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A pilot study of salivary N-glycome in HBV-induced chronic hepatitis, cirrhosis, and hepatocellular carcinoma

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Abstract

Hepatitis B is a potentially life-threatening liver infection caused by the hepatitis B virus (HBV), which can lead to chronic liver disease and put people at high risk of death from cirrhosis of the liver and liver cancer. However, little is known about the correlation of salivary N-linked glycans related to HBV-infected liver diseases. Here we investigated N-linked glycome in saliva from 200 subjects (50 healthy volunteers (HV), 40 HBV-infected patients (HB), 50 cirrhosis patients (HC), and 60 hepatocellular carcinoma patients (HCC) using MALDI-TOF/TOF-MS. Representative MS spectra of N-glycans with signal-to-noise ratios >6 were annotated using the GlycoWorkbench program. A total of 40, 47, 29, and 33 N-glycan peaks were identified and annotated from HV, HB, HC, and HCC groups, respectively. There were 15 N-glycan peaks (e.g., m/z 1647.587, 1688.613 and 2101.755) were present in all groups. Three N-glycan peaks (m/z 2596.925, 2756.962, and 2921.031) were unique in HV group, 2 N-glycan peaks (m/z 1898.676 and 1971.692) were unique in HB group, 5 N-glycan peaks (m/z 1954.677, 2507.914, 2580.930, 2637.952, and 3092.120) were unique in HC group, and 3 N-glycan peaks (m/z 2240.830, 2507.914, and 3931.338) were unique in HCC group. The proportion of fucosylated N-glycans was apparently increased in the HCC group (84.8%) than in any other group (73.1% ± 0.01), however, the proportion of sialylated N-glycans was decreased in HCC group (12.1%) than in any other group (17.23% ± 0.003). Our data provide pivotal information to distinguish between HBV-associated hepatitis, cirrhosis and HCC, and facilitate the discovery of biomarkers for HCC during its early stages based on precise alterations of N-linked glycans in saliva.

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References

  1. Who.int: Hepatitis B. http://www.who.int/mediacentre/factsheets/fs204/en/ (2016)

  2. Liu, J., Fan, D.: Hepatitis B in China. Lancet (London, England). 369(9573), 1582–1583 (2007)

    Article  Google Scholar 

  3. Pfaffe, T., Cooper-White, J., Beyerlein, P., Kostner, K., Punyadeera, C.: Diagnostic potential of saliva: current state and future applications. Clin. Chem. 57(5), 675–687 (2011)

    Article  CAS  PubMed  Google Scholar 

  4. Javaid, M.A., Ahmed, A.S., Durand, R., Tran, S.D.: Saliva as a diagnostic tool for oral and systemic diseases. J. Oral. Biol. Craniofac. Res. 6(1), 66–75 (2016)

    Article  PubMed  Google Scholar 

  5. Foley 3rd, J.D., Sneed, J.D., Steinhubl, S.R., Kolasa, J., Ebersole, J.L., Lin, Y., Kryscio, R.J., McDevitt, J.T., Campbell, C.L., Miller, C.S.: Oral fluids that detect cardiovascular disease biomarkers. Oral Surg Oral Med Oral Pathol Oral Radiol. 114(2), 207–214 (2012)

    Article  PubMed  PubMed Central  Google Scholar 

  6. Bencharit, S., Baxter, S.S., Carlson, J., Byrd, W.C., Mayo, M.V., Border, M.B., Kohltfarber, H., Urrutia, E., Howard-Williams, E.L., Offenbacher, S., Wu, M.C., Buse, J.B.: Salivary proteins associated with hyperglycemia in diabetes: a proteomic analysis. Mol. BioSyst. 9(11), 2785–2797 (2013)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Hu, S., Arellano, M., Boontheung, P., Wang, J., Zhou, H., Jiang, J., Elashoff, D., Wei, R., Loo, J.A., Wong, D.T.: Salivary proteomics for oral cancer biomarker discovery. Clin Cancer Res. 14(19), 6246–6252 (2008)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Zhang, L., Xiao, H., Karlan, S., Zhou, H., Gross, J., Elashoff, D., Akin, D., Yan, X., Chia, D., Karlan, B.: Discovery and preclinical validation of salivary transcriptomic and proteomic biomarkers for the non-invasive detection of breast cancer. PLoS One. 5(12), e15573–e15573 (2010)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Zhang, L., Xiao, H., Wong, D.T.: Salivary biomarkers for clinical applications. Mol Diagn Ther. 13(4), 245–259 (2009)

