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Updated: Dec 31, 2025
A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans
Published on: March 14, 2019
Bioactivity screening of environmental chemicals using imaging-based high-throughput phenotypic profiling
Johanna Nyffeler1, Clinton Willis2, Ryan Lougee1
1Center for Computational Toxicology and Exposure, Office of Research and Development, US Environmental Protection Agency, Durham, NC 27711, United States of America; Oak Ridge Institute for Science and Education (ORISE), Oak Ridge, TN 37831, United States of America.
This study adapted the Cell Painting assay for screening environmental chemical bioactivity. The high-throughput phenotypic profiling (HTPP) assay efficiently identifies chemical effects on cell morphology, aiding in safety assessments.
Area of Science:
- Toxicology
- Cell Biology
- High-Throughput Screening
Background:
- Environmental chemical safety assessments require robust screening methods.
- Existing methods may not fully capture chemical bioactivity.
- High-throughput phenotypic profiling (HTPP) offers a potential solution.
Purpose of the Study:
- Adapt the Cell Painting assay for HTPP of environmental chemicals.
- Develop a workflow for analyzing phenotypic data and determining bioactivity thresholds.
- Compare in vitro thresholds with in vivo toxicity data.
Main Methods:
- Adapted Cell Painting assay using U-2 OS cells and fluorescent probes.
- Screened 14 reference chemicals and 462 ToxCast library chemicals in concentration-response.
- Developed novel workflow for feature extraction, modeling, and threshold determination.
- Utilized reverse dosimetry to calculate administered equivalent doses (AEDs).
Main Results:
- Validated reference chemicals produced expected phenotypic changes.
- Developed and implemented a novel workflow for data analysis and threshold determination.
- HTPP-derived AEDs were comparable or more conservative than in vivo effect values in 68% of cases.
Conclusions:
- The adapted HTPP assay is an efficient, cost-effective, and reproducible method for screening environmental chemicals.
- This approach aids in characterizing biological activity and potency for in vitro safety assessments.
- HTPP provides valuable data for understanding chemical impacts on cellular morphology and potential toxicity.
