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Application of Cell Painting, an Imaging-Based High Throughput Phenotypic Profiling Assay for Bioactivity Screening of Environmental Chemicals (SOT 2021)
Citation:
Nyffeler, J. AND J. Harrill. Application of Cell Painting, an Imaging-Based High Throughput Phenotypic Profiling Assay for Bioactivity Screening of Environmental Chemicals (SOT 2021). Society of Toxicology Virtual Annual Meeting 2021, virtual, NC, March 15 - 25, 2021. https://doi.org/10.23645/epacomptox.16799392
Impact/Purpose:
Presentation to the Society of Toxicology (SOT) virtual annual meeting in March 2021.
Description:
Cell Painting is an imaging-based high throughput phenotypic profiling (HTPP) assay that uses fluorescent probes to label a variety of organelles (e.g., nucleus, endoplasmic reticulum, Golgi, mitochondria) and measures a large number of features at the cellular level (e.g., intensity, texture, localization of signal) to detect chemical-induced changes in cell morphology. This assay can be deployed in high throughput screening format across multiple human-derived in vitro models and generate high content data (n = 1300 measured features/cell) that can be leveraged to identify potency thresholds for perturbation of cellular biology and inform putative mechanism-of-action prediction for next-generation risk assessment ). Here, we describe HTPP screening assay design, workflows for phenotypic feature extraction, open-source approaches for concentration-response modeling of high-dimensional data, evaluation of assay reproducibility using phenotypic reference chemicals, and results from a concentration-response screening of >1,200 chemicals from the ToxCast chemical library in U-2 OS cells treated for 24 hours. Overall, 41% of the chemicals produced a change in cell phenotype and were classified as hits in the HTPP assay. Where possible, based on the availability of high throughput toxicokinetic modeling data, micromolar potency values from active chemicals in the HTPP assay were converted to administered equivalent doses (AEDs) using in vitro to in vivo extrapolation and reverse dosimetry. For many chemicals, AEDs based on HTPP bioactivity potencies were higher than predicted human exposures and lower or comparable to in vivo effect values from mammalian toxicity studies. Using profile comparison methods, we observed that profiles for retinoic acid receptor agonists (e.g., arotinoid acid, bexarotene) and glucocorticoid receptor agonists (e.g., betamethasone, budesonide) were similar to their respective model compounds (retinoic acid, dexamethasone). In addition, profile similarities were observed for several different classes of pesticides (e.g., organochlorines, strobilurins). Participants in this session will gain a broader understanding of imaging-based high throughput profiling methods and potential applications relating to chemical grouping and prioritization in the context of chemical safety assessment. This abstract does not reflect US EPA policy.
URLs/Downloads:
DOI: Application of Cell Painting, an Imaging-Based High Throughput Phenotypic Profiling Assay for Bioactivity Screening of Environmental Chemicals (SOT 2021)
SOT2021_NYFFELER V3.PDF (PDF, NA pp, 7235.057 KB, about PDF)