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Molecular targets for therapy

Pharmacological inhibition of fatty-acid oxidation synergistically enhances the effect of l-asparaginase in childhood ALL cells

Leukemia volume 30, pages 209–218 (2016)Cite this article

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Abstract

l-asparaginase (ASNase), a key component in the treatment of childhood acute lymphoblastic leukemia (ALL), hydrolyzes plasma asparagine and glutamine and thereby disturbs metabolic homeostasis of leukemic cells. The efficacy of such therapeutic strategy will depend on the capacity of cancer cells to adapt to the metabolic challenge, which could relate to the activation of compensatory metabolic routes. Therefore, we studied the impact of ASNase on the main metabolic pathways in leukemic cells. Treating leukemic cells with ASNase increased fatty-acid oxidation (FAO) and cell respiration and inhibited glycolysis. FAO, together with the decrease in protein translation and pyrimidine synthesis, was positively regulated through inhibition of the RagB-mTORC1 pathway, whereas the effect on glycolysis was RagB-mTORC1 independent. As FAO has been suggested to have a pro-survival function in leukemic cells, we tested its contribution to cell survival following ASNase treatment. Pharmacological inhibition of FAO significantly increased the sensitivity of ALL cells to ASNase. Moreover, constitutive activation of the mammalian target of rapamycin pathway increased apoptosis in leukemic cells treated with ASNase, but did not increase FAO. Our study uncovers a novel therapeutic option based on the combination of ASNase and FAO inhibitors.

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References

  1. Pui CH, Evans WE . Treatment of acute lymphoblastic leukemia. N Engl J Med 2006; 354: 166–178.

    Article  CAS  PubMed  Google Scholar 

  2. Amylon MD, Shuster J, Pullen J, Berard C, Link MP, Wharam M et al. Intensive high-dose asparaginase consolidation improves survival for pediatric patients with T cell acute lymphoblastic leukemia and advanced stage lymphoblastic lymphoma: a Pediatric Oncology Group study. Leukemia 1999; 13: 335–342.

    Article  CAS  PubMed  Google Scholar 

  3. Moghrabi A, Levy DE, Asselin B, Barr R, Clavell L, Hurwitz C et al. Results of the Dana-Farber Cancer Institute ALL Consortium Protocol 95-01 for children with acute lymphoblastic leukemia. Blood 2007; 109: 896–904.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Jaccard A, Gachard N, Marin B, Rogez S, Audrain M, Suarez F et al. Efficacy of L-asparaginase with methotrexate and dexamethasone (AspaMetDex regimen) in patients with refractory or relapsing extranodal NK/T-cell lymphoma, a phase 2 study. Blood 2011; 117: 1834–1839.

    Article  CAS  PubMed  Google Scholar 

  5. Douer D, Yampolsky H, Cohen LJ, Watkins K, Levine AM, Periclou AP et al. Pharmacodynamics and safety of intravenous pegaspargase during remission induction in adults aged 55 years or younger with newly diagnosed acute lymphoblastic leukemia. Blood 2007; 109: 2744–2750.

    CAS  PubMed  Google Scholar 

  6. Lorenzi PL, Llamas J, Gunsior M, Ozbun L, Reinhold WC, Varma S et al. Asparagine synthetase is a predictive biomarker of L-asparaginase activity in ovarian cancer cell lines. Mol Cancer Ther 2008; 7: 3123–3128.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Panosyan EH, Wang Y, Xia P, Lee WN, Pak Y, Laks DR et al. Asparagine depletion potentiates the cytotoxic effect of chemotherapy against brain tumors. Mol Cancer Res 2014; 12: 694–702.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Purwaha P, Lorenzi PL, Silva LP, Hawke DH, Weinstein JN . Targeted metabolomic analysis of amino acid response to L-asparaginase in adherent cells. Metabolomics 2014; 10: 909–919.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Sircar K, Huang H, Hu L, Cogdell D, Dhillon J, Tzelepi V et al. Integrative molecular profiling reveals asparagine synthetase is a target in castration-resistant prostate cancer. Am J Pathol 2012; 180: 895–903.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Yu M, Henning R, Walker A, Kim G, Perroy A, Alessandro R et al. L-asparaginase inhibits invasive and angiogenic activity and induces autophagy in ovarian cancer. J Cell Mol Med 2012; 16: 2369–2378.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Capizzi RL, Bertino JR, Skeel RT, Creasey WA, Zanes R, Olayon C et al. L-asparaginase: clinical, biochemical, pharmacological, and immunological studies. Ann Intern Med 1971; 74: 893–901.

