Tanaka-Okamoto Miki, Hanzawa Ken, Yamamoto Takashi, Michida Tomoki, Ikezawa Kenji, Ohkawa Kazuyoshi, Nishino Kazumi, Yokota Takafumi, Maeda Michihide, Yoshida Ken-Ichi, Takenaka Satoshi, Ohue Masayuki, Yamasaki Tomoyuki, Miyamoto Yasuhide
Department of Glyco-Oncology and Medical Biochemistry, Osaka International Cancer Institute, 3-1-69 Otemae, Chuo-Ku, Osaka, 541-8567, Japan.
Glycoanalytical Chemistry Lab, Institute for Glyco-Core Research (iGCORE), Gifu University, 1-1 Yanagido, Gifu, 501-1193, Japan.
Sci Rep. 2025 Mar 24;15(1):10109. doi: 10.1038/s41598-025-94496-y.
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.
在我们之前的研究中鉴定出了许多尿游离聚糖标志物。在此，对这些标志物的临床实用性进行了研究。分析了从120名健康受试者和503名患有各种恶性肿瘤的患者采集的尿液样本。合成了四种含N-羟乙酰神经氨酸（GlcNGc）的乳糖核心聚糖并用作内标（ISs）。使用两步柱纯化程序分离游离聚糖，进行吡啶基胺化，然后进行液相色谱-选择反应监测。使用四种内标和八种参考聚糖进行分析方法验证。选择由三种唾液酸化乳糖核心聚糖、三种硫酸化聚糖、四种Gn1核心游离N-聚糖和两种具有异常结构的聚糖组成的12种标志物来研究它们在癌症诊断中的有效性。同时测量标志物并对相对面积比（标志物/内标）进行定量。通过对尿液中的校准品和质量控制样品进行重复分析，证明该方法准确且精密。受试者工作特征曲线分析表明，单个标志物不足以进行高度准确的诊断。然而，四种标志物的组合分别使胃癌、结直肠癌和胰腺癌的曲线下面积提高到0.910、0.867和0.914。此外，这些标志物的水平在各种其他恶性肿瘤中也升高。这些分析证明了游离聚糖标志物具有很高的临床实用性。
