EphA2 on urinary extracellular vesicles as a novel biomarker for bladder cancer diagnosis and its effect on the invasiveness of bladder cancer

• Published in: British journal of cancer 2022-10, Vol.127 (7), p.1312-1323
• Main Authors: Tomiyama, Eisuke, Fujita, Kazutoshi, Matsuzaki, Kyosuke, Narumi, Ryohei, Yamamoto, Akinaru, Uemura, Toshihiro, Yamamichi, Gaku, Koh, Yoko, Matsushita, Makoto, Hayashi, Yujiro, Hashimoto, Mamoru, Banno, Eri, Kato, Taigo, Hatano, Koji, Kawashima, Atsunari, Uemura, Motohide, Ukekawa, Ryo, Takao, Tetsuya, Takada, Shingo, Uemura, Hirotsugu, Adachi, Jun, Tomonaga, Takeshi, Nonomura, Norio
• Format: Article
• Language: English
• Subjects: 631/337/475; 631/67/1857; 631/67/2322; 631/67/589/1336; Biomarkers; Biomarkers - metabolism; Biomedical and Life Sciences; Biomedicine; Bladder cancer; Cancer; Cancer Research; CD9 antigen; Cell migration; Diagnosis; Drug Resistance; Enzyme-linked immunosorbent assay; EphA2 protein; Ephrins - metabolism; Epidemiology; Extracellular vesicles; Extracellular Vesicles - metabolism; Hematuria; Humans; Medical diagnosis; Membrane proteins; Molecular Medicine; Oncology; Patients; Proteins; Proteomics; Receptor, EphA2; Ultracentrifugation; Urinary Bladder Neoplasms - diagnosis; Urinary Bladder Neoplasms - metabolism
• ISSN: 0007-0920; 1532-1827
• DOI: 10.1038/s41416-022-01860-0

Background Urinary extracellular vesicles (uEVs) secreted from bladder cancer contain cancer-specific proteins that are potential diagnostic biomarkers. We identified and evaluated a uEV-based protein biomarker for bladder cancer diagnosis and analysed its functions. Methods Biomarker candidates, selected by shotgun proteomics, were validated using targeted proteomics of uEVs obtained from 49 patients with and 48 individuals without bladder cancer, including patients with non-malignant haematuria. We developed an enzyme-linked immunosorbent assay (ELISA) for quantifying the uEV protein biomarker without ultracentrifugation and evaluated urine samples from 36 patients with and 36 patients without bladder cancer. Results Thirteen membrane proteins were significantly upregulated in the uEVs from patients with bladder cancer in shotgun proteomics. Among them, eight proteins were validated by target proteomics, and Ephrin type-A receptor 2 (EphA2) was the only protein significantly upregulated in the uEVs of patients with bladder cancer, compared with that of patients with non-malignant haematuria. The EV-EphA2-CD9 ELISA demonstrated good diagnostic performance (sensitivity: 61.1%, specificity: 97.2%). We showed that EphA2 promotes proliferation, invasion and migration and EV-EphA2 promotes the invasion and migration of bladder cancer cells. Conclusions We established EV-EphA2-CD9 ELISA for uEV-EphA2 detection for the non-invasive early clinical diagnosis of bladder cancer.