以尿液細胞外囊泡作為非侵入性液態切片預測上泌尿道尿路上皮癌之轉移

羅建雄¹、張靜卉²、王竹安^2,3、孫弘羽⁴、詹皓程¹

¹國立成功大學醫學院附設醫院 泌尿部; ²國立成功大學醫學院; ³國立成功大學醫學院 基礎醫學研究所; ⁴國立成功大學 生理學科暨研究所

Urinary Extracellular Vesicles as Non-Invasive Liquid Biopsies for Predicting Metastasis in Upper Tract Urothelial Carcinoma

Chien-Hsiung Lo¹, Ching-Hui Chang², Chu-An Wang^2,3, Hung-Yu Sun⁴, Hau-Chern Jan¹

¹ Department of Urology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan; ² College of Medicine, National Cheng Kung University; ³ Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University; ⁴ Department of Physiology, National Cheng Kung University, Tainan, Taiwan

Purpose

Upper tract urothelial carcinoma (UTUC) is an aggressive malignancy with high recurrence and metastasis rates even after radical surgery. Although UTUC is globally uncommon, its prevalence is markedly high in regions such as Taiwan and some Asian countries. Reliable non-invasive biomarkers are crucial but lacking. This study investigated urinary extracellular vesicles (uEVs) as a potential liquid biopsy for predicting metastatic progression in UTUC after radical nephroureterectomy (RNU).

Materials & Methods

Patients undergoing RNU for UTUC at our institution were included. Paired preoperative urine and tumor tissue samples were collected, and clinical data were retrospectively reviewed. uEVs were isolated and quantified by nanoparticle tracking analysis, while tumor RNA underwent next-generation sequencing (NGS). Proteomic profiling of uEVs was performed using LC–MS/MS, followed by transcriptomic correlation and pathway enrichment analyses.

Results

Among 40 patients, postoperative metastasis occurred in 56. Neither uEV concentration nor size differed between metastatic and non-metastatic groups. Proteomic analysis identified enrichment of EV-associated proteins—ANXA1, CD63, LAMP2, and PROM1—exclusively in metastatic urine. NGS confirmed upregulation of these genes in metastatic tumors. Pathway analysis revealed activation of the Akt signaling cascade, suggesting its involvement in metastatic progression.

Conclusions

uEVs represent a promising non-invasive biomarker platform for assessing metastatic potential in UTUC. Our integrative proteo-transcriptomic analysis indicates that uEV composition, rather than abundance, reflects tumor aggressiveness. The consistent upregulation of ANXA1, CD63, LAMP2, and PROM1, alongside Akt pathway activation, suggests a shared molecular mechanism driving metastasis. These findings highlight the translational potential of uEV-based assays for postoperative risk stratification, surveillance, and therapeutic targeting in UTUC.