    Article  CAS  PubMed  Google Scholar 

  12. Cooney DA, Capizzi RL, Handschumacher RE . Evaluation of L-asparagine metabolism in animals and man. Cancer Res 1970; 30: 929–935.

    CAS  PubMed  Google Scholar 

  13. Miller HK, Salser JS, Balis ME . Amino acid levels following L-asparagine amidohydrolase (EC.3.5.1.1) therapy. Cancer Res 1969; 29: 183–187.

    CAS  PubMed  Google Scholar 

  14. Ohnuma T, Holland JF, Freeman A, Sinks LF . Biochemical and pharmacological studies with asparaginase in man. Cancer Res 1970; 30: 2297–2305.

    CAS  PubMed  Google Scholar 

  15. Fine BM, Kaspers GJ, Ho M, Loonen AH, Boxer LM . A genome-wide view of the in vitro response to l-asparaginase in acute lymphoblastic leukemia. Cancer Res 2005; 65: 291–299.

    CAS  PubMed  Google Scholar 

  16. Holleman A, Cheok MH, den Boer ML, Yang W, Veerman AJ, Kazemier KM et al. Gene-expression patterns in drug-resistant acute lymphoblastic leukemia cells and response to treatment. N Engl J Med 2004; 351: 533–542.

    Article  CAS  PubMed  Google Scholar 

  17. Kaspers GJ, Veerman AJ, Pieters R, Van Zantwijk CH, Smets LA, Van Wering ER et al. In vitro cellular drug resistance and prognosis in newly diagnosed childhood acute lymphoblastic leukemia. Blood 1997; 90: 2723–2729.

    CAS  PubMed  Google Scholar 

  18. Krejci O, Starkova J, Otova B, Madzo J, Kalinova M, Hrusak O et al. Upregulation of asparagine synthetase fails to avert cell cycle arrest induced by L-asparaginase in TEL/AML1-positive leukaemic cells. Leukemia 2004; 18: 434–441.

    Article  CAS  PubMed  Google Scholar 

  19. Hermanova I, Zaliova M, Trka J, Starkova J . Low expression of asparagine synthetase in lymphoid blasts precludes its role in sensitivity to L-asparaginase. Exp Hematol 2012; 40: 657–665.

    Article  CAS  PubMed  Google Scholar 

  20. Vander Heiden MG, Cantley LC, Thompson CB . Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 2009; 324: 1029–1033.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Boag JM, Beesley AH, Firth MJ, Freitas JR, Ford J, Hoffmann K et al. Altered glucose metabolism in childhood pre-B acute lymphoblastic leukaemia. Leukemia 2006; 20: 1731–1737.

    Article  CAS  PubMed  Google Scholar 

  22. Scotland S, Saland E, Skuli N, de Toni F, Boutzen H, Micklow E et al. Mitochondrial energetic and AKT status mediate metabolic effects and apoptosis of metformin in human leukemic cells. Leukemia 2013; 27: 2129–2138.

    Article  CAS  PubMed  Google Scholar 

  23. Buzzai M, Jones RG, Amaravadi RK, Lum JJ, DeBerardinis RJ, Zhao F et al. Systemic treatment with the antidiabetic drug metformin selectively impairs p53-deficient tumor cell growth. Cancer Res 2007; 67: 6745–6752.

    Article  CAS  PubMed  Google Scholar 

  24. Hirsch HA, Iliopoulos D, Tsichlis PN, Struhl K . Metformin selectively targets cancer stem cells, and acts together with chemotherapy to block tumor growth and prolong remission. Cancer Res 2009; 69: 7507–7511.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Iiboshi Y, Papst PJ, Hunger SP, Terada N . L-Asparaginase inhibits the rapamycin-targeted signaling pathway. Biochem Biophys Res Commun 1999; 260: 534–539.

