Advertisement

Cellular and Molecular Neurobiology

, Volume 39, Issue 3, pp 415–434 | Cite as

Mammalian Target of Rapamycin 2 (MTOR2) and C-MYC Modulate Glucosamine-6-Phosphate Synthesis in Glioblastoma (GBM) Cells Through Glutamine: Fructose-6-Phosphate Aminotransferase 1 (GFAT1)

  • Bo LiuEmail author
  • Ze-Bin Huang
  • Xin Chen
  • Yi-Xiang See
  • Zi-Kai Chen
  • Huan-Kai Yao
Original Research

Abstract

Glucose and glutamine are two essential ingredients for cell growth. Glycolysis and glutaminolysis can be linked by glutamine: fructose-6-phosphate aminotransferase (GFAT, composed of GFAT1 and GFAT2) that catalyzes the synthesis of glucosamine-6-phosphate and glutamate by using fructose-6-phosphate and glutamine as substrates. The role of mammalian target of rapamycin (MTOR, composed of MTOR1 and MTOR2) in regulating glycolysis has been explored in human cancer cells. However, whether MTOR can interact with GFAT to regulate glucosamine-6-phosphate is poorly understood. In this study, we report that GFAT1 is essential to maintain the malignant features of GBM cells. And MTOR2 rather than MTOR1 plays a robust role in promoting GFAT1 protein activity, and accelerating the progression of glucosamine-6-phosphate synthesis, which is not controlled by the PI3K/AKT signaling. Intriguingly, high level of glucose or glutamine supply promotes MTOR2 protein activity. In turn, up-regulating glycolytic and glutaminolytic metabolisms block MTOR dimerization, enhancing the release of MTOR2 from the MTOR complex. As a transcriptional factor, C-MYC, directly targeted by MTOR2, promotes the relative mRNA expression level of GFAT1. Notably, our data reveal that GFAT1 immunoreactivity is positively correlated with the malignant grades of glioma patients. Kaplan–Meier assay reveals the correlations between patients’ 5-year survival and high GFAT1 protein expression. Taken together, we propose that the MTOR2/C-MYC/GFAT1 axis is responsible for the modulation on the crosstalk between glycolysis and glutaminolysis in GBM cells. Under the condition of accelerated glycolytic and/or glutaminolytic metabolisms, the MTOR2/C-MYC/GFAT1 axis will be up-regulated in GBM cells.

Keywords

Glycolysis Glutaminolysis MTOR2 GFAT1 GBM 

Notes

Acknowledgements

We thank Dr. Qi Zhang (Tiantan Hospital, Beijing, China) for kindly collecting glioma tissue samples and clinical information. This research was funded by the Postdoctoral Research Foundation (2014) provided by the University of Macau, SAR, China.

Author Contributions

B.L. designed the research project, performed most of the experiments, analyzed the data, and wrote the manuscript. Z.B.H, X.C., Y.X.S., Z.K.C., and H.K.Y. performed the experiments and analyzed the data.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical Approval

This study used human glioma tissues and brain injury tissues obtained from Tiantan Hospital (Beijing, China). All procedures performed in studies involving human participants were in accordance with the Ethical Standards of the Institutional and/or National Research Committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

