TY - JOUR
T1 - Extracellular vesicles expressing CEACAM proteins in the urine of bladder cancer patients
AU - Igami, Ko
AU - Uchiumi, Takeshi
AU - Shiota, Masaki
AU - Ueda, Saori
AU - Tsukahara, Shigehiro
AU - Akimoto, Masaru
AU - Eto, Masatoshi
AU - Kang, Dongchon
N1 - Funding Information:
We thank the laboratory members of Clinical Chemistry and Laboratory Medicine, Kyushu University, for reagents, discussions, and carefully reviewing the manuscript. We would like to express our gratitude to the Department of Urology and Laboratory Medicine, Kyushu University, for collecting valuable clinical samples. We are grateful to LSI Medience Corporation for its generous support during this study. This research was supported by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (grant numbers 17H01550, 18 K15421, and 20H00530) and AMED (grant number JP21lm0203009). Finally, we thank H. Nikki March, PhD, from Edanz for editing a draft of this manuscript.
Funding Information:
We thank the laboratory members of Clinical Chemistry and Laboratory Medicine, Kyushu University, for reagents, discussions, and carefully reviewing the manuscript. We would like to express our gratitude to the Department of Urology and Laboratory Medicine, Kyushu University, for collecting valuable clinical samples. We are grateful to LSI Medience Corporation for its generous support during this study. This research was supported by Grants‐in‐Aid for Scientific Research from the Japan Society for the Promotion of Science (grant numbers 17H01550, 18 K15421, and 20H00530) and AMED (grant number JP21lm0203009). Finally, we thank H. Nikki March, PhD, from Edanz for editing a draft of this manuscript.
Publisher Copyright:
© 2022 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.
PY - 2022
Y1 - 2022
N2 - Early detection and long-term monitoring are important for urothelial carcinoma of the bladder (UCB). Urine cytology and existing markers have insufficient diagnostic performance. Here, we examined medium-sized extracellular vesicles (EVs) in urine to identify specific markers for UCB and evaluated their usefulness as diagnostic material. To identify specific markers in urinary EVs derived from UCB, we undertook shotgun proteomics using urine from four UCB patients and four healthy subjects. Next, 29 healthy specimens, 18 noncancer specimens, and 33 UCB specimens, all from men, were analyzed for urinary EVs by flow cytometry to evaluate the diagnostic performance of UCB-specific EVs. Nanoparticle-tracking analysis indicated that the size of EVs extracted from urine was mostly <400 nm. By shotgun proteomics, we detected several proteins characteristic of UCB and found that carcinoembryonic antigen-related adhesion molecule (CEACAM) proteins were increased in patients. Flow cytometric analysis revealed that the degree of expression of CEACAM1, CEACAM5, and CEACAM6 proteins on the surface of EVs varied among patients. Extracellular vesicles expressing CEACAM proteins also expressed mucin 1, suggesting that they were derived from tumorigenic uroepithelial cells. The number of EVs expressing CEACAM1, 5, and 6 proteins was significantly increased in UCB (mean ± SD, 8.6 ± 13%) compared to non-UCB (0.69 ± 0.46) and healthy (0.46 ± 0.34) by flow cytometry. The results of receiver operating characteristic (ROC) analysis showed a good score of area under the ROC curve of 0.907. We identified EVs that specifically express CEACAM proteins in urine and have potential for diagnostic applications. These EVs are potential targets in a new liquid biopsy test for UCB patients.
AB - Early detection and long-term monitoring are important for urothelial carcinoma of the bladder (UCB). Urine cytology and existing markers have insufficient diagnostic performance. Here, we examined medium-sized extracellular vesicles (EVs) in urine to identify specific markers for UCB and evaluated their usefulness as diagnostic material. To identify specific markers in urinary EVs derived from UCB, we undertook shotgun proteomics using urine from four UCB patients and four healthy subjects. Next, 29 healthy specimens, 18 noncancer specimens, and 33 UCB specimens, all from men, were analyzed for urinary EVs by flow cytometry to evaluate the diagnostic performance of UCB-specific EVs. Nanoparticle-tracking analysis indicated that the size of EVs extracted from urine was mostly <400 nm. By shotgun proteomics, we detected several proteins characteristic of UCB and found that carcinoembryonic antigen-related adhesion molecule (CEACAM) proteins were increased in patients. Flow cytometric analysis revealed that the degree of expression of CEACAM1, CEACAM5, and CEACAM6 proteins on the surface of EVs varied among patients. Extracellular vesicles expressing CEACAM proteins also expressed mucin 1, suggesting that they were derived from tumorigenic uroepithelial cells. The number of EVs expressing CEACAM1, 5, and 6 proteins was significantly increased in UCB (mean ± SD, 8.6 ± 13%) compared to non-UCB (0.69 ± 0.46) and healthy (0.46 ± 0.34) by flow cytometry. The results of receiver operating characteristic (ROC) analysis showed a good score of area under the ROC curve of 0.907. We identified EVs that specifically express CEACAM proteins in urine and have potential for diagnostic applications. These EVs are potential targets in a new liquid biopsy test for UCB patients.
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U2 - 10.1111/cas.15438
DO - 10.1111/cas.15438
M3 - Article
C2 - 35611462
AN - SCOPUS:85132898505
SN - 1347-9032
JO - Cancer Science
JF - Cancer Science
ER -
