Related Experiment Video
Updated: May 13, 2026
An Intestine/Liver Microphysiological System for Drug Pharmacokinetic and Toxicological Assessment
Published on: December 3, 2020
Comparative case study on NAMs: towards enhancing specific target organ toxicity analysis
Kristina Jochum1, Andrea Miccoli1,2,3, Cornelia Sommersdorf4
1Department of Pesticides Safety, German Federal Institute for Risk Assessment, Berlin, Germany.
Abstract:
Traditional risk assessment methodologies in toxicology have relied upon animal testing, despite concerns regarding interspecies consistency, reproducibility, costs, and ethics. New Approach Methodologies (NAMs), including cell culture and multi-level omics analyses, hold promise by providing mechanistic information rather than assessing organ pathology. However, NAMs face limitations, like lacking a whole organism and restricted toxicokinetic interactions. This is an inherent challenge when it comes to the use of omics data from in vitro studies for the prediction of organ toxicity in vivo. One solution in this context are comparative in vitro-in vivo studies as they allow for a more detailed assessment of the transferability of the respective NAM data. Hence, hepatotoxic and nephrotoxic pesticide active substances were tested in human cell lines and the results subsequently related to the biology underlying established effects in vivo. To this end, substances were tested in HepaRG and RPTEC/tERT1 cells at non-cytotoxic concentrations and analyzed for effects on the transcriptome and parts of the proteome using quantitative real-time PCR arrays and multiplexed microsphere-based sandwich immunoassays, respectively. Transcriptomics data were analyzed using three bioinformatics tools. Where possible, in vitro endpoints were connected to in vivo observations. Targeted protein analysis revealed various affected pathways, with generally fewer effects present in RPTEC/tERT1. The strongest transcriptional impact was observed for Chlorotoluron in HepaRG cells (increased CYP1A1 and CYP1A2 expression). A comprehensive comparison of early cellular responses with data from in vivo studies revealed that transcriptomics outperformed targeted protein analysis, correctly predicting up to 50% of in vivo effects.
Insights
New Approach Methodologies (NAMs) show promise for toxicology, but linking in vitro omics data to in vivo organ toxicity remains challenging. Transcriptomics analysis of cell lines accurately predicted up to 50% of in vivo effects, outperforming protein analysis.
Area of Science:
- Toxicology and Ecotoxicology
- In Vitro Toxicology
- Omics Technologies
Background:
- Traditional toxicology relies on animal testing, facing ethical and reproducibility concerns.
- New Approach Methodologies (NAMs) offer mechanistic insights but struggle with whole-organism and toxicokinetic limitations.
- Bridging in vitro omics data to in vivo organ toxicity prediction is a significant challenge.
Purpose of the Study:
- To evaluate the transferability of NAM data by conducting comparative in vitro-in vivo studies.
- To assess the predictive power of transcriptomics and targeted proteomics from human cell lines for organ toxicity.
- To connect early cellular responses in vitro to established in vivo effects of pesticide active substances.
Main Methods:
- HepaRG and RPTEC/tERT1 human cell lines were exposed to hepatotoxic and nephrotoxic pesticide active substances at non-cytotoxic concentrations.
- Transcriptome-wide gene expression was analyzed using quantitative real-time PCR arrays.
- Proteome analysis was performed using multiplexed microsphere-based sandwich immunoassays.
- Bioinformatics tools were employed to analyze transcriptomics data and link in vitro endpoints to in vivo observations.
Main Results:
- Transcriptomics analysis revealed significant cellular responses, with Chlorotoluron showing the strongest impact in HepaRG cells (increased CYP1A1 and CYP1A2 expression).
- Targeted protein analysis identified affected pathways, with fewer effects observed in RPTEC/tERT1 cells compared to HepaRG cells.
- Transcriptomics data demonstrated a higher predictive capacity, correctly identifying up to 50% of observed in vivo effects, outperforming targeted protein analysis.
Conclusions:
- Comparative in vitro-in vivo studies are valuable for assessing NAM data transferability.
- Transcriptomics analysis of human cell lines shows significant potential for predicting in vivo organ toxicity.
- NAMs, particularly transcriptomics, offer a promising alternative to traditional animal testing in toxicology risk assessment.
