Currently, it is an open question how gut senses metabolic alterations and whether this is transmitted to other tissues via EVs. Here, we observed that protein packaging in gut derived EVs (GDE) from a diet induced-prediabetic mouse mode, specifically small intestine, is increased in prediabetes. Proteins related to lipid metabolism and to oxidative stress management were more abundant in prediabetic GDE compared to healthy controls. Low-fat diet: A1 to A3 high-fat diet: A4 to A6