    Article  CAS  PubMed  Google Scholar 

  26. Willems L, Jacque N, Jacquel A, Neveux N, Maciel TT, Lambert M et al. Inhibiting glutamine uptake represents an attractive new strategy for treating acute myeloid leukemia. Blood 2013; 122: 3521–3532.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Yang L, Moss T, Mangala LS, Marini J, Zhao H, Wahlig S et al. Metabolic shifts toward glutamine regulate tumor growth, invasion and bioenergetics in ovarian cancer. Mol Syst Biol 2014; 10: 728.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Behrends C, Sowa ME, Gygi SP, Harper JW . Network organization of the human autophagy system. Nature 2010; 466: 68–76.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Hay N, Sonenberg N . Upstream and downstream of mTOR. Genes Dev 2004; 18: 1926–1945.

    Article  CAS  PubMed  Google Scholar 

  30. Holz MK, Ballif BA, Gygi SP, Blenis J . mTOR and S6K1 mediate assembly of the translation preinitiation complex through dynamic protein interchange and ordered phosphorylation events. Cell 2005; 123: 569–580.

    Article  CAS  PubMed  Google Scholar 

  31. Kim E, Goraksha-Hicks P, Li L, Neufeld TP, Guan KL . Regulation of TORC1 by Rag GTPases in nutrient response. Nat Cell Biol 2008; 10: 935–945.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Pieters R, Loonen AH, Huismans DR, Broekema GJ, Dirven MW, Heyenbrok MW et al. In vitro drug sensitivity of cells from children with leukemia using the MTT assay with improved culture conditions. Blood 1990; 76: 2327–2336.

    CAS  PubMed  Google Scholar 

  33. Chou TC . Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev 2006; 58: 621–681.

    Article  CAS  PubMed  Google Scholar 

  34. Deberardinis RJ, Lum JJ, Thompson CB . Phosphatidylinositol 3-kinase-dependent modulation of carnitine palmitoyltransferase 1A expression regulates lipid metabolism during hematopoietic cell growth. J Biol Chem 2006; 281: 37372–37380.

    Article  CAS  PubMed  Google Scholar 

  35. Gerhart-Hines Z, Rodgers JT, Bare O, Lerin C, Kim SH, Mostoslavsky R et al. Metabolic control of muscle mitochondrial function and fatty acid oxidation through SIRT1/PGC-1alpha. EMBO J 2007; 26: 1913–1923.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Ito K, Carracedo A, Weiss D, Arai F, Ala U, Avigan DE et al. A PML-PPAR-delta pathway for fatty acid oxidation regulates hematopoietic stem cell maintenance. Nat Med 2012; 18: 1350–1358.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Sancak Y, Peterson TR, Shaul YD, Lindquist RA, Thoreen CC, Bar-Peled L et al. The Rag GTPases bind raptor and mediate amino acid signaling to mTORC1. Science 2008; 320: 1496–1501.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Alberich-Jorda M, Wouters B, Balastik M, Shapiro-Koss C, Zhang H, Di Ruscio A et al. C/EBPgamma deregulation results in differentiation arrest in acute myeloid leukemia. J Clin Invest 2012; 122: 4490–4504.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Muller HJ, Beier R, Loning L, Blutters-Sawatzki R, Dorffel W, Maass E et al. Pharmacokinetics of native Escherichia coli asparaginase (Asparaginase medac) and hypersensitivity reactions in ALL-BFM 95 reinduction treatment. Br J Haematol 2001; 114: 794–799.

    Article  CAS  PubMed  Google Scholar 

  40. Robitaille AM, Christen S, Shimobayashi M, Cornu M, Fava LL, Moes S et al. Quantitative phosphoproteomics reveal mTORC1 activates de novo pyrimidine synthesis. Science 2013; 339: 1320–1323.

