title: Extracellular Vesicles from Ovarian Carcinoma Cells Display Specific Glycosignatures.
abstract: Cells release vesicles to the extracellular environment with characteristic nucleic acid, protein, lipid, and glycan composition. Here we have isolated and characterized extracellular vesicles (EVs) and total cell membranes (MBs) from ovarian carcinoma OVMz cells. EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. Lectin blotting with a panel of lectins showed that EVs had specific glycosignatures relative to MBs. Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. In conclusion, the results showed that glycosignatures of EVs were specific and altered glycosylation within the cell affected the composition and/or dynamics of EVs release. Furthermore, the identified glycosignatures of EVs could provide novel biomarkers for ovarian cancer.
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, Oeiras 2780-157, Portugal. jgomes@itqb.unl.pt.
authors.forename: Joana
authors.initials: J
authors.lastname: Gomes
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, Oeiras 2780-157, Portugal.|BET, Instituto de Biologia Experimental e Tecnológica, Oeiras 2780-157, Portugal.
authors.forename: Patrícia
authors.initials: P
authors.lastname: Gomes-Alves
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, Oeiras 2780-157, Portugal. sofiacarvalho@itqb.unl.pt.|BET, Instituto de Biologia Experimental e Tecnológica, Oeiras 2780-157, Portugal. sofiacarvalho@itqb.unl.pt.
authors.forename: Sofia B
authors.initials: SB
authors.lastname: Carvalho
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, Oeiras 2780-157, Portugal. peixoto@itqb.unl.pt.|BET, Instituto de Biologia Experimental e Tecnológica, Oeiras 2780-157, Portugal. peixoto@itqb.unl.pt.
authors.forename: Cristina
authors.initials: C
authors.lastname: Peixoto
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, Oeiras 2780-157, Portugal. marques@itqb.unl.pt.|BET, Instituto de Biologia Experimental e Tecnológica, Oeiras 2780-157, Portugal. marques@itqb.unl.pt.
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
authors.affiliation: Skin Cancer Unit, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany. p.altevogt@dkfz.de.|Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Ruprecht-Karl University of Heidelberg, Mannheim 68135, Germany. p.altevogt@dkfz.de.
authors.forename: Peter
authors.initials: P
authors.lastname: Altevogt
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, Oeiras 2780-157, Portugal. jcosta@itqb.unl.pt.
authors.forename: Julia
authors.initials: J
authors.lastname: Costa
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D037102:Lectins / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000473:pathology*
chemical_list: D000470:Alkaloids; D037102:Lectins; C065629:kifunensine
keywords: biomarkers; extracellular vesicles; galectin-3-binding protein; glycosignatures; glycosylation; kifunensine; ovarian cancer