Morphological profiling of environmental chemicals enables efficient and untargeted exploration of combination effects

Abstract

Environmental chemicals are commonly studied one at a time, and there is a need to advance our understanding of the effect of exposure to their combinations. Here we apply high-content microscopy imaging of cells stained with multiplexed dyes (Cell Painting) to profile the effects of Cetyltrimethylammonium bromide (CTAB), Bisphenol A (BPA), and Dibutyltin dilaurate (DBTDL) exposure on four human cell lines; both individually and in all combinations. We show that morphological features can be used with multivariate data analysis to discern between exposures from individual compounds, concentrations, and combinations. CTAB and DBTDL induced concentration-dependent morphological changes across the four cell lines, and BPA exacerbated morphological effects when combined with CTAB and DBTDL. Combined exposure to CTAB and BPA induced changes on the ER, Golgi apparatus, nucleoli and cytoplasmic RNA in one of the cell lines. Different responses between cell lines indicate that multiple cell types are needed when assessing combination effects. The rapid and relatively low-cost experiments combined with high information content makes Cell Painting an attractive methodology for future studies of combination effects. All data in the study is made publicly available on Figshare. HighlightsO_LIAssessment of combination effects of BPA, CTAB and DBTDL on four human cell lines C_LIO_LIMorphological profiling/Cell Painting captures dose and combination dependent effects C_LIO_LIBPA exacerbated morphological effects when combined with CTAB and DBTDL. C_LIO_LICell models of diverse origin are needed when profiling environmental chemicals C_LI