    Article  CAS  PubMed  Google Scholar 

  41. Yamamoto A, Tagawa Y, Yoshimori T, Moriyama Y, Masaki R, Tashiro Y . Bafilomycin A1 prevents maturation of autophagic vacuoles by inhibiting fusion between autophagosomes and lysosomes in rat hepatoma cell line, H-4-II-E cells. Cell Struct Funct 1998; 23: 33–42.

    Article  CAS  PubMed  Google Scholar 

  42. Duran RV, Oppliger W, Robitaille AM, Heiserich L, Skendaj R, Gottlieb E et al. Glutaminolysis activates Rag-mTORC1 signaling. Mol Cell 2012; 47: 349–358.

    Article  CAS  PubMed  Google Scholar 

  43. Samudio I, Harmancey R, Fiegl M, Kantarjian H, Konopleva M, Korchin B et al. Pharmacologic inhibition of fatty acid oxidation sensitizes human leukemia cells to apoptosis induction. J Clin Invest 2010; 120: 142–156.

    Article  CAS  PubMed  Google Scholar 

  44. Estan MC, Calvino E, Calvo S, Guillen-Guio B, Boyano-Adanez Mdel C, de Blas E et al. Apoptotic efficacy of etomoxir in human acute myeloid leukemia cells. Cooperation with arsenic trioxide and glycolytic inhibitors, and regulation by oxidative stress and protein kinase activities. PloS One 2014; 9: e115250.

    Article  PubMed  PubMed Central  Google Scholar 

  45. Vander Heiden MG . Targeting cancer metabolism: a therapeutic window opens. Nat Rev Drug Discov 2011; 10: 671–684.

    Article  CAS  PubMed  Google Scholar 

  46. Lumeng L, Bremer J, Davis EJ . Suppression of the mitochondrial oxidation of (-)-palmitylcarnitine by the malate-aspartate and alpha-glycerophosphate shuttles. J Biol Chem 1976; 251: 277–284.

    CAS  PubMed  Google Scholar 

  47. Schafer ZT, Grassian AR, Song L, Jiang Z, Gerhart-Hines Z, Irie HY et al. Antioxidant and oncogene rescue of metabolic defects caused by loss of matrix attachment. Nature 2009; 461: 109–113.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Carracedo A, Cantley LC, Pandolfi PP . Cancer metabolism: fatty acid oxidation in the limelight. Nat Rev Cancer 2013; 13: 227–232.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Carracedo A, Weiss D, Leliaert AK, Bhasin M, de Boer VC, Laurent G et al. A metabolic prosurvival role for PML in breast cancer. J Clin Invest 2012; 122: 3088–3100.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Jeon SM, Chandel NS, Hay N . AMPK regulates NADPH homeostasis to promote tumour cell survival during energy stress. Nature 2012; 485: 661–665.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Li J, Zhao S, Zhou X, Zhang T, Zhao L, Miao P et al. Inhibition of lipolysis by mercaptoacetate and etomoxir specifically sensitize drug-resistant lung adenocarcinoma cell to paclitaxel. PloS One 2013; 8: e74623.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Tung S, Shi Y, Wong K, Zhu F, Gorczynski R, Laister RC et al. PPARalpha and fatty acid oxidation mediate glucocorticoid resistance in chronic lymphocytic leukemia. Blood 2013; 122: 969–980.

    Article  CAS  PubMed  Google Scholar 

  53. Barger JF, Gallo CA, Tandon P, Liu H, Sullivan A, Grimes HL et al. S6K1 determines the metabolic requirements for BCR-ABL survival. Oncogene 2013; 32: 453–461.

    Article  CAS  PubMed  Google Scholar 

  54. Sengupta S, Peterson TR, Laplante M, Oh S, Sabatini DM . mTORC1 controls fasting-induced ketogenesis and its modulation by ageing. Nature 2010; 468: 1100–1104.

    Article  CAS  PubMed  Google Scholar 

  55. Um SH, Frigerio F, Watanabe M, Picard F, Joaquin M, Sticker M et al. Absence of S6K1 protects against age- and diet-induced obesity while enhancing insulin sensitivity. Nature 2004; 431: 200–205.

