Abstract
Glycosylation is highly sensitive to the biochemical environment and plays a key role in development and disease manifestation. Moreover, glycan biosynthesis depends on several highly competitive processes; thus, variations in the concentration of specific glycosyltransferases produce different products. For this reason, monitoring changes in glycosylation may be a more specific and sensitive approach to biomarker discovery and possibly disease diagnosis. Glycans in serum are of particular interest as approximately half of all proteins are glycosylated. We have developed the methods for profiling the glycans in human serum to identify glycan biomarker. Global release methods were used including chemical and enzymatic to access O-linked and N-linked glycans, respectively. Glycans were released from the culture medium of various cancer cell lines, in control sera, and in cancer patients and isolated using solid phase extraction (SPE) with a porous graphitized carbon. The SPE fractions were analyzed by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI FTICR MS). Glycan compositions were determined based on accurate masses and tandem mass spectrometry. Glycosylation changes between control and patient group were monitored. Several glycans were identified as potential markers for ovarian, breast, and prostate cancer. In short, direct glycan analysis of human serum without any protein identification represents a new and innovative approach to disease marker discovery.
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Brockhausen, I. (1999) Pathways of O-glycan biosynthesis in cancer cells. Biochim. Biophys. Acta 1473, 67–95.
Dube, D. H. and Bertozzi, C. R. (2005) Glycans in cancer and inflammation. Potential for therapeutics and diagnostics. Nat. Rev. Drug Discov. 4, 477–488.
Fuster, M. M. and Esko, J. D. (2005) The sweet and sour of cancer: glycans as novel therapeutic targets. Nat. Rev. Cancer 5, 526–542.
Apweiler, R., Hermjakob, H., and Sharon, N. (1999) On the frequency of protein glycosylation, as deduced from analysis of the SWISS-PROT database. Biochim. Biophys. Acta 1473, 4–8.
Dennis, J. W., Granovsky, M., and Warren, C. E. (1999) Protein glycosylation in development and disease. Bioessays 21, 412–421.
Manning, J. C., Seyrek, K., Kaltner, H., Andre, S., Sinowatz, F., and Gabius, H. J. (2004) Glycomic profiling of developmental changes in bovine testis by lectin histochemistry and further analysis of the most prominent alteration on the level of the glycoproteome by lectin blotting and lectin affinity chromatography. Histol. Histopathol. 19, 1043–1060.
Thaysen-Andersen, M., Thogersen, I. B., Lademann, U., Offenberg, H., Giessing, A. M. B., Enghild, J. J., Nielsen, H. J., Brunner, N., and Hojrup, P. (2008) Investigating the biomarker potential of glycoproteins using comparative glycoprofiling – application to tissue inhibitor of metalloproteinases-1. Biochim. Biophys. Acta 1784, 455–463.
Uchiyama, N., Kuno, A., Tateno, H., Kubo, Y., Mizuno, M., Noguchi, M., and Hirabayashi, J. (2008) Optimization of evanescent-field fluorescence-assisted lectin microarray for high-sensitivity detection of monovalent oligosaccharides and glycoproteins. Proteomics 8, 3042–3050.
Kuno, A., Uchiyama, N., Koseki-Kuno, S., Ebe, Y., Takashima, S., Yamada, M., and Hirabayashi, J. (2005) Evanescent-field fluorescence-assisted lectin microarray: a new strategy for glycan profiling. Nat. Methods 2, 851–856.
Patwa, T. H., Zhao, J., Anderson, M. A., Simeone, D. M., and Lubman, D. M. (2006) Screening of glycosylation patterns in serum using natural glycoprotein microarrays and multi-lectin fluorescence detection. Anal. Chem. 78, 6411–6421.
An, H. J., Miyamoto, S., Lancaster, K. S., Kirmiz, C., Li, B. S., Lam, K. S., Leiserowitz, G. S., and Lebrilla, C. B. (2006) Profiling of glycans in serum for the discovery of potential biomarkers for ovarian cancer. J. Proteome Res. 5, 1626–1635.
Kirmiz, C., Li, B. S., An, H. J., Clowers, B. H., Chew, H. K., Lam, K. S., Ferrige, A., Alecio, R., Borowsky, A. D., Sulaimon, S., Lebrilla, C. B., and Miyamoto, S. (2007) A serum glycomics approach to breast cancer biomarkers. Mol. Cell. Proteomics 6, 43–55.