    Article  CAS  PubMed  Google Scholar 

  10. Li, D., Mallory, T., Satomura, S.: AFP-L3: a new generation of tumor marker for hepatocellular carcinoma. Clin Chim Acta. 313(1–2), 15–19 (2001)

    Article  CAS  PubMed  Google Scholar 

  11. Lebrilla, C.B., An, H.J.: The prospects of glycan biomarkers for the diagnosis of diseases. Mol. BioSyst. 5(1), 17–20 (2009)

    Article  CAS  PubMed  Google Scholar 

  12. Kratz, E.M., Waszkiewicz, N., Kaluza, A., Szajda, S.D., Zalewska-Szajda, B., Szulc, A., Zwierz, K., Ferens-Sieczkowska, M.: Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study. Alcohol Alcohol. 49(1), 23–30 (2014)

    Article  CAS  PubMed  Google Scholar 

  13. Kozak, R.P., Urbanowicz, P.A., Punyadeera, C., Reiding, K.R., Jansen, B.C., Royle, L., Spencer, D.I., Fernandes, D.L., Wuhrer, M.: Variation of human salivary O-Glycome. PLoS One. 11(9), e0162824 (2016)

    Article  PubMed  PubMed Central  Google Scholar 

  14. Qin, Y., Zhong, Y., Zhu, M., Dang, L., Yu, H., Chen, Z., Chen, W., Wang, X., Zhang, H., Li, Z.: Age- and sex-associated differences in the glycopatterns of human salivary glycoproteins and their roles against influenza a virus. J. Proteome Res. 12(6), 2742–2754 (2013)

    Article  CAS  PubMed  Google Scholar 

  15. Zhong, Y., Qin, Y., Yu, H., Yu, J., Wu, H., Lin, C., Zhang, P., Wang, X., Jia, Z., Guo, Y.: Avian influenza virus infection risk in humans with chronic diseases. Sci Rep. 5, 8971 (2014)

    Article  Google Scholar 

  16. Yang, G.L., Tian-Ran, M.A., Zheng, L.I.: Enrichment and characterization of Total N-linked Glycans from glycoproteins by ultrafiltration units and mass spectrometry. Prog Biochem Biophys. 41(4), 403–408 (2014)

    CAS  Google Scholar 

  17. Yang, G., Tan, Z., Lu, W., Guo, J., Yu, H., Yu, J., Sun, C., Qi, X., Li, Z., Guan, F.: Quantitative glycome analysis of N-glycan patterns in bladder cancer vs normal bladder cells using an integrated strategy. J. Proteome Res. 14(2), 639–653 (2015)

    Article  CAS  PubMed  Google Scholar 

  18. Yang, G., Cui, T., Wang, Y., Sun, S., Ma, T., Wang, T., Chen, Q., Li, Z.: Selective isolation and analysis of glycoprotein fractions and their glycomes from hepatocellular carcinoma sera. Proteomics. 13(9), 1481–1498 (2013)

    Article  CAS  PubMed  Google Scholar 

  19. Tan, Z., Lu, W., Li, X., Yang, G., Guo, J., Yu, H., Li, Z., Guan, F.: Altered N-glycan expression profile in epithelial-to-mesenchymal transition of NMuMG cells revealed by an integrated strategy using mass spectrometry and glycogene and lectin microarray analysis. J. Proteome Res. 13(6), 2783–2795 (2014)

    Article  CAS  PubMed  Google Scholar 

  20. Reis, C.A., Osorio, H., Silva, L., Gomes, C., David, L.: Alterations in glycosylation as biomarkers for cancer detection. J. Clin. Pathol. 63(4), 322–329 (2010)