    Article  CAS  PubMed  Google Scholar 

  56. Laplante M, Sabatini DM . An emerging role of mTOR in lipid biosynthesis. Curr Biol 2009; 19: R1046–R1052.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Degenhardt K, Mathew R, Beaudoin B, Bray K, Anderson D, Chen G et al. Autophagy promotes tumor cell survival and restricts necrosis, inflammation, and tumorigenesis. Cancer Cell 2006; 10: 51–64.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Galluzzi L, Pietrocola F, Levine B, Kroemer G . Metabolic control of autophagy. Cell 2014; 159: 1263–1276.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Chan WK, Lorenzi PL, Anishkin A, Purwaha P, Rogers DM, Sukharev S et al. The glutaminase activity of L-asparaginase is not required for anticancer activity against ASNS-negative cells. Blood 2014; 123: 3596–3606.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by the Grant Agency of the Ministry of Health of the Czech Republic (NT12429) (IH, JS, JT), (NT12370-5) (TM); The Charles University Grant Agency (632513) (IH, JS, JT), (14-21095P) (KV); UNCE 204025/2012 (KV, PN); Ministry of Health, Czech Republic, (00064203 University Hospital Motol, Prague, Czech Republic) (IH, JS, JT, KF); Czech Science Foundation (P302/12/G01) (MZ); 15-28848A (JS, JT); MSMT Navrat grant LK21307 (MAJ); the Grant Agency of the Czech Republic (14-36804G) (TM); the Ramón y Cajal award, the Basque Department of Industry, Tourism and Trade (Etortek), health (2012111086) and education (PI2012-03), Marie Curie (277043), Movember, ISCIII (PI10/01484, PI13/00031) and ERC (336343) (AC); The Basque Government of education (AAA).

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Authors and Affiliations

  1. Department of Pediatric Hematology/Oncology, CLIP-Childhood Leukaemia Investigation Prague, Charles University Prague, Prague, Czech Republic,

    I Hermanova, K Fiser, M Zaliova, J Trka & J Starkova

  2. CIC bioGUNE Technology Park of Bizkaia, Derio, Spain

    A Arruabarrena-Aristorena, S Fernandez-Ruiz & A Carracedo

  3. Laboratory of Structural Biology and Cell Signaling, Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic

    K Valis, D Kavan & P Novak

  4. Department of Biochemistry, Charles University, Prague, Czech Republic

    K Valis, D Kavan & P Novak

  5. Department of Bioenergetics, Institute of Physiology of the Czech Academy of Sciences, Prague, Czech Republic

    H Nuskova, A Pecinova, J Houstek & T Mracek

  6. Laboratory of Molecular Immunology, Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, Czech Republic

    M Alberich-Jorda

  7. Equipe 11 labellisée par la Ligue Nationale Contre le Cancer,

    M Niso-Santano & G Kroemer

  8. INSERM U1138, Paris, France

    M Niso-Santano & G Kroemer

  9. Centre de Recherche des Cordeliers, Paris, France

    M Niso-Santano & G Kroemer

  10. Metabolomics and Molecular Cell Biology Platforms,

    M Niso-Santano & G Kroemer

  11. Gustave Roussy, Villejuif, France

    M Niso-Santano & G Kroemer

  12. Pôle de Biologie,

    G Kroemer

  13. AP-HP,

    G Kroemer

  14. Hôpital Européen Georges Pompidou, Paris, France

    G Kroemer

  15. Université Paris Descartes,

    G Kroemer

  16. Sorbonne Paris Cité, Paris, France

    G Kroemer

  17. Ikerbasque, Basque Foundation for Science, Bilbao, Spain

    A Carracedo

  18. Department of Biochemistry and Molecular biology, University of the Basque Country, Leioa, Spain

    A Carracedo

  19. University Hospital Motol, Prague, Czech Republic

    J Trka

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  1. I Hermanova
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Correspondence to J Trka.

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Hermanova, I., Arruabarrena-Aristorena, A., Valis, K. et al. Pharmacological inhibition of fatty-acid oxidation synergistically enhances the effect of l-asparaginase in childhood ALL cells. Leukemia 30, 209–218 (2016). https://doi.org/10.1038/leu.2015.213

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