Combination of urinary free-glycan markers for the diagnosis of various malignant tumors

Abstract Many urinary free-glycan markers were identified in our previous studies. Here, the clinical utility of these markers was examined. Urine samples taken from 120 healthy subjects and 503 patients with various malignant tumors were analyzed. Four lactose-core glycans containing N-glycolylgluc...

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Main Authors: Miki Tanaka-Okamoto, Ken Hanzawa, Takashi Yamamoto, Tomoki Michida, Kenji Ikezawa, Kazuyoshi Ohkawa, Kazumi Nishino, Takafumi Yokota, Michihide Maeda, Ken-ichi Yoshida, Satoshi Takenaka, Masayuki Ohue, Tomoyuki Yamasaki, Yasuhide Miyamoto
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Language:English
Published: Nature Portfolio 2025-03-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-94496-y
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author Miki Tanaka-Okamoto
Ken Hanzawa
Takashi Yamamoto
Tomoki Michida
Kenji Ikezawa
Kazuyoshi Ohkawa
Kazumi Nishino
Takafumi Yokota
Michihide Maeda
Ken-ichi Yoshida
Satoshi Takenaka
Masayuki Ohue
Tomoyuki Yamasaki
Yasuhide Miyamoto
author_facet Miki Tanaka-Okamoto
Ken Hanzawa
Takashi Yamamoto
Tomoki Michida
Kenji Ikezawa
Kazuyoshi Ohkawa
Kazumi Nishino
Takafumi Yokota
Michihide Maeda
Ken-ichi Yoshida
Satoshi Takenaka
Masayuki Ohue
Tomoyuki Yamasaki
Yasuhide Miyamoto
author_sort Miki Tanaka-Okamoto
collection DOAJ
description Abstract Many urinary free-glycan markers were identified in our previous studies. Here, the clinical utility of these markers was examined. Urine samples taken from 120 healthy subjects and 503 patients with various malignant tumors were analyzed. Four lactose-core glycans containing N-glycolylglucosamine (GlcNGc) were synthesized and used as internal standards (ISs). Free-glycans were isolated using a two-step column purification procedure, pyridylaminated, and subjected to LC-selected reaction monitoring. Assay validation was performed using four ISs and eight reference glycans. Twelve markers composed of three sialyl lactose-core glycans, three sulfated glycans, four Gn1-core free-N-glycans, and two glycans with unusual structures were selected to investigate their effectiveness for cancer diagnosis. Markers were simultaneously measured and the relative area ratio (marker/IS) quantified. This method was shown to be accurate and precise by repeated analysis of calibrators and quality control samples in urine. Receiver operating characteristic curve analyses revealed that individual markers were not sufficient for highly accurate diagnosis. However, a combination of four markers resulted in higher area under curves of 0.910, 0.867, and 0.914 for gastric, colorectal, and pancreatic cancers, respectively. Moreover, levels of these markers were elevated in various other malignancies. These analyses demonstrated high clinical utility of the free-glycan markers.
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spelling doaj-art-aa95acf528e9454cac42b204bdba883e2025-08-20T03:40:44ZengNature PortfolioScientific Reports2045-23222025-03-0115111110.1038/s41598-025-94496-yCombination of urinary free-glycan markers for the diagnosis of various malignant tumorsMiki Tanaka-Okamoto0Ken Hanzawa1Takashi Yamamoto2Tomoki Michida3Kenji Ikezawa4Kazuyoshi Ohkawa5Kazumi Nishino6Takafumi Yokota7Michihide Maeda8Ken-ichi Yoshida9Satoshi Takenaka10Masayuki Ohue11Tomoyuki Yamasaki12Yasuhide Miyamoto13Department of Glyco-Oncology and Medical Biochemistry, Osaka International Cancer InstituteGlycoanalytical Chemistry Lab, Institute for Glyco-Core Research (iGCORE), Gifu UniversityDepartment of Clinical Laboratory, Osaka International Cancer InstituteDepartment of Gastrointestinal Oncology, Osaka International Cancer InstituteDepartment of Hepatobiliary and Pancreatic Oncology, Osaka International Cancer InstituteDepartment of Hepatobiliary and Pancreatic Oncology, Osaka International Cancer InstituteDepartment of Thoracic Oncology, Osaka International Cancer InstituteDepartment of Hematology, Osaka International Cancer InstituteDepartment of Gynecology, Osaka International Cancer InstituteDepartment of Diagnostic Pathology and Cytology, Osaka International Cancer InstituteDepartment of Orthopedic Surgery, Osaka International Cancer InstituteDepartment of Gastroenterological Surgery, Osaka International Cancer InstituteDepartment of Clinical Laboratory, Osaka International Cancer InstituteDepartment of Clinical Laboratory, Osaka International Cancer InstituteAbstract Many urinary free-glycan markers were identified in our previous studies. Here, the clinical utility of these markers was examined. Urine samples taken from 120 healthy subjects and 503 patients with various malignant tumors were analyzed. Four lactose-core glycans containing N-glycolylglucosamine (GlcNGc) were synthesized and used as internal standards (ISs). Free-glycans were isolated using a two-step column purification procedure, pyridylaminated, and subjected to LC-selected reaction monitoring. Assay validation was performed using four ISs and eight reference glycans. Twelve markers composed of three sialyl lactose-core glycans, three sulfated glycans, four Gn1-core free-N-glycans, and two glycans with unusual structures were selected to investigate their effectiveness for cancer diagnosis. Markers were simultaneously measured and the relative area ratio (marker/IS) quantified. This method was shown to be accurate and precise by repeated analysis of calibrators and quality control samples in urine. Receiver operating characteristic curve analyses revealed that individual markers were not sufficient for highly accurate diagnosis. However, a combination of four markers resulted in higher area under curves of 0.910, 0.867, and 0.914 for gastric, colorectal, and pancreatic cancers, respectively. Moreover, levels of these markers were elevated in various other malignancies. These analyses demonstrated high clinical utility of the free-glycan markers.https://doi.org/10.1038/s41598-025-94496-yCancerFree-glycanMarkerSRMUrine
spellingShingle Miki Tanaka-Okamoto
Ken Hanzawa
Takashi Yamamoto
Tomoki Michida
Kenji Ikezawa
Kazuyoshi Ohkawa
Kazumi Nishino
Takafumi Yokota
Michihide Maeda
Ken-ichi Yoshida
Satoshi Takenaka
Masayuki Ohue
Tomoyuki Yamasaki
Yasuhide Miyamoto
Combination of urinary free-glycan markers for the diagnosis of various malignant tumors
Scientific Reports
Cancer
Free-glycan
Marker
SRM
Urine
title Combination of urinary free-glycan markers for the diagnosis of various malignant tumors
title_full Combination of urinary free-glycan markers for the diagnosis of various malignant tumors
title_fullStr Combination of urinary free-glycan markers for the diagnosis of various malignant tumors
title_full_unstemmed Combination of urinary free-glycan markers for the diagnosis of various malignant tumors
title_short Combination of urinary free-glycan markers for the diagnosis of various malignant tumors
title_sort combination of urinary free glycan markers for the diagnosis of various malignant tumors
topic Cancer
Free-glycan
Marker
SRM
Urine
url https://doi.org/10.1038/s41598-025-94496-y
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