Proteomic analysis of urinary and tissue‐exudative extracellular vesicles to discover novel bladder cancer biomarkers. Issue 5 (31st March 2021)

• Record Type: Journal Article
• Authors: Tomiyama, Eisuke; Matsuzaki, Kyosuke; Fujita, Kazutoshi; Shiromizu, Takashi; Narumi, Ryohei; Jingushi, Kentaro; Koh, Yoko; Matsushita, Makoto; Nakano, Kosuke; Hayashi, Yujiro; Wang, Cong; Ishizuya, Yu; Kato, Taigo; Hatano, Koji; Kawashima, Atsunari; Ujike, Takeshi; Uemura, Motohide; Takao, Tetsuya; Adachi, Jun; Tomonaga, Takeshi; Nonomura, Norio
• Is Part Of: Cancer science. Volume 112:Issue 5(2021)
• Page Start: 2033
• Page End: 2045
• Publication Date: 2021-03-31
• Subjects: bladder cancer -- exosome -- extracellular vesicle -- proteomics -- urinary biomarker; Cancer -- Periodicals; Neoplasms -- Periodicals; Research -- Periodicals; Electronic journals; 616.994005
• DOI: 10.1111/cas.14881
• Languages: English
• ISSNs: 1347-9032

Abstract

Abstract: Proteomic analysis of urinary extracellular vesicles (EVs) is a powerful approach to discover potential bladder cancer (BCa) biomarkers, however urine contains numerous EVs derived from the kidney and normal urothelial epithelium, which can obfuscate information related to BCa cell‐derived EVs. In this study, we combined proteomic analysis of urinary EVs and tissue‐exudative EVs (Te‐EVs), which were isolated from culture medium of freshly resected viable BCa tissues. Urinary EVs were isolated from urine samples of 11 individuals (7 BCa patients and 4 healthy individuals), and Te‐EVs were isolated from 7 BCa tissues. We performed tandem mass tag (TMT)‐labeling liquid chromatography (LC‐MS/MS) analysis for both urinary EVs and Te‐EVs and identified 1960 proteins in urinary EVs and 1538 proteins in Te‐EVs. Most of the proteins identified in Te‐EVs were also present in urinary EVs (82.4%), with 55 of these proteins showing upregulated levels in the urine of BCa patients (fold change > 2.0; P < .1). Among them, we selected 22 membrane proteins as BCa biomarker candidates for validation using selected reaction monitoring/multiple reaction monitoring (SRM/MRM) analysis on urine samples from 70 individuals (40 BCa patients and 30 healthy individuals). Six urinary EV proteins (heat‐shock protein 90, syndecan‐1, myristoylated alanine‐rich C‐kinase substrate (MARCKS), MARCKS‐related protein, tight junction protein ZO‐2, and complement decay‐accelerating factor) were quantified using SRM/MRM analysis and validated as significantly upregulated in BCa patients ( P < .05). In conclusion, the novel strategy that combined proteomic analysis of urinary EVs and Te‐EVs enabled selective detection of urinary BCa biomarkers. Abstract : Proteomic analysis of urinary extracellular vesicles (EVs) is a powerful approach to discovering potential BCa biomarkers, however urine contains numerous EVs derived from kidney and normal urothelial epithelium that could dilute the information of cancer BCa cell‐derived EVs. In this study, we performed combined proteomic analysis of both urinary EVs and tissue‐extracted EVs (Te‐EVs) to identify reliable BCa biomarkers. This novel strategy presented here identified reliable urinary EV biomarker proteins exhibiting high levels of specificity and sensitivity for non‐invasive BCa detection.