title: Aberrantly glycosylated MUC1 is expressed on the surface of breast cancer cells and a target for antibody-dependent cell-mediated cytotoxicity.
abstract: Protein glycosylation often changes during cancer development, resulting in the expression of cancer-associated carbohydrate antigens. In particular mucins such as MUC1 are subject to these changes. We previously identified an immunodominant Tn-MUC1 (GalNAc-α-MUC1) cancer-specific epitope not covered by immunological tolerance in MUC1 humanized mice and man. The objective of this study was to determine if mouse antibodies to this Tn-MUC1 epitope induce antibody-dependent cellular cytotoxicity (ADCC) pivotal for their potential use in cancer immunotherapy. Binding affinity of mAb 5E5 directed to Tn-MUC1 was investigated using BiaCore. The availability of Tn-MUC1 on the surface of breast cancer cells was evaluated by immunohistochemistry, confocal microscopy, and flow cytometry, followed by in vitro assessment of antibody-dependent cellular cytotoxicity by mAb 5E5. Biacore analysis demonstrated high affinity binding (KD = 1.7 nM) of mAb 5E5 to its target, Tn-MUC1. Immunolabelling with mAb 5E5 revealed surface expression of the Tn-MUC1 epitope in breast cancer tissue and cell lines, and mAb 5E5 induced ADCC in two human breast cancer cell lines, MCF7 and T47D. Aberrantly glycosylated MUC1 is expressed on the surface of breast cancer cells and a target for antibody-dependent cell-mediated cytotoxicity suggesting that antibodies targeting glycopeptide epitopes on mucins are strong candidates for cancer-specific immunotherapies.
authors.affiliation: Copenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, Faculty of Health Sciences, University of Copenhagen, Blegdamsvej 3, Copenhagen N, Denmark.
authors.forename: Kirstine
authors.initials: K
authors.lastname: Lavrsen
authors.forename: Caroline B
authors.initials: CB
authors.lastname: Madsen
authors.forename: Morten G
authors.initials: MG
authors.lastname: Rasch
authors.forename: Anders
authors.initials: A
authors.lastname: Woetmann
authors.forename: Niels
authors.initials: N
authors.lastname: Ødum
authors.forename: Ulla
authors.initials: U
authors.lastname: Mandel
authors.forename: Henrik
authors.initials: H
authors.lastname: Clausen
authors.forename: Anders E
authors.initials: AE
authors.lastname: Pedersen
authors.forename: Hans H
authors.initials: HH
authors.lastname: Wandall
mesh_terms: D000116:Acetylgalactosamine / Q000737:chemistry; D000230:Adenocarcinoma / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000915:Antibody Affinity; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology*; D001943:Breast Neoplasms / Q000276:immunology*; D005260:Female; D006031:Glycosylation; D006801:Humans; D061986:MCF-7 Cells; D018396:Mucin-1 / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; C060192:MUC1 protein, human; D018396:Mucin-1; D000116:Acetylgalactosamine