Protein glycosylation is a ubiquitous and complex co- and post-translational modification leading to glycan formation, i.e. oligosaccharide chains covalently attached to peptide backbones. The significance of changes in glycosylation for the beginning, progress and outcome of different human diseases is widely recognized. Thus, glycans are considered as unique structures to diagnose, predict susceptibility to and monitor the progression of disease. In the »omics« era, the glycome, a glycan analogue of the proteome and genome, holds considerable promise as a source of new biomarkers. In the design of a strategy for biomarker discovery, new principles and platforms for the analysis of relatively small amounts of numerous glycoproteins are needed. Emerging glycomics technologies comprising different types of mass spectrometry and affinity-based arrays are next in line to deliver new analytical procedures in the field of biomarkers. Screening different types of glycomolecules, selection of differentially expressed components, their enrichment and purification or identification are the most challenging parts of experimental and clinical glycoproteomics. This requires large-scale technologies enabling high sensitivity, proper standardization and validation of the methods to be used. Further progress in the field of applied glycoscience requires an integrated systematic approach in order to explore properly all opportunities for disease diagnosis.
Evaluation of the Pattern of Human Serum Glycoproteins in Prostate Cancer
Glycoprotein profiling at the level of cells, tissues and biological fluids is aimed at discovering new cancer biomarkers and also at finding specific cancer-related structural alterations of known tumor markers. In this study we comparatively evaluated the glycoprotein patterns of human prostate cancer (PCa)- and normal human sera regarding sialylation and fucosylation as structural characteristics relevant for cancer progression. Glycoproteins were isolated using affinity chromatography on Sambucus nigra agglutinin- and Lens culinaris agglutinin-columns and subsequently characterized by SDS-PAGE and on-chip normal phase-surface capture combined with surface-enhanced laser/desorption ionization time of flight mass spectrometry. Comparative analysis of the glycoproteins purified from healthy and PCa sera indicated differences and redundancy of the isolated molecules in terms of the microheterogeneity of counterpart glycans, the relative abundance and the presence/absence of particular molecular species. In PCa there was a general increase in sialylation and decrease in fucosylation of human serum glycans compared to normal sera. Taken together, the results obtained indicated that an affinity-approach based on the use of lectins of narrow specificity reduced the complexity of the examined samples and at this discovery-phase of our study pointed to specific glyco-changes that may be relevant for improving the monitoring of PCa progression.
Glycans as a Target in the Detection of Reproductive Tract Cancers
The significance of changes in glycosylation for the beginning, progress and outcome of different human diseases is highly recognized. In this review we summarized literature data on the alteration of glycans in cancer, especially glycoforms of tumor markers of reproductive tract cancers: prostate-specific antigen (PSA) and cancer antigen 125 (CA125). We aimed to highlight the diagnostic potential and relevance of glycan microheterogeneity and to present some novel methods for cancer detection. A computerized search of articles published up to 2007 was performed through the PubMed database. Search terms utilized included prostate/ovarian cancer glycosylation, prostate/ovarian cancer detection, PSA/CA125 glycosylation. Additional sources were identified through cross-referencing and researching in available biomedical books. The comparative studies of sugar chain structures of the PSA and CA125 indicated specific structural alterations associated with malignant transformation, in relation to glycan branching, sialylation and fucosylation. These glycan modifications should be better in distinguishing between benign and malignant conditions than the measurement of marker concentrations alone, which is widely used in practice. Cancer-associated changes in the glycosylation could yield more sensitive and discriminative diagnostic tests for reproductive tract cancer detection, i.e. for improvement of the clinical utility of known tumor markers or the discovery of new ones.
Background: Prostate-specific antigen (PSA) is a glycoprotein tumor marker known to exist as numerous glycospecies. Investigations on its glycobiochemical properties aimed at their use in the preparation of adjuncts in determining PSA concentration for clinical purposes have accumulated a lot of data on its structural properties. In this study, we reconsidered unexplored ubiquitously present low molecular mass species of PSA regarding to molecular mass, origin and pathophysiological source specificity in order to evaluate them as biomarkers. Methods: Data on low molecular mass PSA-immunoreactive species from sera of subjects with prostate cancer (PCa), benign prostatic hyperplasia (BPH), breast cancer (BCa), and urine of healthy males obtained by on-chip immunoaffinity chromatography combined with mass spectrometry were analyzed. Results: The results obtained indicated PSA species common to BCa, PCa, and BPH at 12-13 kDa, 17-19 kDa and 21-24 kDa. The striking difference in predominant frequencies made the profile characteristic in each examined pathophysiological condition. On the other hand, paired groups of prostatic and extraprostatic PSA contained rare species with small differences among groups concerning individual species. Low molecular mass PSA also included rare species unique for each group of samples. Conclusion: The results obtained revealed that uniformity of low molecular mass PSA-immunoreactive species in sera prevails over diversity related to cancer and non-cancer conditions, but at the same time some of them are molecules with biomarker potential for BPH detection.
CA-125 (coelomic epithelium-related antigen) forms the extracellular portion of transmembrane mucin 16 (MUC16). It is shed after proteolytic degradation. Due to structural heterogeneity, CA-125 ligand capacity and biological roles are not yet understood. In this study, we assessed CA-125 as a ligand for dendritic cell-specific ICAM-3-grabbing non-integrin (DC-SIGN), which is a C-type lectin showing specificity for mannosylated and fucosylated structures. It plays a role as a pattern recognition molecule for viral and bacterial glycans or as an adhesion receptor. We probed a human DC-SIGN-Fc chimera with CA-125 of fetal or cancer origin using solid- or fluid-phase binding and inhibition assays. The results showed that DC-SIGN binds to CA-125 of fetal origin and that this interaction is carbohydrate-dependent. By contrast, cancerderived CA-125 displayed negligible binding. Inhibition assays indicated differences in the potency of CA-125 to interfere with DC-SIGN binding to pathogen-related glycoconjugates, such as mannan and Helicobacter pylori antigens. The differences in ligand properties between CA-125 of fetal and cancer origin may be due to specificities of glycosylation. This might influence various functions of dendritic cells based on their subset diversity and maturation-related functional capacity.