title: The ST6GalNAc-I sialyltransferase localizes throughout the Golgi and is responsible for the synthesis of the tumor-associated sialyl-Tn O-glycan in human breast cancer.
abstract: The functional properties of glycoproteins are strongly influenced by their profile of glycosylation, and changes in this profile are seen in malignancy. In mucin-type O-linked glycosylation these changes can result in the production of mucins such as MUC1, carrying shorter sialylated O-glycans, and with different site occupancy. Of the tumor-associated sialylated O-glycans, the disaccharide, sialyl-Tn (sialic acid alpha2,6GalNAc), is expressed by 30% of breast carcinomas and is the most tumor-specific. The ST6GalNAc-I glycosyltransferase, which can catalyze the transfer of sialic acid to GalNAc, shows a highly restricted pattern of expression in normal adult tissues, being largely limited to the gastrointestinal tract and absent in mammary gland. In breast carcinomas, however, a complete correlation between the expression of RNA-encoding ST6GalNAc-I and the expression of sialyl-Tn is evident, demonstrating that the expression of sialyl-Tn results from switching on expression of hST6GalNAc-I. Endogenous or exogenous expression of hST6GalNAc-I (but not ST6GalNAc-II) always results in the expression of sialyl-Tn. This ability to override core 1/core 2 pathways of O- linked glycosylation is explained by the localization of ST6GalNAc-I, which is found throughout the Golgi stacks. The development of a Chinese hamster ovary (CHO) cell line expressing MUC1 and ST6GalNAc-I allowed the large scale production of MUC1 carrying 83% sialyl-Tn O-glycans. The presence of ST6GalNAc-I in the CHO cells reduced the number of O-glycosylation sites occupied in MUC1, from an average of 4.3 to 3.8 per tandem repeat. The availability of large quantities of this MUC1 glycoform will allow the evaluation of its efficacy as an immunogen for immunotherapy of MUC1/STn-expressing tumors.
authors.affiliation: Cancer Research-UK Breast Cancer Biology Group, 3rd Floor, Guy's Hospital, London SE1 9RT, United Kingdom.
authors.forename: Robert
authors.initials: R
authors.lastname: Sewell
authors.forename: Malin
authors.initials: M
authors.lastname: Bäckström
authors.forename: Martin
authors.initials: M
authors.lastname: Dalziel
authors.forename: Steven
authors.initials: S
authors.lastname: Gschmeissner
authors.forename: Hasse
authors.initials: H
authors.lastname: Karlsson
authors.forename: Thomas
authors.initials: T
authors.lastname: Noll
authors.forename: Jochem
authors.initials: J
authors.lastname: Gätgens
authors.forename: Henrik
authors.initials: H
authors.lastname: Clausen
authors.forename: Gunnar C
authors.initials: GC
authors.lastname: Hansson
authors.forename: Joy
authors.initials: J
authors.lastname: Burchell
authors.forename: Joyce
authors.initials: J
authors.lastname: Taylor-Papadimitriou
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000737:chemistry; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000737:chemistry; D015152:Blotting, Northern; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000201:enzymology* / Q000473:pathology; D016466:CHO Cells; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D006224:Cricetinae; D005260:Female; D005434:Flow Cytometry; D006031:Glycosylation; D006056:Golgi Apparatus / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D020014:K562 Cells; D042361:Mammary Glands, Human / Q000378:metabolism; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D008956:Models, Chemical; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012799:Sialyltransferases / Q000737:chemistry* / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D014162:Transfection
chemical_list: D000951:Antigens, Neoplasm; D015295:Antigens, Tumor-Associated, Carbohydrate; D011134:Polysaccharides; D011993:Recombinant Fusion Proteins; C055491:sialosyl-Tn antigen; D012799:Sialyltransferases; C038525:CMP-N-acetylneuraminate-alpha-N-acetylgalactosaminide alpha-2,6-sialyltransferase