A Shotgun\nProteomic-Based\nApproach with a Q‑Exactive\nHybrid Quadrupole-Orbitrap High-Resolution Mass Spectrometer for the\nAssessment of Pesticide Mixture-Induced Neurotoxicity on a 3D-Developed\nNeurospheroid Model from Human Brain Meningiomas: I

The widespread use\nof pesticides, particularly in combinations,\nhas resulted in enhanced hazardous health effects. However, little\nis known about their molecular mechanism of interactions. The aim\nof this study was to assess the neurotoxicity effect of pesticides\nin mixtures by adopting a 3D in vitro developed neurospheroid model,\nfollowed by treatment by increased concentrations of pesticides for\n24 h and analysis by a shotgun proteomic-based approach with high-resolution\ntandem mass spectrometry. Three proteins, namely, glyceraldehyde-3-phosphate-dehydrogenase\n(GAPDH), α-enolase, and phosphoglycerate-kinase-1, were selected\nas key targets in the metabolic process. Only high doses of pesticides\nmitigated cell-density proliferation with the occurrence of apoptotic\ncells, which unlikely makes any neurological alterations in environmental\nregulatory exposures. The proteomic analysis showed that majority\nof altered proteins were implicated in cell metabolism. De novo peptide\nsequencing revealed ion losses and adduct formation, namely, a trityl-post-translational\nmodification in the active site of 201-GAPDH protein. The study also\nhighlights the plausible role of pyrethroids to be implicated in the\ndeleterious effects of pesticides in a mixture. To the best of our\nknowledge, our finding is the first in toxicoproteomics to deeply\nelucidate pesticides’ molecular interactions and their ability\nto adduct proteins as a pivotal role in the neurotoxicity mechanism.