title: Identification of the determinants of 2-deoxyglucose sensitivity in cancer cells by shRNA library screening.
abstract: Combining glycolytic inhibition with other anti-cancer therapies is a potential approach to treating cancer. In this context, we attempted to identify genes that determine sensitivity to 2-deoxyglucose (2DG), a glycolytic inhibitor, in cancer cells using pooled shRNA libraries targeting ∼15,000 genes. The screen revealed that COPB1 and ARCN1, which are essential in retrograde transport, as determinants of sensitivity to 2DG: silencing of COPB1 or ARCN1 expression sensitized cells to 2DG toxicity. To address the mechanism of potentiation of 2DG toxicity by inhibition of COPI-mediated transport, we focused on the role of lipolysis as an alternate source of energy upon inhibition of glycolysis. In the process of lipolysis, COPI-mediated transport is required for localization to lipid droplets of adipose triglyceride lipase (ATGL), a key enzyme that produces fatty acids from triacylglycerol as a substrate for β-oxidation. The ATGL inhibitor atglistatin potentiated 2DG toxicity, consistent with a model in which a defect in COPI-mediated transport of ATGL to lipid droplets inhibits energy supply, thereby sensitizing cells to glycolytic inhibition. Collectively, our data demonstrated that a defect in COPI-mediated transport or pharmacological inhibition of ATGL potentiates 2DG toxicity in cancer cells, possibly due to a reduction in the energy supply.
authors.affiliation: Chemical Genetics Laboratory, RIKEN, Wako, Saitama, 351-0198, Japan; AMED-CREST, AMED, Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Kobayashi
authors.affiliation: Chemical Genetics Laboratory, RIKEN, Wako, Saitama, 351-0198, Japan; AMED-CREST, AMED, Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan.
authors.forename: Haruna
authors.initials: H
authors.lastname: Nishimura
authors.affiliation: Chemical Genetics Laboratory, RIKEN, Wako, Saitama, 351-0198, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Matsumoto
authors.affiliation: Chemical Genetics Laboratory, RIKEN, Wako, Saitama, 351-0198, Japan; AMED-CREST, AMED, Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan. Electronic address: yoshidam@riken.jp.
authors.forename: Minoru
authors.initials: M
authors.lastname: Yoshida
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D001693:Biological Transport, Active; D045744:Cell Line, Tumor; D020756:Coatomer Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D003847:Deoxyglucose / Q000008:administration & dosage / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000008:administration & dosage / Q000494:pharmacology; D055785:Gene Knockdown Techniques; D015723:Gene Library; D006019:Glycolysis / Q000187:drug effects / Q000235:genetics; D045325:HCT116 Cells; D006367:HeLa Cells; D006801:Humans; D008049:Lipase / Q000037:antagonists & inhibitors / Q000378:metabolism; D050356:Lipid Metabolism / Q000187:drug effects / Q000235:genetics; D009369:Neoplasms / Q000188:drug therapy* / Q000235:genetics* / Q000378:metabolism; D010671:Phenylurea Compounds / Q000008:administration & dosage / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics*; D000217:Acyltransferases
chemical_list: D000970:Antineoplastic Agents; D020756:Coatomer Protein; D003847:Deoxyglucose; D004791:Enzyme Inhibitors; D008049:Lipase; D010671:Phenylurea Compounds; D034741:RNA, Small Interfering; C585749:atglistatin; C494794:PNPLA2 protein, human; D000217:Acyltransferases
keywords: 2-Deoxyglucose; Glycolysis; Lipolysis; Warburg effect; shRNA library screening