Calibrated flow cytometry enables rapid detection of CD45−EpCAM+ extracellular vesicles for pancreatic cancer clinical assessment
Reliable non-invasive biomarkers are needed to optimize treatment selection and enable real-time monitoring of pancreatic cancer (PC). We developed a bead-calibrated, multiparametric flow cytometry (FC) protocol to detect, enumerate, and subtype circulating intact extracellular vesicles (EVs) in blood. Among 120 individuals analyzed by this platform, patients with PC had significantly higher levels of CD45 − EpCAM + EVs than controls. These EVs correlated positively with advanced tumor stage, while their combination with serum CA19-9 improved monitoring accuracy in an exploratory cohort. Findings were supported by two PC mouse models, in which circulating CD45 − EpCAM + EVs correlated with tumor volume. Remarkably, high baseline levels were associated with chemotherapy non-response and shorter overall survival. Overall, this practical, bead-calibrated FC method identified circulating CD45 − EpCAM + EVs as a promising biomarker candidate for PC. The platform may facilitate robust detection of new circulating EV markers and support disease monitoring and treatment stratification.
Authors
- Rossella Crescitelli (ORCID: https://orcid.org/0000-0002-1714-3169)
- Fabio F. di Mola (ORCID: https://orcid.org/0000-0001-9877-2946)
- Paola Lanuti (ORCID: https://orcid.org/0000-0003-4197-9780)
- Antonino Grassadonia (ORCID: https://orcid.org/0000-0002-2459-0661)
- Anna Piro (ORCID: https://orcid.org/0009-0000-8076-2491)
- Laura De Lellis (ORCID: https://orcid.org/0000-0002-6460-2508)
- Nicola Tinari (ORCID: https://orcid.org/0000-0001-8449-5407)
- Dimitri Luisi
- Pasquale Simeone (ORCID: https://orcid.org/0000-0003-4430-480X)
- Davide Brocco (ORCID: https://orcid.org/0000-0001-8634-7184)
- Pietro Di Marino (ORCID: https://orcid.org/0000-0002-1330-683X)
- Alessandro Cama (ORCID: https://orcid.org/0000-0003-3647-1368)
- Alessia Lamolinara (ORCID: https://orcid.org/0000-0002-7421-4509)
- Mauro Di Ianni (ORCID: https://orcid.org/0000-0002-6701-4023)
- Manuela Iezzi (ORCID: https://orcid.org/0000-0002-6296-6498)
- Rosalba Florio (ORCID: https://orcid.org/0000-0002-3095-1451)
- Serena Veschi (ORCID: https://orcid.org/0000-0003-0450-9210)
- Tommaso Grottola (ORCID: https://orcid.org/0000-0001-5779-3732)
- Serena Pilato (ORCID: https://orcid.org/0000-0003-4971-6483)
- Gianluca Russo (ORCID: https://orcid.org/0000-0003-2698-9470)
- Arianna Aquilini Mummolo
- Francesca D’Ascanio (ORCID: https://orcid.org/0000-0002-0588-5536)
- Marta Caporale
- Giulia Colasante
- Michele De Tursi
- Pierluigi Di Sebastiano
Institutions
- University of Foggia (IT)
- University of Chieti-Pescara (IT)
- Casa di cura Pierangeli (IT)
- Ospedale SS. Annunziata (IT)
- University of Gothenburg (SE)
Publication Details
- Journal
- iScience
- Published
- 2026-10-27
- DOI
- https://doi.org/10.1016/j.isci.2026.118051
- Primary Topic
- Extracellular vesicles in disease
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- European Commission
- Ministero dell’Istruzione, dell’Università e della Ricerca
- Vetenskapsrådet
- Stiftelsen Assar Gabrielssons Fond
- Università degli Studi G. d'Annunzio Chieti - Pescara
- Magnus Bergvalls Stiftelse
- Ministero dell'Istruzione e del Merito