leghorn male hepatoma (LMH), had a comparable aryl hydrocarbon receptor (AhR) response to primary chicken embryonic hepatocytes (CEHs) when used in a well‐established assay for chemical screening and prioritization. The LMH cells were grown as 2‐dimensional (2D) confluent cells and 3D spheroids to determine the optimal cell culture states for chemical screening. Cytochrome P450 1A4 and 1A5 (CYP1A) activity and gene expression were compared between CEHs and LMH cells grown in 2 culture states following exposure to the dioxin‐like compound 3,3′,4,4′,5‐pentachlorobiphenyl (PCB‐126). The CYP1A activity was measured using the ethoxyresorufin‐O‐deethylase (EROD) assay, and changes in mRNA expression associated with the AhR pathway were determined using a custom‐designed polymerase chain reaction array. Among LMH cell culture states (i.e., 2D vs 3D), EROD induction was observed only in 3D LMH spheroids. Similarly, 3D spheroids had the greatest number of changes in AhR‐related genes compared with confluent cells. Overall, these results suggest that LMH cells grown as 3D spheroids have a metabolic and gene expression profile that is comparable to that of CEH, and may represent a suitable animal‐free alternative for in vitro screening of chemicals. Environ Toxicol Chem 2020;39:1693–1701. © 2020 SETAC

show abstract

An Automated Multiplexed Hepatotoxicity and CYP Induction Assay Using HepaRG Cells in 2D and 3D

Cited by 40 publications

References 47 publications

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

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

Comparing in vitro human liver models to in vivo human liver using RNA-Seq

Evaluation of the Aryl Hydrocarbon Receptor Response in LMH 3D Spheroids

Contact Info

Product

Resources

About