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Updated: May 3, 2026
High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents HPHC
Published on: May 10, 2016
Quantitative high content imaging of cellular adaptive stress response pathways in toxicity for chemical safety
Steven Wink1, Steven Hiemstra, Suzanna Huppelschoten
1Division of Toxicology, Leiden Academic Centre for Drug Research (LACDR), Leiden University , The Netherlands.
Abstract:
Over the past decade, major leaps forward have been made on the mechanistic understanding and identification of adaptive stress response landscapes underlying toxic insult using transcriptomics approaches. However, for predictive purposes of adverse outcome several major limitations in these approaches exist. First, the limited number of samples that can be analyzed reduces the in depth analysis of concentration-time course relationships for toxic stress responses. Second these transcriptomics analysis have been based on the whole cell population, thereby inevitably preventing single cell analysis. Third, transcriptomics is based on the transcript level, totally ignoring (post)translational regulation. We believe these limitations are circumvented with the application of high content analysis of relevant toxicant-induced adaptive stress signaling pathways using bacterial artificial chromosome (BAC) green fluorescent protein (GFP) reporter cell-based assays. The goal is to establish a platform that incorporates all adaptive stress pathways that are relevant for toxicity, with a focus on drug-induced liver injury. In addition, cellular stress responses typically follow cell perturbations at the subcellular organelle level. Therefore, we complement our reporter line panel with reporters for specific organelle morphometry and function. Here, we review the approaches of high content imaging of cellular adaptive stress responses to chemicals and the application in the mechanistic understanding and prediction of chemical toxicity at a systems toxicology level.
Insights
High content imaging using bacterial artificial chromosome (BAC) green fluorescent protein (GFP) reporter assays overcomes transcriptomics limitations for predicting chemical toxicity. This systems toxicology approach enhances understanding of adaptive stress responses and organelle function.
Area of Science:
- Toxicology and pharmacology
- Cell biology
- Biotechnology
Background:
- Transcriptomics has advanced understanding of adaptive stress responses to toxic insults but has limitations.
- Existing transcriptomics approaches limit in-depth analysis of concentration-time effects, single-cell resolution, and post-translational regulation.
- These limitations hinder accurate prediction of adverse outcomes from chemical exposure.
Purpose of the Study:
- To present high content imaging using bacterial artificial chromosome (BAC) green fluorescent protein (GFP) reporter assays as a method to overcome transcriptomics limitations.
- To establish a platform for analyzing adaptive stress signaling pathways relevant to chemical toxicity, focusing on drug-induced liver injury.
- To integrate organelle morphometry and function reporters to comprehensively assess cellular stress responses.
Main Methods:
- Utilizing high content analysis of BAC-GFP reporter cell-based assays to monitor toxicant-induced adaptive stress signaling pathways.
- Employing reporter lines for specific organelle morphometry and function to complement pathway analysis.
- Applying high content imaging techniques to study cellular adaptive stress responses to chemical compounds.
Main Results:
- High content imaging with BAC-GFP reporters circumvents limitations of traditional transcriptomics for studying chemical toxicity.
- The proposed platform enables detailed analysis of concentration-time dynamics and single-cell responses.
- Integration of organelle-level analysis provides a more holistic view of cellular perturbations.
Conclusions:
- High content imaging of BAC-GFP reporter assays offers a powerful systems toxicology approach for understanding and predicting chemical toxicity.
- This method provides deeper mechanistic insights into adaptive stress responses and organelle integrity.
- The developed platform is crucial for advancing the prediction of adverse outcomes, particularly in drug-induced liver injury.
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