    Article  CAS  PubMed  Google Scholar 

  21. Kam, R.K.T., Poon, T.C.W.: The potentials of Glycomics in biomarker discovery. Clin. Proteomics. 4(3), 67–79 (2008)

    Article  CAS  Google Scholar 

  22. Yu, H., Zhu, M., Qin, Y., Zhong, Y., Yan, H., Wang, Q., Bian, H., Li, Z.: Analysis of glycan-related genes expression and glycan profiles in mice with liver fibrosis. J. Proteome Res. 11(11), 5277–5285 (2012)

    Article  CAS  PubMed  Google Scholar 

  23. Vajaria, B.N., Patel, P.S.: Glycosylation: a hallmark of cancer? Glycoconj J. 34(2), 147–156 (2017)

  24. Qin, Y., Zhong, Y., Ma, T., Wu, F., Wu, H., Yu, H., Huang, C., Li, Z.: Alteration of liver glycopatterns during cirrhosis and tumor progression induced by HBV. Glycoconj. J. 33(2), 125–136 (2016)

    Article  CAS  PubMed  Google Scholar 

  25. Blomme, B., Van Steenkiste, C., Callewaert, N., Van Vlierberghe, H.: Alteration of protein glycosylation in liver diseases. J. Hepatol. 50(3), 592–603 (2009)

    Article  CAS  PubMed  Google Scholar 

  26. Aoyagi, Y., Suzuki, Y., Igarashi, K., Saitoh, A., Oguro, M., Yokota, T., Mori, S., Suda, T., Isemura, M., Asakura, H.: Carbohydrate structures of human alpha-fetoprotein of patients with hepatocellular carcinoma: presence of fucosylated and non-fucosylated triantennary glycans. Br. J. Cancer. 67(3), 486–492 (1993)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Aoyagi, Y., Isokawa, O., Suda, T., Watanabe, M., Suzuki, Y., Asakura, H.: The fucosylation index of alpha-fetoprotein as a possible prognostic indicator for patients with hepatocellular carcinoma. Cancer. 83(10), 2076–2082 (1998)

    Article  CAS  PubMed  Google Scholar 

  28. Vanhooren, V., Liu, X.E., Franceschi, C., Gao, C.F., Libert, C., Contreras, R., Chen, C.: N-glycan profiles as tools in diagnosis of hepatocellular carcinoma and prediction of healthy human ageing. Mech. Ageing Dev. 130(1–2), 92–97 (2009)

    Article  CAS  PubMed  Google Scholar 

  29. Pompach, P., Ashline, D.J., Brnakova, Z., Benicky, J., Sanda, M., Goldman, R.: Protein and site specificity of fucosylation in liver-secreted glycoproteins. J. Proteome Res. 13(12), 5561–5569 (2014)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Kudo, T., Nakagawa, H., Takahashi, M., Hamaguchi, J., Kamiyama, N., Yokoo, H., Nakanishi, K., Nakagawa, T., Kamiyama, T., Deguchi, K., Nishimura, S., Todo, S.: N-glycan alterations are associated with drug resistance in human hepatocellular carcinoma. Mol. Cancer. 6, 32 (2007)

    Article  PubMed  PubMed Central  Google Scholar 

  31. Kamiyama, T., Yokoo, H., Furukawa, J., Kurogochi, M., Togashi, T., Miura, N., Nakanishi, K., Kamachi, H., Kakisaka, T., Tsuruga, Y., Fujiyoshi, M., Taketomi, A., Nishimura, S., Todo, S.: Identification of novel serum biomarkers of hepatocellular carcinoma using glycomic analysis. Hepatology (Baltimore, Md.) 57(6), 2314–2325 (2013)

    Article  CAS  Google Scholar 

  32. Goldman, R., Ressom, H.W., Varghese, R.S., Goldman, L., Bascug, G., Loffredo, C.A., Abdelhamid, M., Gouda, I., Ezzat, S., Kyselova, Z.: Detection of hepatocellular carcinoma using Glycomic analysis. Clin Cancer Res. 15(5), 1808–1813 (2009)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Noda, K., Miyoshi, E., Kitada, T., Nakahara, S., Gao, C.X., Honke, K., Shiratori, Y., Moriwaki, H., Sasaki, Y., Kasahara, A., Hori, M., Hayashi, N., Taniguchi, N.: The enzymatic basis for the conversion of nonfucosylated to fucosylated alpha-fetoprotein by acyclic retinoid treatment in human hepatoma cells: activation of alpha1-6 fucosyltransferase. Tumour Biol. 23(4), 202–211 (2002)

