Exploring the intracellular incorporation and distribution of PFAS using a novel fluorescent probe

Authors

• EJ Morgan
• A Singh
• Z Shi
• RE Swenson
• CM Guardia

Abstract

Per- and polyfluoroalkyl substances (PFAS), known as “forever chemicals”, pose a serious threat to human health. Their widespread use in commercial and industrial applications has cemented their presence in the environment, causing wildlife and human exposure to them. Due to the chemical stability of PFAS and similar structure to fatty acids, their bioaccumulation has been linked to functional disruptions in a variety of organs. Notably, the placenta is a target tissue for PFAS, resulting in harmful outcomes for both the host and offspring. Using HeLa cells as a model cell line, we explored the incorporation and distribution of PFAS-BODIPY, a novel fluorescent probe, into these cells. Fluorescence-activated cell sorting approaches elucidated the incorporation of PFAS-BODIPY into cells under different culture medium conditions. We found that saturation occurred at much lower concentrations in media lacking fetal bovine serum (FBS), suggesting strong interactions of PFAS with serum components. Additionally, immunofluorescence approaches revealed colocalization of PFAS-BODIPY in the endoplasmic reticulum (ER) and mitochondria, in striking contrast to free BODIPY and lauric acid-BODIPY (RedC12) analogs that accumulate in lipid droplets. These preliminary data suggest a competitive binding model between lipids and PFAS-BODIPY for serum albumin in +FBS media. Furthermore, once inside the cell, PFAS-BODIPY accumulates in different organelles than fatty acids with similar structures. This proof-of-concept compound will provide an innovative visualization tool to study PFAS incorporation and localization in both in vitro and in vivo experiments.

Scientific Focus Area: Cell Biology

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