Identification of the determinants of 2-deoxyglucose sensitivity in cancer cells by shRNA library screening

• Published in: Biochemical and biophysical research communications Vol. 467; no. 1; pp. 121 - 127
• Main Authors: Kobayashi, Hiroki, Nishimura, Haruna, Matsumoto, Ken, Yoshida, Minoru
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 06.11.2015
• Subjects: 2-Deoxyglucose; Antineoplastic Agents - pharmacology; Biological Transport, Active; Cell Line, Tumor; Coatomer Protein - antagonists & inhibitors; Coatomer Protein - genetics; Coatomer Protein - metabolism; Deoxyglucose - administration & dosage; Deoxyglucose - pharmacology; Drug Resistance, Neoplasm - genetics; Drug Synergism; Enzyme Inhibitors - administration & dosage; Enzyme Inhibitors - pharmacology; Gene Knockdown Techniques; Gene Library; Glycolysis; Glycolysis - drug effects; Glycolysis - genetics; HCT116 Cells; HeLa Cells; Humans; Lipase - antagonists & inhibitors; Lipase - metabolism; Lipid Metabolism - drug effects; Lipid Metabolism - genetics; Lipolysis; Neoplasms - drug therapy; Neoplasms - genetics; Neoplasms - metabolism; Phenylurea Compounds - administration & dosage; Phenylurea Compounds - pharmacology; RNA, Small Interfering - genetics; shRNA library screening; Warburg effect; 2-Deoxyglucose; Glycolysis; shRNA library screening; Lipolysis; Warburg effect
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2015.09.106

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. •We screened for the determinants of 2-deoxyglucose (2DG) sensitivity.•COPB1 and ARCN1 were identified as determinants of 2DG sensitivity.•Inhibition of lipolysis by knockdown of COPB1 or ARCN1 sensitizes cells to 2DG.•Inhibition of lipolysis by atglistatin also sensitizes cells to 2DG.•The combination of 2DG and inhibition of lipolysis could be used to treat cancer.