Lung adenocarcinoma (LAC) is the most common form of lung cancer that increases in non-smokers at younger age. Altered protein glycosylation is one of the hallmarks of malignancy, its role in cancer progression is still poorly understood. In this study, we report mass spectrometric (MS) analysis of N-glycans released from fresh or defrosted tissue specimens from 24 patients with LAC. Comparison of cancerous versus adjacent healthy tissues revealed substantial differences in N-glycan profiles associated with disease. The significant increase in paucimannose and high-mannose glycans with 6-9 mannose residues and decline in the sialylated complex biantenary core fucosylated glycan with composition NeuAcGal2GlcNAc2Man3GlcNAc2Fuc were general features of tumors. In addition, 42 new N-glycan compositions were detected in cancerous tissues. The prominent changes in advanced disease stages were mostly observed in core fucosylated N-glycans with additional fucose (Fuc) residue/s and enhanced branching with non-galactosylated N-acetyl-glucosamine (GlcNAc) units. Both of these monosaccharide types were linked preferably on the 6-antenna. Importantly, as compared with noncancerous tissues, a number of these significant changes were clearly detectable early on in stage I. Application of N-glycan data obtained from tissues was next assessed and validated for evaluation of small sized biopsies obtained via bronchoscopy. In summary, observed alterations and data of newly detected N-glycans expand knowledge about the glycosylation in LAC and may contribute to research in more tailored therapies. Moreover, the results demonstrate effectiveness of the presented approach for utility in rapid discrimination of cancerous from healthy lung tissues.

Central European Institute for Technology Masaryk University Kamenice 5 Brno Czech Republic

Department of Pathology University Hospital and Medical Faculty of Masaryk University Brno Czech Republic

Department of Respiratory Diseases and TB University Hospital Brno and Medical Faculty of Masaryk University Brno Czech Republic

National Centre for Biomolecular Research Faculty of Science Masaryk University Brno Czech Republic

RECETOX Faculty of Science Masaryk University Brno Czech Republic

The Institute of Human Virology University of Maryland 725W Lombard St Baltimore MD 21201 USA

See more in PubMed

Torre LA, Siegel RL, Jemal A. Lung cancer statistics. In: Ahmad A, Gadgeel S, editors. Lung cancer and personalized medicine: current knowledge and therapies. Cham: Springer International Publishing, 2016. p. 1–19.

Hashimoto T, Tokuchi Y, Hayashi M, et al. Different subtypes of human lung adenocarcinoma caused by different etiological factors. Am J Pathol. 2000;157:2133–41. DOI

Okayama H, Kohno T, Ishii Y, et al. Identification of genes upregulated in ALK-positive and EGFR/KRAS/ALK-negative lung adenocarcinomas. Cancer Res. 2012;72:100–11. DOI

Cibulskis K, Lawrence MS, Carter SL, et al. Sensitive detection of somatic point mutations in impure and heterogeneous cancer samples. Nat Biotechnol. 2013;31:213–9. DOI

Balani S, Nguyen LV, Eaves CJ. Modeling the process of human tumorigenesis. Nat Commun. 2017;8:15422. DOI

Robles AI, Harris CC. Integration of multiple “OMIC” biomarkers: a precision medicine strategy for lung cancer. Lung Cancer. 2017;107:50–58. DOI

Planck M, Edlund K, Botling J, et al. Genomic and transcriptional alterations in lung adenocarcinoma in relation to EGFR and KRAS mutation status. Plos ONE. 2013;8:e78614. DOI

Network TCGAR. Comprehensive molecular profiling of lung adenocarcinoma. Nature. 2014;511:543–50. DOI

Doria-Rose VP, White MC, Klabunde CN, et al. Use of lung cancer screening tests in the United States: results from the 2010 National Health Interview Survey. Cancer Epidemiol Biomark Prev. 2012;21:1049–59. DOI

Team TNLSTR. Reduced lung-cancer mortality with low-dose computed tomographic screening. N Engl J Med. 2011;365:395. DOI

Chansky K, Sculier J-P, Crowley JJ, et al. The International Association for the Study of Lung Cancer Staging Project: prognostic factors and pathologic TNM stage in surgically managed non-small cell lung cancer. J Thorac Oncol. 2009;4:792–801. DOI

Chen Z, Fillmore CM, Hammerman PS, et al. Non-small-cell lung cancers: a heterogeneous set of diseases. Nat Rev Cancer. 2014;14:535–46. DOI

Dwek RA. Glycobiology: toward understanding the function of sugars. Chem Rev. 1996;96:683–720. DOI

Bertozzi CR, Kiessling LL. Chemical glycobiology. Science. 2001;291:2357–64. DOI

Varki A, Cummings RD, Esko JD, et al., editors. Essentials of glycobiology, 3rd ed. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press, 2015. http://www.ncbi.nlm.nih.gov/books/NBK310274/ . Accessed 29 August 2018.

Taniguchi N, Kizuka Y. Glycans and cancer: role of N-glycans in cancer biomarker, progression and metastasis, and therapeutics. Adv Cancer Res. 2015;126:11–51.

