Abstract 3264: Digital profiling of tumor-derived extracellular vesicle RNA: a novel approach for early diagnosis and monitoring of pancreatic cancer patients

Abstract Introduction: Pancreatic cancer remains a leading cause of cancer-related deaths due to challenges in early diagnosis and the lack of effective biomarkers. Current diagnostic methods are insufficient, necessitating novel approaches. This study explores the use of extracellular vesicle RNA (EV-RNA) from plasma as a diagnostic tool, leveraging a new assay that facilitates membrane fusion between EVs and liposomes. Methods: The study introduces a digital EV-RNA profiling assay (LiquiDyne) that uses charge-mediated fusion between extracellular vesicles and charged liposomes (CLIPs) carrying molecular beacons. This process occurs on a droplet microfluidic chip, where the surface charge of CLIPs is adjusted to optimize fusion efficiency. The assay was validated analytically using EVs from pancreatic cancer cell lines (MIA-PaCa-2, PANC-1, PSN-1, Capan-2) with known KRAS mutations (G12C, G12D, G12R, G12V). Clinical validation involved testing retrospective patient samples across all stages of pancreatic cancer. Results: The assay requires only 20 µL of plasma, removing the need for prior EV isolation or RNA preparation and thus minimizing sample loss. It successfully detected KRAS mutations in both cell lines and patient samples with high accuracy, demonstrating its potential as a reliable diagnostic tool. Conclusions: This innovative assay offers a promising method for early pancreatic cancer diagnosis by enabling precise quantification of rare EV subpopulations. It holds potential not only for diagnosis but also for monitoring minimal residual disease, thus addressing critical gaps in current pancreatic cancer diagnostics. Citation Format: Hyojin Um, Taewon Son, Yeochan Kim, Elizabeth Maria Clarissa, Hanna Kim, Jihye Ahn, Juhee Park, Beomhee Ahn Ahn, Kyusang Lee, Beomseok Lee, Yoon-Kyoung Cho. Digital profiling of tumor-derived extracellular vesicle RNA: a novel approach for early diagnosis and monitoring of pancreatic cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 3264.