Page 28 - SRPSKO DRUŠTVO ISTRAŽIVAČA RAKA

FUTURE HORIZONS IN CANCER

Manuscript received on: Aug 08, 2025. 616-006-074:577:11
Manuscript revised on: Sep 15, 2025.
Manuscript accepted on: Sep 16, 2025.

Unlocking the diagnostic potential of extracellular vesicles in cancer

Milica Popović
University of Belgrade-Faculty of Chemistry
correspondence: la_bioquimica@chem.bg.ac.rs

Abstract
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 measured
via minimally invasive liquid biopsy. Herein, we evaluate critically the EV-based assay diagnostic accuracy in both
solid and hematologic malignancies, particularly with regard to exemplar biomarker panels and single-part analytics
for pancreatic, prostate, breast, lung, and colorectal cancers as well as hematologic disorders. We emphasize
newer immune- and affinity-directed isolation/phenotyping technologies that are more selective and scalable than
conventional ultracentrifugation—e.g., microarray capture (EV Array), EV enrichment from cancer cells, magnetic
nanopore capture, and filtration/thermophoretic routes—along with single-EV readouts for increased informational
content. 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 theranostic
applications, including engineered EVs as delivery vehicles and stromal EVs as therapy resistance biomarkers and
mediators, to bridge diagnostics with interventional strategies. This roadmap centers clinically meaningful use cases
and methodological stringency to drive translation of EV assays into oncology practice
Keywords: extracellular vesicles, cancer, biomarkers, liquid biopsy.

Introduction
Extracellular vesicles (EVs) are a diverse group of membranous nano-sized structures secreted by cells into the
extracellular space. According to the manner of biogenesis EVs can be classified into exosomes, microvesicles and
apoptotic bodies. (1)Exosomes typically range in size from 30 to 150 nm. They are derived from the endosomal pathway
more specifically by inward budding of the membrane of the early endosomeand formation of intraluminal vesicles
(ILV). The content of ILVs is carefully sorted and packed into multivesicular bodies (MVB)(2). MVB fuse with the plasma
membrane to release exosomes into the extracellular space. Microvesicles are formed by direct outward budding of
plasma membrane. Their typical size ranges from 100 nm to 1 µM. The biogenesis mechanism involves trafficking of
molecular cargo to the plasma membrane, a redistribution of membrane lipids, and the use of contractile machinery at
the surface to allow for vesicle pinching.(3) Apoptotic bodies are released by dying cells in the final stage of apoptosis.
Compared to the other extracellular vesicles, the diameter of apoptotic bodies is the largest from 50nm to 5µm. (4).
EVs have been identified as important signaling mediators in a myriad of physiological and pathological processes. EVs
carry biomolecules from one cell to another, exposing the molecular state of the cell of origin. EVs are emerging as
powerful tools for theranostic purposes as their sheer presence in biological fluids designates them as novel and easily
accessible sources for the diagnosis of diseases, prognosis and treatment monitoring. (5)
This report prioritizes EV-based diagnostics with translational significance to across salient tumor types (pancreatic,
prostate, breast, lung, colorectal) and hematologic settings, and across biofluids including plasma/serum and
urine, where biomarker validity, analytical performance, and clinical pragmatism are the priority (6, 7). EV biology
and interactions with the tumor microenvironment are considered only in contexts directly relevant to diagnostic
interpretation and clinical application—for example, stromal EV–mediated resistance mechanisms that overlap with

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