Novel Glycobiomarker for Ovarian Cancer That Detects Clear Cell Carcinoma

Abstract

Epithelial ovarian cancer (EOC) is often asymptomatic and thus diagnosed at advanced stages with a poor prognosis. False-negative results for the conventional marker CA125 frequently occur in cases of clear cell carcinoma (CCC), a type of EOC; therefore, it is necessary to develop biomarkers with greater sensitivity. We previously reported a strategy to discover glycobiomarker candidates by combined lectin microarray and IGOT-LC/MS analysis. We have now optimized this strategy for discovering EOC biomarkers. Glycopeptides possessing cancerous glycans were enriched from the ascites fluids and culture supernatants of cancer cell lines with a fucose-binding lectin, AAL. IGOT-LC/MS analysis of CCC samples yielded 144 candidate glycoproteins. We selected WFA by lectin microarray as the optimal lectin to distinguish EOC from gastric and colon cancer. The candidates were narrowed by Western analysis of the WFA-bound fraction of ascites fluids. One of the final candidates, WFA-reactive ceruloplasmin, produced higher signals in the ascites fluids of EOC patients, including CCC, in comparison with the benign samples, while CA125 levels were comparable in the sandwich ELISA. Thus, our glycoproteomic strategy featuring efficient enrichment of glycans with disease-related alterations is applicable to various diseases.

Wisteria floribunda Agglutinin and Its Reactive-Glycan-Carrying Prostate-Specific Antigen as a Novel Diagnostic and Prognostic Marker of Prostate Cancer

“…In contrast, the enhanced expression of LacdiNAc has been shown to be associated with the progression of human prostate, ovarian, colon, and liver cancers [12,15,16,17]. Therefore, the quantification of LacdiNAc glycan carrying glycoproteins or tissue-specific expression of LacdiNAc glycan detected by the WFA has shown promise as cancer glycobiomarkers [17,18,19]. In particular, regarding PCa, there are only three papers about LacdiNAc distribution in prostate biopsy (Pbx) and RP specimens using WFA [15,16,20], and they did not report the relation between WFA-reactivity in tissues and PCa prognosis.…”

Application of Lectin Microarrays for Biomarker Discovery

“…The EOC‐specific lectin Wisteria floribunda agglutinin (WFA) was used for subsequent Western blot analysis to elucidate glycobiomarker candidates from the WFA‐bound fraction of ascites fluids. The WFA‐reactive ceruloplasmin generated higher signals in the ascites fluids of EOC patients (Table ) . Zhao et al.…”

“…The WFA-reactive ceruloplasmin generated higher signals in the ascites fluids of EOC patients (Table 3). [106] Zhao et al performed lectin microarray-MS glycomic analysis of ovarian cancer side population cells to determine antigens associated with cancer recurrence and drug resistance. They observed increased core fucosylated N-glycan and tumorassociated Tn, T, and sT antigen levels but decreased hybrid glycan, α2,3-linked sialic glycan, and multivalent sialylÀ glycan levels in side population cells.…”

Development of M2BPGi: a novel fibrosis serum glyco-biomarker for chronic hepatitis/cirrhosis diagnostics

“…The glycan profile of the selected glycoprotein was analyzed by lectin microarray, and the most appropriate lectin to be used in the serum test was selected. The selected glycoprotein was regarded as the target glycoprotein and the detection system for its glycan alteration was established based on the lectin-antibody sandwich assay system [46,47]. Glyco-pathology drug delivery system technology [72,73] Kikuchi et al (2006) Bioinformatics database (JCGGDB) [74] Togayachi et al Glycogene-modified cells/ mice (KO, Tg mice) [75][76][77] Ito et al (2009) Glycogene qPCR array [78] IGOT: Isotope-coded glycosylation site-specific tagging-liquid chromatography; JCGGDB: Japan consortium for glycobiology and glycotechnology database; SE: Sulfate emerging; SMME: Supported molecular matrix electrophoresis.…”

“…For instance, lung cancer can be classified into small cell carcinoma, adenocarcinoma, squamous cell carcinoma and large cell carcinoma, while ovarian cancer is classified into clear cell carcinoma, mucinous adenocarcinoma, serous adenocarcinoma and endometrioid adenocarcinoma. If the histological types of cancer in the same organ are different, glycan structures and probing lectins are largely different and distinctive [14,46]. Therefore, a collaborative study with experienced clinical doctors and pathologists is essential.…”