EIF2S1 in Urinary Extracellular Vesicles as a Novel Diagnostic Marker for Bladder Cancer

ABSTRACT Background Urinary extracellular vesicles (uEVs), directly secreted from bladder cancer (BCa) cells, harbor potential for biomarker discovery. Methods We performed proteomic analysis to explore and validate uEV‐based diagnostic markers for BCa, with a focus on cytoplasmic EV proteins. Among...

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Main Authors: Eisuke Tomiyama, Kazutoshi Fujita, Kyosuke Matsuzaki, Ryohei Narumi, Makoto Matsushita, Yujiro Hayashi, Mamoru Hashimoto, Taigo Kato, Koji Hatano, Atsunari Kawashima, Takafumi Minami, Tetsuya Takao, Shingo Takada, Hirotsugu Uemura, Jun Adachi, Takeshi Tomonaga, Norio Nonomura
Format: Article
Language:English
Published: Wiley 2025-05-01
Series:Cancer Medicine
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Online Access:https://doi.org/10.1002/cam4.70964
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author Eisuke Tomiyama
Kazutoshi Fujita
Kyosuke Matsuzaki
Ryohei Narumi
Makoto Matsushita
Yujiro Hayashi
Mamoru Hashimoto
Taigo Kato
Koji Hatano
Atsunari Kawashima
Takafumi Minami
Tetsuya Takao
Shingo Takada
Hirotsugu Uemura
Jun Adachi
Takeshi Tomonaga
Norio Nonomura
author_facet Eisuke Tomiyama
Kazutoshi Fujita
Kyosuke Matsuzaki
Ryohei Narumi
Makoto Matsushita
Yujiro Hayashi
Mamoru Hashimoto
Taigo Kato
Koji Hatano
Atsunari Kawashima
Takafumi Minami
Tetsuya Takao
Shingo Takada
Hirotsugu Uemura
Jun Adachi
Takeshi Tomonaga
Norio Nonomura
author_sort Eisuke Tomiyama
collection DOAJ
description ABSTRACT Background Urinary extracellular vesicles (uEVs), directly secreted from bladder cancer (BCa) cells, harbor potential for biomarker discovery. Methods We performed proteomic analysis to explore and validate uEV‐based diagnostic markers for BCa, with a focus on cytoplasmic EV proteins. Among the 1960 proteins identified by shotgun proteomics (tandem mass tag‐labeled liquid chromatography–tandem mass spectrometry [LC–MS/MS]) of uEVs from seven patients with BCa and four healthy individuals, 17 cytoplasmic EV proteins were significantly elevated in the patients' urine (fold change > 1.5; p < 0.05). These 17 proteins were subsequently validated using targeted proteomics (selected reaction monitoring/multiple reaction monitoring) using urine samples from 49 and 48 patients with and without BCa, respectively, including those with non‐BCa hematuria. Results Ten measurable EV proteins remained significantly elevated in the urine of patients with BCa, with EV‐EIF2S1 demonstrating the best diagnostic performance (area under the receiver operating characteristic [ROC] curve [AUC] [ROCAUC]: 0.83). Additionally, EV‐EIF2S1 distinguished patients with BCa from those without BCa and hematuria in a suitable manner (ROCAUC: 0.92). Functional analysis of EIF2S1 in the BCa cell lines (T24 and 5637) showed that EIF2S1 knockdown markedly inhibited cell proliferation and induced cell cycle arrest and apoptosis, suggesting its essentiality for BCa cell growth and survival. Conclusions This study identified EV‐EIF2S1 as a novel, uEV‐based BCa diagnostic marker and demonstrated its functional significance in BCa cell growth and survival.
