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Background Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest gastrointestinal cancers with a 5-year survival rate of less than 10%. Biomarkers for early PDAC detection are useful in treating patients with PDAC. Extracellular vesicles (EVs) are lipid-bound vesicles that are potential biomarkers of various diseases such as PDAC. In this study, we quantitatively measured the serum levels of EVs (CD63 + -EVs) or platelet-derived EVs (CD41 + - and CD61 + -EVs) and evaluated their potential use as biomarkers of PDAC. Methods We measured the serum levels of CD63 + -, CD41 + -, CD61 + -EVs using sandwich enzyme-linked immunosorbent assay based on Tim4 with specificity for phosphatidylserine on EVs in age- and sex-matched healthy controls (HCs, n = 39) and patients with PDAC (n = 39). We also examined the effect of tumor burden on the serum EV levels after surgical resection (n = 28). CA19-9, a clinical PDAC biomarker, was also measured for comparison. Results Serum levels of CD63 + -EVs, CD41 + -EVs, and CD61 + -EVs were significantly increased in patients with PDAC compared to HCs. Receiver operating characteristic analysis revealed that CD63 + -EVs exhibited the highest diagnostic performance to discriminate patients with PDAC from HCs (area under the curve (AUC): 0.846), which was comparable to CA19-9 (AUC: 0.842). CA19-9 showed lower AUC values in early stages (I–II, AUC: 0.814) than in late stages (III–IV, AUC: 0.883) PDAC. Conversely, CD63 + -EVs, CD41 + -EVs, and CD61 + -EVs showed comparable AUCs between early- and late-stage PDAC. The combined use of CA19-9 and CD63 + -EVs showed a higher diagnostic performance for early-stage PDAC (AUC: 0.903) than CA19-9. The serum levels of CD63 + -EVs, CD41 + -EVs, CD61 + -EVs, and CA19-9 decreased significantly after surgical resection, demonstrating that EVs are increased in sera of patients depending on the tumor burden. Conclusions The serum levels of CD63 + -EVs and platelet-derived EVs (CD41 + -EVs, CD61 + -EVs) are increased in patients with PDAC than HCs. Since CD63 + -EVs showed a high AUC to discriminate patients with PDAC from HCs; they might be useful as potential biomarkers for PDAC.

CD63-positive extracellular vesicles are potential diagnostic biomarkers of pancreatic ductal adenocarcinoma