Experimental evidence of a limited impact of new-generation perfluoroalkyl substance C6O4 on differentiating human dopaminergic neurons from induced pluripotent stem cells

• Published in: Toxicology reports Vol. 10; pp. 40 - 44
• Main Authors: Di Nisio, Andrea, Trevisan, Marta, Dall’Acqua, Stefano, Pannella, Micaela, Pappalardo, Claudia, Ferlin, Alberto, Foresta, Carlo, De Toni, Luca
• Format: Journal Article
• Language: English
• Published: Ireland Elsevier B.V 01-01-2023; Elsevier
• Subjects: Dopaminergic neuron; Endocrine disruptors; Human induced pluripotent stem cells; Perfluoroalkyl substances; Tyrosine hydroxylase; Endocrine disruptors; Human induced pluripotent stem cells; Dopaminergic neuron; Tyrosine hydroxylase; Perfluoroalkyl substances
• ISSN: 2214-7500; 2214-7500
• DOI: 10.1016/j.toxrep.2022.12.006

Perfluoroalkyl substances (PFASs) are persistent pollutants, raising concerns for human health. Legacy PFAS perfluoro-octanoic acid (PFOA) accumulate in brains of people at high environmental exposure, especially in areas enriched with dopaminergic neurons (DN). In vitro exposure to 10 ng/mL PFOA for 24 h was also associated with an altered molecular and functional phenotype of DN differentiated from human induced pluripotent stem cells (hiPSCs). Acetic acid, 2,2-difluoro-2-((2,2,4,5-tetrafluoro-5(trifluoromethoxy)− 1,3-dioxolan-4-yl)oxy)-ammonium salt (1:1), known as C6O4, is a new generation PFAS proposed to have a safer profile. Here we investigated the effect of C6O4 exposure on the molecular phenotype of hiPSC-derived DN. Cells were exposed to C6O4 for 24 h, at the concentration of 10 ng/mL, at neuronal commitment (DP1), neuronal precursor (DP2) and the mature dopaminergic (DP3) phases of differentiation. Liquid-chromatography/mass-spectrometry showed negligible cell accumulation of C6O4 at each differentiation stage and by staining with Merocyanine-540 we observed unaltered cell membrane fluidity. Immunofluorescence showed that the expression of tyrosine hydroxylase (TH) and βIII-Tubulin was unaffected by the exposure to C6O4 at each differentiation phase (respectively: DP1, p = 0.332; DP2, p = 0.623; DP3, p = 0.816, with respect to control unexposed conditions). Exposure to C6O4 is presumed to have minor effects on cell molecular/functional phenotype of developing human DN cells, requiring confirm on in vivo models.
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•Legacy perfloroalkyl substances (PFAS) interfere with dopaminergic neurons differentiation.•Little is known about new generation PFAS, like C6O4, effect on neurons.•We investigated the effect of C6O4 on the molecular phenotype of dopaminergic neurons.•C6O4 has minor effects on cell phenotype of developing human dopaminergic neurons.