Esposito M, Mondal N, Greco TM, et al. Bone vascular niche E-selectin induces mesenchymal–epithelial transition and Wnt activation in cancer cells to promote bone metastasis. Nat Cell Biol. 2019;21:627–39. DOI

Lattová E, Tomanek B, Bartusik D, et al. N-glycomic changes in human breast carcinoma MCF-7 and T-lymphoblastoid cells after treatment with herceptin and herceptin/lipoplex. J Proteome Res. 2010;9:1533–40. DOI

Mann BF, Goetz JA, House MG, et al. Glycomic and proteomic profiling of pancreatic cyst fluids identifies hyperfucosylated lactosamines on the n-linked glycans of overexpressed glycoproteins. Mol Cell Proteom. 2012;11:M111.015792. DOI

Ji IJ, Hua S, Shin DH, et al. Spatially-resolved exploration of the mouse brain glycome by tissue glyco-capture (TGC) and nano-LC/MS. Anal Chem. 2015;87:2869–77. DOI

Sethi MK, Thaysen-Andersen M, Smith JT, et al. Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting glcnac and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. J Proteome Res. 2014;13:277–88. DOI

Dahmen A-C, Fergen M-T, Laurini C, et al. Paucimannosidic glycoepitopes are functionally involved in proliferation of neural progenitor cells in the subventricular zone. Glycobiology. 2015;25:869–80. DOI

Möginger U, Grunewald S, Hennig R, et al. Alterations of the human skin N- and O-glycome in basal cell carcinoma and squamous cell carcinoma. Front Oncol. 2018;8. https://doi.org/10.3389/fonc.2018.00070 .

Arnold JN, Saldova R, Galligan MC, et al. Novel glycan biomarkers for the detection of lung cancer. J Proteome Res. 2011;10:1755–64. DOI

Vasseur JA, Goetz JA, Alley WR, et al. Smoking and lung cancer-induced changes in N-glycosylation of blood serum proteins. Glycobiology. 2012;22:1684–708. DOI

Pompach P, Ashline DJ, Brnakova Z, et al. Protein and site specificity of fucosylation in liver-secreted glycoproteins. J Proteome Res. 2014;13:5561–9. DOI

Potapenko IO, Haakensen VD, Lüders T, et al. Glycan gene expression signatures in normal and malignant breast tissue; possible role in diagnosis and progression. Mol Oncol. 2010;4:98–118. DOI

Satomaa T, Heiskanen A, Leonardsson I, et al. Analysis of the human cancer glycome identifies a novel group of tumor-associated N-acetylglucosamine glycan antigens. Cancer Res. 2009;69:5811–9. DOI

Ruhaak LR, Taylor SL, Stroble C, et al. Differential N-glycosylation patterns in lung adenocarcinoma tissue. J Proteome Res. 2015;14:4538–49. DOI

Wang X, Deng Z, Huang C, et al. Differential N-glycan patterns identified in lung adenocarcinoma by N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections. J Proteom. 2018;172:1–10. DOI

Lattová E, Bryant J, Skřičková J, et al. Efficient procedure for N-glycan analyses and detection of endo H-like activity in human tumor specimens. J Proteome Res. 2016;15:2777–86. DOI

Ciucanu I, Kerek F. A simple and rapid method for the permethylation of carbohydrates. Carbohydr Res. 1984;131:209–17. DOI

Lattová E, Skřičková J, Zdráhal Z. Applicability of phenylhydrazine labeling for structural studies of fucosylated N-glycans. Anal Chem. 2019;91:7985–90. DOI

Lattová E, Perreault H. The usefulness of hydrazine derivatives for mass spectrometric analysis of carbohydrates. Mass Spectrom Rev. 2013;32:366–85. DOI

Domon B, Costello C. A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates. Glycoconj J. 1988;5:397–409. DOI

Walther T, Karamanska R, Chan RWY, et al. Glycomic analysis of human respiratory tract tissues and correlation with influenza virus infection. PLoS Pathog. 2013;9:e1003223. DOI

Tu H-C, Hsiao Y-C, Yang W-Y, et al. Up-regulation of golgi α-mannosidase IA and down-regulation of golgi α-mannosidase IC activates unfolded protein response during hepatocarcinogenesis. Hepatol Commun. 2017;1:230–47. DOI

Voynow JA, Kaiser RS, Scanlin TF, et al. Purification and characterization of GDP-L-fucose-N-acetyl beta-D-glucosaminide alpha 1–6fucosyltransferase from cultured human skin fibroblasts. Requirement of a specific biantennary oligosaccharide as substrate. J Biol Chem. 1991;266:21572–7. PubMed

Ihara H, Hanashima S, Okada T, et al. Fucosylation of chitooligosaccharides by human α1,6-fucosyltransferase requires a nonreducing terminal chitotriose unit as a minimal structure. Glycobiology. 2010;20:1021–33. DOI

Chen C-Y, Jan Y-H, Juan Y-H, et al. Fucosyltransferase 8 as a functional regulator of nonsmall cell lung cancer. PNAS. 2013;110:630–5. DOI

Lau KS, Dennis JW. N-glycans in cancer progression. Glycobiology. 2008;18:750–60. DOI

N-glycan profiling of tissue samples to aid breast cancer subtyping

Comprehensive N-glycosylation mapping of envelope glycoprotein from tick-borne encephalitis virus grown in human and tick cells