title: Targeting the GD3 acetylation pathway selectively induces apoptosis in glioblastoma.
abstract: The expression of ganglioside GD3, which plays crucial roles in normal brain development, decreases in adults but is upregulated in neoplastic cells, where it regulates tumor invasion and survival. Normally a buildup of GD3 induces apoptosis, but this does not occur in gliomas due to formation of 9-O-acetyl GD3 by the addition of an acetyl group to the terminal sialic acid of GD3; this renders GD3 unable to induce apoptosis. Using human biopsy-derived glioblastoma cell cultures, we have carried out a series of molecular manipulations targeting GD3 acetylation pathways. Using immunocytochemistry, flow cytometry, western blotting, and transwell assays, we have shown the existence of a critical ratio between GD3 and 9-O-acetyl GD3, which promotes tumor survival. Thus, we have demonstrated for the first time in primary glioblastoma that cleaving the acetyl group restores GD3, resulting in a reduction in tumor cell viability while normal astrocytes remain unaffected. Additionally, we have shown that glioblastoma viability is reduced due to the induction of mitochondrially mediated apoptosis and that this occurs after mitochondrial membrane depolarization. Three methods of cleaving the acetyl group using hemagglutinin esterase were investigated, and we have shown that the baculovirus vector transduces glioma cells as well as normal astroctyes with a relatively high efficacy. A recombinant baculovirus containing hemagglutinin esterase could be developed for the clinic as an adjuvant therapy for glioma.
authors.affiliation: Cellular and Molecular Neuro-oncology Research Group, Institute Biomedical and Biomolecular Sciences, University of Portsmouth, Portsmouth, UK. suzanne.birks@port.ac.uk
authors.forename: Suzanne M
authors.initials: SM
authors.lastname: Birks
authors.forename: John Owusu
authors.initials: JO
authors.lastname: Danquah
authors.forename: Linda
authors.initials: L
authors.lastname: King
authors.forename: Reinhardt
authors.initials: R
authors.lastname: Vlasak
authors.forename: Dariusz C
authors.initials: DC
authors.lastname: Gorecki
authors.forename: Geoffrey J
authors.initials: GJ
authors.lastname: Pilkington
mesh_terms: D000107:Acetylation; D000328:Adult; D017209:Apoptosis; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology*; D002470:Cell Survival; D002478:Cells, Cultured; D002648:Child; D005732:Gangliosides / Q000378:metabolism*; D005909:Glioblastoma / Q000378:metabolism / Q000473:pathology*; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D053078:Membrane Potential, Mitochondrial; D008928:Mitochondria; D010957:Plasmids; D014760:Viral Fusion Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D005732:Gangliosides; D006389:Hemagglutinins, Viral; D014760:Viral Fusion Proteins; C060814:hemagglutinin esterase; C026226:ganglioside, GD3