Abstract

Humans are exposed to increasing numbers of chemicals that are spreading and accumulating in the environment. Nonetheless, safety assessment and regulations are often still based on toxicity of single chemicals, neglecting combination effects from co-exposure of chemicals. There is a need for new methods to systematically assess combination risks and its implications to human health. Morphological profiling provides an untargeted approach to study cellular responses to various perturbations at low costs, at high throughput and at single cell level. We used the Cell Painting assay to profile the effects of combinations of three environmental chemicals present in everyday objects and contexts. Specifically, we applied multiplexed staining targeting eight relevant cell compartments, followed by high-content microscopy and multiparametric analysis of the cellular profiles. We found that exposure to the chemicals induced distinct morphological responses across four biologically diverse cell lines. Moreover, we demonstrate that the method captures dose and combination-dependent effects. Further work will focus on testing more complex mixtures and on identifying pathways of toxicity of single compounds and combinations.