Kyselova, Z., Mechref, Y., Al Bataineh, M. M., Dobrolecki, L. E., Hickey, R. J., Vinson, J., Sweeney, C. J., and Novotny, M. V. (2007) Alterations in the serum glycome due to metastatic prostate cancer. J. Proteome Res. 6, 1822–1832.
de Leoz, M. L. A., An, H. J., Kronewitter, S., Kim, J., Beecroft, S., Vinall, R., Miyamoto, S., de Vere White, R., Lam, K. S., and Lebrilla, C. B. (2008) Glycomic approach for potential biomarkers on prostate cancer: profiling of N-linked glycans in human sera and pRNS cell lines. Dis. Markers: Glycobiol. 25, 243–258.
Kita, Y., Miura, Y., Furukawa, J., Nakano, M., Shinohara, Y., Ohno, M., Takimoto, A., and Nishimura, S. (2007) Quantitative glycomics of human whole serum glycoproteins based on the standardized protocol for liberating N-glycans. Mol. Cell. Proteomics 6, 1437–1445.
Morelle, W., Flahaut, C., Michalski, J. C., Louvet, A., Mathurin, P., and Klein, A. (2006) Mass spectrometric approach for screening modifications of total serum N-glycome in human diseases: application to cirrhosis. Glycobiology 16, 281–293.
Kam, R. K. T., Poon, T. C. W., Chan, H. L. Y., Wong, N., Hui, A. Y., and Sung, J. J. Y. (2007) High-throughput quantitative profiling of serum N-glycome by MALDI-TOF mass spectrometry and N-glycomic fingerprint of liver fibrosis. Clin. Chem. 53, 1254–1263.
Harvey, D. J., Royle, L., Radcliffe, C. M., Rudd, P. M., and Dwek, R. A. (2008) Structural and quantitative analysis of N-linked glycans by matrix-assisted laser desorption ionization and negative ion nanospray mass spectrometry. Anal. Biochem. 376, 44–60.
Isailovic, D., Kurulugama, R. T., Plasencia, M. D., Stokes, S. T., Kyselova, Z., Goldman, R., Mechref, Y., Novotny, M. V., and Clemmer, D. E. (2008) Profiling of human serum glycans associated with liver cancer and cirrhosis by IMS-MS. J. Proteome Res. 7, 1109–1117.
Zhao, J., Qiu, W. L., Simeone, D. M., and Lubman, D. M. (2007) N-linked glycosylation profiling of pancreatic cancer serum using capillary liquid phase separation coupled with mass spectrometric analysis. J. Proteome Res. 6, 1126–1138.
Lebrilla, C. B. and An, H. J. (2009) The prospects of glycan biomarkers for the diagnosis of diseases. Mol. Biosyst. 5, 17–20.
Leiserowitz, G. S., Lebrilla, C., Miyamoto, S., An, H. J., Duong, H., Kirmiz, C., Ll, B., Liu, H., and Lam, K. S. (2008) Glycomics analysis of serum: a potential new biomarker for ovarian cancer? Int. J. Gynecol. Cancer 18, 470–475.
Li, B. S., An, H. J., Kirmiz, C., Lebrilla, C. B., Lam, K. S., and Miyamoto, S. (2008) Glycoproteomic analyses of ovarian cancer cell lines and sera from ovarian cancer patients show distinct glycosylation changes in individual proteins. J. Proteome Res. 7, 3776–3788.
Lebrilla, C. B. and An, H. J. (2008) The prospects of glycan biomarkers for the diagnosis of diseases. Mol. Biosyst. 5, 17–20.
Greiner, M., Pfeiffer, D., and Smith, R. D. (2000) Principles and practical application of the receiver-operating characteristic analysis for diagnostic tests. Prev. Vet. Med. 45, 23–41.
Barkauskas, D. A., An, H. J., Kronewitter, S. R., de Leoz, L. M., Chew, H. K., de Vere White, R. W., Leiserowitz, G. S., Miyamoto, S., Lebrilla, C. B., and Rocke, D. M. (2009) Detecting glycan cancer biomarkers in serum samples using MALDI FT-ICR mass spectrometry data. Bioinformatics, 25, 251–257.
Hopkins, A. M., Miller, C. J., Connolly, A. J., Genovese, C., Nichol, R. C., and Wasserman, L. (2002) A new source detection algorithm using the false-discovery rate. Astron. J. 123, 1086–1094.
Acknowledgments
The authors gratefully acknowledge the financial support by the National Institutes of Health (R01 GM049077).
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An, H.J., Lebrilla, C.B. (2010). A Glycomics Approach to the Discovery of Potential Cancer Biomarkers. In: Li, J. (eds) Functional Glycomics. Methods in Molecular Biology, vol 600. Humana Press. https://doi.org/10.1007/978-1-60761-454-8_14
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DOI: https://doi.org/10.1007/978-1-60761-454-8_14
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