title: Expression of Cell-Surface Marker ABCB5 Causes Characteristic Modifications of Glucose, Amino Acid and Phospholipid Metabolism in the G3361 Melanoma-Initiating Cell Line.
abstract: We present a pilot study aimed at determining the effects of expression of ATP-binding cassette member B5 (ABCB5), a previously described marker for melanoma-initiating cells, on cellular metabolism. Metabolic profiles for two groups of human G3361 melanoma cells were compared, i.e. wildtype melanoma cells with intact ABCB5 expression (ABCB5-WT) and corresponding melanoma cell variants with inhibited ABCB5 expression, through shRNA-mediated gene knockdown (ABCB5-KD). A comprehensive metabolomic analysis was performed by using proton and phosphorus NMR spectroscopy of cell extracts to examine water-soluble metabolites and lipids. Parametric and non-parametric statistical analysis of absolute and relative metabolite levels yielded significant differences for compounds involved in glucose, amino acid and phospholipid (PL) metabolism. By contrast, energy metabolism was virtually unaffected by ABCB5 expression. The sum of water-soluble metabolites per total protein was 17% higher in ABCB5-WT vs. ABCB5-KD G3361 variants, but no difference was found for the sum of PLs. Enhanced abundance was particularly pronounced for lactate (+ 23%) and alanine (+ 26%), suggesting an increase in glycolysis and potentially glutaminolysis. Increases in PL degradation products, glycerophosphocholine and glycerophosphoethanolamine (+ 85 and 123%, respectively), and redistributions within the PL pool suggested enhanced membrane PL turnover as a consequence of ABCB5 expression. The possibility of glycolysis modulation by an ABCB5-dependent IL1β-mediated mechanism was supported by functional studies employing monoclonal antibody (mAb)-dependent ABCB5 protein inhibition in wildtype G3361 melanoma cells. Our metabolomic results suggest that the underlying biochemical pathways may offer targets for melanoma therapy, potentially in combination with other treatment forms.
authors: Centre de Résonance Magnétique Biologique et Médicale, Unité Mixte de Recherche 7339 Centre National de la Recherche Scientifique, Faculté de Médecine de la Timone, Aix-Marseille Université, Marseille, France.
authors: Norbert W
authors: NW
authors: Lutz
authors: Transplant Research Program, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
authors: Pallavi
authors: P
authors: Banerjee
authors: Transplant Research Program, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.|Harvard Skin Disease Research Center, Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
authors: Brian J
authors: BJ
authors: Wilson
authors: Transplant Research Program, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
authors: Jie
authors: J
authors: Ma
authors: Centre de Résonance Magnétique Biologique et Médicale, Unité Mixte de Recherche 7339 Centre National de la Recherche Scientifique, Faculté de Médecine de la Timone, Aix-Marseille Université, Marseille, France.|Singapore Imaging Consortium, Agency for Science, Technology and Research, Singapore.
authors: Patrick J
authors: PJ
authors: Cozzone
authors: Transplant Research Program, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.|Harvard Skin Disease Research Center, Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.|School of Medical Sciences, Edith Cowan University, Joondalup, Western Australia, Australia.
authors: Markus H
authors: MH
authors: Frank
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000378:metabolism*; D000409:Alanine / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D005947:Glucose / Q000378:metabolism*; D005997:Glycerylphosphorylcholine / Q000378:metabolism; D006801:Humans; D007773:Lactates / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008545:Melanoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D055432:Metabolomics / Q000379:methods; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D010865:Pilot Projects; D034622:RNA Interference
chemical_list: C480744:ABCB5 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000596:Amino Acids; D014408:Biomarkers, Tumor; D007773:Lactates; D010714:Phosphatidylethanolamines; D010743:Phospholipids; C002449:glycerophosphoethanolamine; D005997:Glycerylphosphorylcholine; D005947:Glucose; D000409:Alanine