title: Altered (neo-) lacto series glycolipid biosynthesis impairs α2-6 sialylation on N-glycoproteins in ovarian cancer cells.
abstract: The (neo-) lacto series glycosphingolipids (nsGSLs) comprise of glycan epitopes that are present as blood group antigens, act as primary receptors for human pathogens and are also increasingly associated with malignant diseases. Beta-1, 3-N-acetyl-glucosaminyl-transferase 5 (B3GNT5) is suggested as the key glycosyltransferase for the biosynthesis of nsGSLs. In this study, we investigated the impact of CRISPR-Cas9 -mediated gene disruption of B3GNT5 (∆B3GNT5) on the expression of glycosphingolipids and N-glycoproteins by utilizing immunostaining and glycomics-based PGC-UHPLC-ESI-QTOF-MS/MS profiling. ∆B3GNT5 cells lost nsGSL expression coinciding with reduction of α2-6 sialylation on N-glycoproteins. In contrast, disruption of B4GALNT1, a glycosyltransferase for ganglio series GSLs did not affect α2-6 sialylation on N-glycoproteins. We further profiled all known α2-6 sialyltransferase-encoding genes and showed that the loss of α2-6 sialylation is due to silencing of ST6GAL1 expression in ∆B3GNT5 cells. These results demonstrate that nsGSLs are part of a complex network affecting N-glycosylation in ovarian cancer cells.
authors.affiliation: Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, 4031, Switzerland.|Glyco-oncology, Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, 4031, Switzerland.
authors.forename: Shahidul
authors.initials: S
authors.lastname: Alam
authors.affiliation: Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, 4031, Switzerland.
authors.forename: Merrina
authors.initials: M
authors.lastname: Anugraham
authors.affiliation: Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, 4031, Switzerland.
authors.forename: Yen-Lin
authors.initials: YL
authors.lastname: Huang
authors.affiliation: Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, 4031, Switzerland.
authors.forename: Reto S
authors.initials: RS
authors.lastname: Kohler
authors.affiliation: School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, 4132, Switzerland.
authors.forename: Timm
authors.initials: T
authors.lastname: Hettich
authors.affiliation: Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, 4031, Switzerland.
authors.forename: Katharina
authors.initials: K
authors.lastname: Winkelbach
authors.affiliation: Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, 4031, Switzerland.
authors.forename: Yasmin
authors.initials: Y
authors.lastname: Grether
authors.affiliation: Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, 4031, Switzerland.
authors.forename: Mónica Núñez
authors.initials: MN
authors.lastname: López
authors.affiliation: Shemyakin- Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russian Federation.
authors.forename: Nailia
authors.initials: N
authors.lastname: Khasbiullina
authors.affiliation: Shemyakin- Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russian Federation.
authors.forename: Nicolai V
authors.initials: NV
authors.lastname: Bovin
authors.affiliation: School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, 4132, Switzerland.
authors.forename: Götz
authors.initials: G
authors.lastname: Schlotterbeck
authors.affiliation: Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, 4031, Switzerland.|Glyco-oncology, Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, 4031, Switzerland.
authors.forename: Francis
authors.initials: F
authors.lastname: Jacob
mesh_terms: D064113:CRISPR-Cas Systems; D045744:Cell Line, Tumor; D005260:Female; D055786:Gene Knockout Techniques; D054794:Glycomics; D006023:Glycoproteins / Q000378:metabolism*; D006028:Glycosphingolipids / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D017350:N-Acetylgalactosaminyltransferases / Q000235:genetics*; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism*
chemical_list: D006023:Glycoproteins; D006028:Glycosphingolipids; D017350:N-Acetylgalactosaminyltransferases; C000598680:B4galnt1 protein, human