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spelling doaj-art-384fab23ed704451b5371c8bf16bbeb82025-08-20T03:05:29ZengWileyCancer Medicine2045-76342025-05-011410n/an/a10.1002/cam4.70964EIF2S1 in Urinary Extracellular Vesicles as a Novel Diagnostic Marker for Bladder CancerEisuke Tomiyama0Kazutoshi Fujita1Kyosuke Matsuzaki2Ryohei Narumi3Makoto Matsushita4Yujiro Hayashi5Mamoru Hashimoto6Taigo Kato7Koji Hatano8Atsunari Kawashima9Takafumi Minami10Tetsuya Takao11Shingo Takada12Hirotsugu Uemura13Jun Adachi14Takeshi Tomonaga15Norio Nonomura16Department of Urology Osaka University Graduate School of Medicine Suita Osaka JapanDepartment of Urology Osaka University Graduate School of Medicine Suita Osaka JapanDepartment of Urology Osaka University Graduate School of Medicine Suita Osaka JapanLaboratory of Proteomics for Drug Discovery, Center for Drug Design Research National Institutes of Biomedical Innovation, Health and Nutrition Ibaraki Osaka JapanDepartment of Urology Osaka University Graduate School of Medicine Suita Osaka JapanDepartment of Urology Osaka University Graduate School of Medicine Suita Osaka JapanDepartment of Urology Kindai University Faculty of Medicine Osaka‐Sayama Osaka JapanDepartment of Urology Osaka University Graduate School of Medicine Suita Osaka JapanDepartment of Urology Osaka University Graduate School of Medicine Suita Osaka JapanDepartment of Urology Osaka University Graduate School of Medicine Suita Osaka JapanDepartment of Urology Kindai University Faculty of Medicine Osaka‐Sayama Osaka JapanDepartment of Urology Osaka General Medical Center Osaka Osaka JapanDepartment of Urology Osaka Police Hospital Osaka Osaka JapanDepartment of Urology Kindai University Faculty of Medicine Osaka‐Sayama Osaka JapanLaboratory of Proteomics for Drug Discovery, Center for Drug Design Research National Institutes of Biomedical Innovation, Health and Nutrition Ibaraki Osaka JapanLaboratory of Proteomics for Drug Discovery, Center for Drug Design Research National Institutes of Biomedical Innovation, Health and Nutrition Ibaraki Osaka JapanDepartment of Urology Osaka University Graduate School of Medicine Suita Osaka JapanABSTRACT Background Urinary extracellular vesicles (uEVs), directly secreted from bladder cancer (BCa) cells, harbor potential for biomarker discovery. Methods We performed proteomic analysis to explore and validate uEV‐based diagnostic markers for BCa, with a focus on cytoplasmic EV proteins. Among the 1960 proteins identified by shotgun proteomics (tandem mass tag‐labeled liquid chromatography–tandem mass spectrometry [LC–MS/MS]) of uEVs from seven patients with BCa and four healthy individuals, 17 cytoplasmic EV proteins were significantly elevated in the patients' urine (fold change > 1.5; p < 0.05). These 17 proteins were subsequently validated using targeted proteomics (selected reaction monitoring/multiple reaction monitoring) using urine samples from 49 and 48 patients with and without BCa, respectively, including those with non‐BCa hematuria. Results Ten measurable EV proteins remained significantly elevated in the urine of patients with BCa, with EV‐EIF2S1 demonstrating the best diagnostic performance (area under the receiver operating characteristic [ROC] curve [AUC] [ROCAUC]: 0.83). Additionally, EV‐EIF2S1 distinguished patients with BCa from those without BCa and hematuria in a suitable manner (ROCAUC: 0.92). Functional analysis of EIF2S1 in the BCa cell lines (T24 and 5637) showed that EIF2S1 knockdown markedly inhibited cell proliferation and induced cell cycle arrest and apoptosis, suggesting its essentiality for BCa cell growth and survival. Conclusions This study identified EV‐EIF2S1 as a novel, uEV‐based BCa diagnostic marker and demonstrated its functional significance in BCa cell growth and survival.https://doi.org/10.1002/cam4.70964bladder cancercytoplasmic proteinsextracellular vesiclesliquid biopsyproteomicsurinary markers
spellingShingle Eisuke Tomiyama
Kazutoshi Fujita
Kyosuke Matsuzaki
Ryohei Narumi
Makoto Matsushita
Yujiro Hayashi
Mamoru Hashimoto
Taigo Kato
Koji Hatano
Atsunari Kawashima
Takafumi Minami
Tetsuya Takao
Shingo Takada
Hirotsugu Uemura
Jun Adachi
Takeshi Tomonaga
Norio Nonomura
EIF2S1 in Urinary Extracellular Vesicles as a Novel Diagnostic Marker for Bladder Cancer
Cancer Medicine
bladder cancer
cytoplasmic proteins
extracellular vesicles
liquid biopsy
proteomics
urinary markers
title EIF2S1 in Urinary Extracellular Vesicles as a Novel Diagnostic Marker for Bladder Cancer
title_full EIF2S1 in Urinary Extracellular Vesicles as a Novel Diagnostic Marker for Bladder Cancer
title_fullStr EIF2S1 in Urinary Extracellular Vesicles as a Novel Diagnostic Marker for Bladder Cancer
title_full_unstemmed EIF2S1 in Urinary Extracellular Vesicles as a Novel Diagnostic Marker for Bladder Cancer
title_short EIF2S1 in Urinary Extracellular Vesicles as a Novel Diagnostic Marker for Bladder Cancer
title_sort eif2s1 in urinary extracellular vesicles as a novel diagnostic marker for bladder cancer
topic bladder cancer
cytoplasmic proteins
extracellular vesicles
liquid biopsy
proteomics
urinary markers
url https://doi.org/10.1002/cam4.70964
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