Pancreatic cell derived extracellular vesicles containing surface preproinsulin are involved in glucose stimulated insulin secretion

Aims: Extracellular vesicles (EVs) are involved in intercellular communication and are a topic of increasing interest due to their therapeutic potential. The aim of this study was to determine whether human islet -derived EVs contain insulin, and if so, what role do they play in glucose stimulated insulin secretion. Main methods: We isolated EVs from human islets culture and plasma to probe for insulin. Plasma from hyperglycemic glucose clamp experiments were also used to isolate and measure EV insulin content in response to a secretory stimulus. We performed immunogold electron microscopy for insulin presence in EVs. Co -culture experiments of isolated EVs with fresh islets were performed to examine the effect of EV cargo on insulin receptor signaling. Key findings: EVs isolated from culture medium contained insulin. Glucose treatment of islets increased the level of EV insulin. Hyperglycemic glucose clamp experiments in humans also lead to increased levels of insulin in plasma -derived EVs. Immunogold electron microscopy and proteinase K -digestion experiments demonstrated that insulin in EVs predominantly associated with the exterior surface of EVs while western blot analyses uncovered the presence of only preproinsulin in EVs. Membrane -bound preproinsulin in EVs was capable of activating insulin signaling pathway in an insulin receptor -dependent manner. The physiological relevance of this finding was observed in priming of human naive islets by EVs during glucose stimulated insulin secretion. Significance: Our data suggest that (1) human islets secret insulin via an alternate pathway (EV-mediated) other than conventional granule -mediated insulin secretion, and (2) EV membrane bound preproinsulin is biologically active.