Unlocking the Diagnostic Potential of Extracellular Vesicles in Cancer.

Extracellular vesicles (EVs) are nanoscale lipid bilayer particles that are secreted by virtually all cells into biofluids;their cargo of nucleic acids, proteins, lipids, and glycans is tumor type and status informative and can be measuredvia minimally invasive liquid biopsy. Herein, we evaluate critically the EV-based assay diagnostic accuracy in bothsolid and hematologic malignancies, particularly with regard to exemplar biomarker panels and single-part analyticsfor pancreatic, prostate, breast, lung, and colorectal cancers as well as hematologic disorders. We emphasizenewer immune- and affinity-directed isolation/phenotyping technologies that are more selective and scalable thanconventional ultracentrifugation—e.g., microarray capture (EV Array), EV enrichment from cancer cells, magneticnanopore capture, and filtration/thermophoretic routes—along with single-EV readouts for increased informationalcontent. We also put EV diagnostics in a complementary role to established liquid biopsy modalities (e.g., ctDNA/cfDNA) to maximize detection and disease monitoring, rather than as alternatives. Finally, we outline theranosticapplications, including engineered EVs as delivery vehicles and stromal EVs as therapy resistance biomarkers andmediators, to bridge diagnostics with interventional strategies. This roadmap centers clinically meaningful use casesand methodological stringency to drive translation of EV assays into oncology practice