Trehalose is a natural disaccharide formed by a 1,1-glycosidic bond between two α-glucose units, widely used in food industry due to its peculiar chemical properties. In the last 20 years the involuntary trehalose intake is exponentially increased, with unknown long-term effects on the population. Furthermore, circulating trehalose is able to reach the CNS. Importantly, trehalose induces autophagy in an mTORC1-independent manner activating the transcription factor EB (TFEB), the master transcriptional regulator of autophagy-lysosomal biogenesis. Accordingly, several studies indicated that trehalose promotes the degradation of accumulated proteins and organelles with beneficial neuroprotective effects in neurodegenerative models, both in-vitro and in-vivo, with elusive molecular mechanisms.In our laboratory, using biochemistry, liquid chromatography coupled with mass spectrometry (LC-MS) and patch-clamp techniques, we aim to investigate: 1) the molecular pathways underlying trehalose-dependent autophagy activation; 2) the effect of trehalose on neuronal functions. Our preliminary results indicate that 25 mM trehalose in-vitro treatment increases the expression of autophagic markers (p62 and LC3-II), indicating an activation of the autophagy cascade, as expected. In parallel, LC-MS analyses indicate an altered energetic and aminoacidic metabolism, occurring after 1h of trehalose treatment, suggesting the metabolic change as the triggering factor for autophagy induction. Intriguingly, after 1h treatment we found significant trehalose accumulation in LAMP1 positive isolated lysosomes.Moreover, we found that animals chronically treated with trehalose present altered neuronal excitability, highlighting also gender differences. In conclusion, our preliminary results indicate that trehalose accumulates in lysosomes and it is able to modify neuronal metabolism and functions.