Extracellular Particles to Track Cancer Biomarkers from Tissue to Biofluids

Liquid biopsies that analyze the molecular content of extracellular vesicles and particles (EVPs) are a formidable opportunity for early and late-stage cancer diagnosis. This study explores the potential of two advanced technologies: Bessel beam excitation separation technology (BEST) and multiparametric biochip assay (MBA), to track single EVP cargo from organs of pathology into biofluids such as plasma and saliva. Using gastric cancer (GC) as a disease model, we conducted high-throughput, multiparametric analyses of EVPs derived from plasma, saliva, and tissue samples from GC patients. Our findings demonstrate the feasibility of these techniques in isolating and characterizing EVPs, revealing consistent EVP morphology and size across biofluids. Furthermore, differential expression patterns of the developed and validated salivary GC biomarkers, miR-140-5p and miR-301a-3p, were observed in the biofluids of GC patients, supporting the diagnostic relevance of these cargo molecules. Notably, saliva emerged as the most promising biofluid for GC diagnosis, achieving superior receiver-operating characteristic curve values compared with plasma and tissue. This study highlights the role of BEST and MBA in advancing single-EVP analysis and elucidating EVP trafficking, paving the way for future diagnostic applications of EVP cargo.