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Identifying putative mode-of-actions for environmental chemicals using high-throughput phenotypic profiling (WC11)
Citation:
Nyffeler, J., C. Willis, G. Patlewicz, D. Chang, I. Shah, AND J. Harrill. Identifying putative mode-of-actions for environmental chemicals using high-throughput phenotypic profiling (WC11). 11th World Conference on Alternatives and Animal Use in the Life Science (WC11), Virtual, NC, NETHERLANDS, August 23 - September 02, 2021. https://doi.org/10.23645/epacomptox.16837921
Impact/Purpose:
Poster submitted to the 11th World Conference on Alternatives and Animal Use in the Life Science in August 2021 in Maastrich. The Environmental Protection Agency is exploring the use of profiling methods for rapid bioactivity screening and hazard evaluation of environmental chemicals. Phenotypic profiling is an imaging-based method that measures morphological features of organelles at the single cell level.
Description:
The United States Environmental Protection Agency (USEPA) is exploring the use of high-throughput profiling methods for rapid bioactivity screening of environmental chemicals. In phenotypic profiling using the Cell Painting assay, cell organelles are labeled with multiple fluorophores to quantify morphological changes of cells upon chemical exposure. While it is well known that drug-like chemicals can be grouped by mode-of-action (MOA) based on the phenotypic profiles, here we explore whether this is also true for environmental chemicals (i.e. pesticides, industrial chemicals, etc.) that might lack a primary target in human cells. To this end, we screened 120 chemicals with annotated MOAs (‘reference chemicals’) and 441 environmental chemicals at 8 concentrations (1/2 log10 spacing, 4 biological replicates) in U-2 OS cells. Cells were exposed for 24 h, fixed and labeled with fluorescent dyes to visualize multiple organelles: nucleus, nucleoli, endoplasmic reticulum, golgi, actin cytoskeleton, plasma membrane and mitochondria. 1300 features were extracted per cell to construct profiles, which were then compared to each other using Pearson correlation. Among the reference chemicals, multiple distinct profiles were observed. For example, various DNA damaging compounds (alkylators, topoisomerase inhibitors, antimetabolites) have similar profiles, while microtubule modulators had a different, characteristic profile. Among all tested chemicals, approximately 16 clusters were observed, and 141/441 environmental chemicals clustered with reference chemicals, indicating that they produce similar phenotypes and hence might have similar cellular effects. A group of strobilurin fungicides had a common profile related to changes in mitochondrial features, consistent with strobilurin’s MOA to inhibit mitochondrial respiration in fungi. Overall these preliminary results indicate (1) that our screening strategy yields distinct phenotypic profiles and (2) that environmental chemicals may be grouped using these profiles to identify putative MOAs. This abstract does not reflect USEPA policy.
URLs/Downloads:
DOI: Identifying putative mode-of-actions for environmental chemicals using high-throughput phenotypic profiling (WC11)
WC11_NYFFELER_POSTER_V3_CLEARED_M.PDF (PDF, NA pp, 3107.525 KB, about PDF)