Association between Urinary-Extracellular-Vesicle-Enriched Proteome Dynamics and Oncological Outcomes Following...

Although concurrent chemoradiation (CCRT) is the standard of care for locally advanced cervical cancer, oncological outcomes remain suboptimal. Noninvasive monitoring of treatment responses could improve therapeutic management. This study investigated the temporal dynamics of extracellular vesicle (EV)-enriched urinary proteomes in patients with locally advanced cervical cancer undergoing CCRT to identify physiobiological shifts and proteins associated with treatment response and survival. Urine samples from 42 patients were collected longitudinally before, 1 month post-, and 3 months post-CCRT completion (126 samples). Urinary EVs were enriched using strong-anion-exchange magnetic beads. Proteomic profiling was performed using liquid chromatography-tandem mass spectrometry. Differential protein abundance and time-varying Cox regression analyses were used to correlate the proteomic signatures with CCRT response and overall survival. Analysis of longitudinal samples yielded 2,352 quantifiable proteins. Of these, 1,055 exhibited significant temporal shifts. Pathway analysis revealed that tumors and virus-associated proteins peaked at 3 months posttreatment, corresponding to the clinical response assessment window. By 3 months, nonresponders exhibited pronounced suppression of adaptive immune pathways and up-regulation of protein homeostasis and degradation pathways. Time-varying survival analysis identified 8 proteins driven by an altered T-complex ring complex/chaperonin containing T-complex protein 1, ribosomal proteins, and immune-related proteins associated with overall survival. Six of these were expressed in cervical tumor tissues based on the Human Protein Atlas. Overall, EV-enriched urinary proteomics provides a noninvasive liquid biopsy method for monitoring biological dynamics during CCRT. The observed immune suppression in nonresponders and the 8-protein prognostic signature offer preliminary insight into survival patterns and could help guide future studies on adjuvant immune-enhancing approaches.