Characterization of Macrophage Galactose-type Lectin (MGL) Ligands in Colorectal Cancer Cell Lines

Martina Pirro, Yoann Rombouts, Alexandre Stella, Olivier Neyrolles, Odile Burlet-Schiltz, SJ van Vliet, Arnoud de Ru, Yassene Mohammed, Manfred Wuhrer, P.A. van Veelen, P Hensbergen

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Background: The Ca2+-dependent C-type lectin receptor Macrophage Galactose-type Lectin (MGL) is highly expressed by tolerogenic dendritic cells (DC) and macrophages. MGL exhibits a high binding specificity for terminal alpha- and beta-linked GalNAc residues found in Tn, sTn and LacdiNAc antigens. These glycan epitopes are often overexpressed in colorectal cancer (CRC), and, as such, MGL can be used to discriminate tumor from the corresponding healthy tissues. Moreover, the high expression of MGL ligands is associated with poor disease-free survival in stage III of CRC tumors. Nonetheless, the glycoproteins expressed by tumor cells that are recognized by MGL have hitherto remained elusive. Methods: Using a panel of three CRC cell lines (HCT116, HT29 and LS174T), recapitulating CRC diversity, we performed FACS staining and pull-down assays using a recombinant soluble form of MGL (and a mutant MGL as control) combined with mass spectrometry-based (glyco)proteomics. Results: HCT116 and HT29, but not LS174T, are high MGL-binding CRC cell lines. On these cells, the major cell surface binding proteins are receptors (e.g. MET, PTK7, SORL1, PTPRF) and integrins (ITGB1, ITGA3). From these proteins, several N- and/or O-glycopeptides were identified, of which some carried either a LacdiNAc or Tn epitope. Conclusions: We have identified cell surface MGL-ligands on CRC cell lines. General significance: Advances in (glyco)proteomics have led to identification of candidate key mediators of immune-evasion and tumor growth in CRC.

Original languageEnglish
Article number129513
JournalBiochimica et Biophysica Acta (BBA) - General Subjects
Issue number4
Publication statusPublished - Apr 2020


  • C-type lectin
  • Colorectal cancer
  • Glycoproteomics
  • LacdiNAc
  • Tn antigen

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