    Article  CAS  PubMed  Google Scholar 

  34. Mehta, A., Block, T.M.: Fucosylated glycoproteins as markers of liver disease. Dis. Markers. 25(4–5), 259–265 (2008)

    Article  CAS  PubMed  Google Scholar 

  35. Comunale, M.A., Rodemichbetesh, L., Hafner, J., Wang, M., Norton, P., Bisceglie, A.M.D., Block, T., Mehta, A.: Linkage specific Fucosylation of alpha-1-antitrypsin in liver Cirrhosis and cancer patients: implications for a biomarker of hepatocellular carcinoma. PLoS One. 5(8), 521–526 (2010)

    Article  Google Scholar 

  36. Comunale, M.A., Lowman, M., Long, R.E., Krakover, J., Philip, R., Seeholzer, S., Evans, A.A., Hann, H.W., Block, T.M., Mehta, A.S.: Proteomic analysis of serum associated fucosylated glycoproteins in the development of primary hepatocellular carcinoma. J. Proteome Res. 5(2), 308–315 (2006)

    Article  CAS  PubMed  Google Scholar 

  37. Aoyagi, Y., Isemura, M., Yosizawa, Z., Suzuki, Y., Sekine, C., Ono, T., Ichida, F.: Fucosylation of serum alpha-fetoprotein in patients with primary hepatocellular carcinoma. Biochim. Biophys. Acta. 830(3), 217–223 (1985)

    Article  CAS  PubMed  Google Scholar 

  38. Vajaria, B.N., Patel, K.R., Begum, R., Patel, P.S.: Sialylation: an avenue to target cancer cells. Pathol Oncol Res : POR. 22(3), 443–447 (2016)

    Article  CAS  PubMed  Google Scholar 

  39. Mehta, A., Herrera, H., Block, T.: Glycosylation and liver cancer. Adv. Cancer Res. 126, 257–279 (2015)

    Article  PubMed  PubMed Central  Google Scholar 

  40. Dall'Olio, F., Chiricolo, M., D'Errico, A., Gruppioni, E., Altimari, A., Fiorentino, M., Grigioni, W.F.: Expression of β-galactoside α2,6 sialyltransferase and of α2,6-sialylated glycoconjugates in normal human liver, hepatocarcinoma, and cirrhosis. Glycobiology. 14(1), 39–49 (2004)

    Article  PubMed  Google Scholar 

  41. Yamashita, K., Tachibana, Y., Nakayama, T., Kitamura, M., Endo, Y., Kobata, A.: Structural studies of the sugar chains of human parotid alpha-amylase. J. Biol. Chem. 255(12), 5635–5642 (1980)

    CAS  PubMed  Google Scholar 

  42. Takashima, S., Amano, J.: Glycosylation and secretion of human α-amylases. Advances in Biological Chemistry. 2(1), 10–19 (2012)

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Acknowledgements

This work is supported by the National Natural Science Foundation (Grant No. 81372365).

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Authors and Affiliations

  1. The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an Jiaotong University Health Science Center, 277 Yanta Xilu, Xi’an, 710061, China

    Yannan Qin & Baozhen Li

  2. Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, No. 229 Taibai Beilu, Xi’an, 710069, China

    Yaogang Zhong, Tianran Ma, Jiaxu Zhang & Zheng Li

  3. The Key Laboratory of Carbohydrate Chemistry & Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China

    Ganglong Yang & Feng Guan

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  1. Yannan Qin
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Correspondence to Zheng Li or Baozhen Li.

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Qin, Y., Zhong, Y., Ma, T. et al. A pilot study of salivary N-glycome in HBV-induced chronic hepatitis, cirrhosis, and hepatocellular carcinoma. Glycoconj J 34, 523–535 (2017). https://doi.org/10.1007/s10719-017-9768-5

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