Expression of Heart Development Protein With EGF-Like Domains 1 (HEG1) Decorated With Low-Sulfated Keratan Sulfate in Human Malignant Pleural Mesothelioma.

IF 3.4 4区医学 Q2 PATHOLOGY

Pathology International Pub Date : 2025-06-17 DOI:10.1111/pin.70033

Koki Nakashima, Hitomi Hoshino, Zui Zhang, Tomoya O Akama, Nobuyuki Kondo, Seiki Hasegawa, Yoshitaka Sekido, Mana Fukushima, Tamotsu Ishizuka, Motohiro Kobayashi

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Abstract

The glycoform of heart development protein with EGF-like domains 1 (HEG1) recognized by the SKM9-2 monoclonal antibody is a useful diagnostic marker for malignant pleural mesothelioma (MPM). The putative glycoform includes core 2 O-glycans carrying sialyl poly-N-acetyllactosamine (LacNAc), but sulfation modifications are undetermined. Since sialyl 6-sulfo LacNAc-capped core 2 O-glycans are expressed in MPM and their structure overlaps with low-sulfated keratan sulfate (KS), we asked whether low-sulfated KS is expressed in MPM and whether HEG1 is decorated with low-sulfated KS. We performed immunohistochemical analysis of 41 MPM cases using anti-KS monoclonal antibodies and endoglycosidases, reversed-phase ion-pair high-performance liquid chromatography analysis of KS/sulfated LacNAc isolated from human pleural tissue, and western blot analysis of HEG1·IgG recombinant fusion proteins secreted from low-sulfated KS-expressing human embryonic kidney cells. Most MPM tissues were stained with anti-low-sulfated KS antibodies and staining was eliminated by endo-β-galactosidase and keratanase II but not by peptide-N-glycosidase F. Disaccharide composition analysis revealed that mono-sulfated LacNAc disaccharide and di-sulfated LacNAc disaccharide accounted for 83.1% and 16.9% of pleural KS/sulfated LacNAc, respectively. Western blot analysis of HEG1·IgG glycoforms indicated that HEG1 functions as a core protein for low-sulfated KS. Thus, HEG1 protein decorated with low-sulfated KS is expressed in MPM.

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低硫酸角蛋白修饰的egf样结构域1 （HEG1）在人恶性胸膜间皮瘤中的表达

SKM9-2单克隆抗体识别的具有egf样结构域1 （HEG1）的心脏发育蛋白糖型是恶性胸膜间皮瘤（MPM）的有用诊断标志物。假定的糖形式包括核心2 - o -聚糖携带唾液酰聚n -乙酰乳胺（LacNAc），但磺化修饰尚未确定。由于sialyl - 6-sulfo LacNAc-capped核心2 - o -甘聚糖在MPM中表达，且其结构与低硫酸化角蛋白硫酸盐（KS）重叠，我们想知道低硫酸化KS是否在MPM中表达，HEG1是否被低硫酸化KS修饰。我们使用抗KS单克隆抗体和内糖苷酶对41例MPM病例进行免疫组织化学分析，对从人胸膜组织分离的KS/硫酸盐LacNAc进行反相离子对高效液相色谱分析，并对低硫酸盐表达KS的人胚胎肾细胞分泌的HEG1·IgG重组融合蛋白进行western blot分析。大多数MPM组织用抗低硫化KS抗体染色，染色可被内切β-半乳糖苷酶和角化酶II消除，但不能被肽- n -糖苷酶f消除。双糖组成分析显示，单硫酸化LacNAc双糖和双硫酸化LacNAc双糖分别占胸膜KS/硫酸化LacNAc的83.1%和16.9%。Western blot分析HEG1·IgG糖型表明，HEG1是低硫KS的核心蛋白。因此，低硫化KS修饰的HEG1蛋白在MPM中表达。

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来源期刊

Pathology International 医学-病理学

CiteScore

4.50

自引率

4.50%

发文量

102

审稿时长

12 months

期刊介绍： Pathology International is the official English journal of the Japanese Society of Pathology, publishing articles of excellence in human and experimental pathology. The Journal focuses on the morphological study of the disease process and/or mechanisms. For human pathology, morphological investigation receives priority but manuscripts describing the result of any ancillary methods (cellular, chemical, immunological and molecular biological) that complement the morphology are accepted. Manuscript on experimental pathology that approach pathologenesis or mechanisms of disease processes are expected to report on the data obtained from models using cellular, biochemical, molecular biological, animal, immunological or other methods in conjunction with morphology. Manuscripts that report data on laboratory medicine (clinical pathology) without significant morphological contribution are not accepted.