References

  1. Altman BJ, Stine ZE, Dang CV (2016) From Krebs to clinic: glutamine metabolism to cancer therapy. Nat Rev Cancer 16:619–634CrossRefGoogle Scholar
  2. Biggs III WH, Meisenhelder J, Hunter T, Cavenee WK, Arden KC (1999) Protein kinase B/Akt-mediated phosphorylation promotes nuclear exclusion of the winged helix transcription factor FKHR1. Proc Natl Acad Sci USA 96:7421–7426CrossRefGoogle Scholar
  3. Cairns RA, Harris I, McCracken S, Mak TW (2011a) Cancer cell metabolism. Cold Spring Harb Symp Quant Biol 76:299–311CrossRefGoogle Scholar
  4. Cairns RA, Harris IS, Mak TW (2011b) Regulation of cancer cell metabolism. Nat Rev Cancer 11:85–95CrossRefGoogle Scholar
  5. Cantor JR, Sabatini DM (2012) Cancer cell metabolism: one hallmark, many faces. Cancer Discov 2:881–898CrossRefGoogle Scholar
  6. Choe G, Horvath S, Cloughesy TF, Crosby K, Seligson D, Palotie A, Inge L, Smith BL, Sawyers CL, Mischel PS (2003) Analysis of the phosphatidylinositol 30-kinase signaling pathway in GBM patients in vivo. Cancer Res 63:2742–2746Google Scholar
  7. Christofk HR, Vander Heiden MG, Wu N, Asara JM, Cantley LC (2008) Pyruvate kinase M2 is a phosphotyrosine-binding protein. Nature 452:181–186CrossRefGoogle Scholar
  8. Dang CV (2012) Links between metabolism and cancer. Genes Dev 26:877–890CrossRefGoogle Scholar
  9. Dang CV, Le A, Gao P (2009) MYC-induced cancer cell energy metabolism and therapeutic opportunities. Clin Cancer Res 15:6479–6483CrossRefGoogle Scholar
  10. DeBerardinis RJ, Cheng T (2010) Q’s next: the diverse functions of glutamine in metabolism, cell biology and cancer. Oncogene 29:313–324CrossRefGoogle Scholar
  11. DeBerardinis RJ, Mancuso A, Daikhin E, Nissim I, Yudkoff M, Wehrli S et al (2007) Beyond aerobic glycolysis: transformed cells can engage in glutamine metabolism that exceeds the requirement for protein and nucleotide synthesis. Proc Natl Acad Sci USA 104:19345–19350CrossRefGoogle Scholar
  12. DeBerardinis RJ, Lum JJ, Hatzivassiliou G, Thompson CB (2008) The biology of cancer: metabolic reprogramming fuels cell growth and proliferation. Cell Metab 7:11–20CrossRefGoogle Scholar
  13. DeHaven JE, Robinson KA, Nelson BA, Buse MG (2001) A novel variant of glutamine: fructose-6-phosphate amidotransferase-1 (GFAT1) mRNA is selectively expressed in striated muscle. Diabetes 50:2419–2424CrossRefGoogle Scholar
  14. Faubert B, Boily G, Izreig S, Griss T, Samborska B, Dong Z, Dupuy F, Chambers C, Fuerth BJ, Viollet B et al (2013) AMPK is a negative regulator of the Warburg effect and suppresses tumor growth in vivo. Cell Metab 17:113–124CrossRefGoogle Scholar
  15. Gan B, Lim C, Chu G, Hua S, Ding Z, Collins M, Hu J, Jiang S, Fletcher-Sananikone E, Zhuang L et al (2010) FoxOs enforce a progression checkpoint to constrain MTORC1-activated renal tumorigenesis. Cancer Cell 18:472–484CrossRefGoogle Scholar
  16. Gao P, Tchernyshyov I, Chang TC et al (2009) C-MYC suppression of miR23a/b enhances mitochondrial glutaminase expression and glutamine metabolism. Nature 458:762–765CrossRefGoogle Scholar
  17. Guertin DA, Stevens DM, Thoreen CC, Burds AA, Kalaany NY, Moffat J, Brown M, Fitzgerald KJ, Sabatini DM (2006) Ablation in mice of the MTORC components raptor, rictor, or mLST8 reveals that MTORC2 is required for signaling to Akt-FOXO and PKCalpha, but not S6K1. Dev Cell 11:859–871CrossRefGoogle Scholar
  18. Hagiwara A, Cornu M, Cybulski N, Polak P, Betz C, Trapani F, Terracciano L, Heim MH, Rüegg MA, Hall MN (2012) Hepatic MTORC2 activates glycolysis and lipogenesis through Akt, glucokinase, and SREBP1c. Cell Metab 15:725–738CrossRefGoogle Scholar
  19. Hosios AM, Hecht VC, Danai LV, Johnson MO, Rathmell JC, Steinhauser ML et al (2016) Amino acids rather than glucose account for the majority of cell mass in proliferating mammalian cells. Dev Cell 36:540–549CrossRefGoogle Scholar
  20. Hu Y, Riesland L, Paterson AJ, Kudlow JE (2004) Phosphorylation of mouse glutamine-fructose-6-phosphate amidotransferase 2 (GFAT2) by cAMP-dependent protein kinase increases the enzyme activity. J Biol Chem 279:29988–29993CrossRefGoogle Scholar
  21. Hudson CC, Liu M, Chiang GG, Otterness DM, Loomis DC, Kaper F, Giaccia AJ, Abraham RT (2002) Regulation of hypoxia-inducible factor 1alpha expression and function by the mammalian target of rapamycin. Mol Cell Biol 22:7004–7014CrossRefGoogle Scholar
  22. Kaelin WG Jr, Ratcliffe PJ (2008) Oxygen sensing by metazoans: the central role of the HIF hydroxylase pathway. Mol Cell 30:393–402CrossRefGoogle Scholar
  23. Kleszcz R, Paluszczak J, Krajka-Kuźniak V, Baer-Dubowska W (2018) The inhibition of C-MYC transcription factor modulates the expression of glycolytic and glutaminolytic enzymes in FaDu hypopharyngeal carcinoma cells. Adv Clin Exp Med 27:735–742CrossRefGoogle Scholar
  24. Koppenol WH, Bounds PL, Dang CV (2011) Otto Warburg’s contributions to current concepts of cancer metabolism. Nat Rev Cancer 11:325–337CrossRefGoogle Scholar
  25. Le A, Lane AN, Hamaker M, Bose S, Gouw A, Barbi J et al (2012) Glucose-independent glutamine metabolism via TCA cycling for proliferation and survival in B cells. Cell Metab 15:110–121CrossRefGoogle Scholar
  26. Lee M, Theodoropoulou M, Graw J, Roncaroli F, Zatelli MC, Pellegata NS (2011) Levels of p27 sensitize to dual PI3K/mTOR inhibition. Mol Cancer Ther 10:1450–1459CrossRefGoogle Scholar
  27. Levine AJ, Puzio-Kuter AM (2010) The control of the metabolic switch in cancers by oncogenes and tumor suppressor genes. Science 330:134001344CrossRefGoogle Scholar
  28. Lunt SY, Vander Heiden MG (2011) Aerobic glycolysis: meeting the metabolic requirements of cell proliferation. Annu Rev Cell Dev Biol 27:441–464CrossRefGoogle Scholar
  29. Marroquin LD, Hynes J, Dykens JA, Jamieson JD, Will Y (2007) Circumventing the Crabtree effect: replacing media glucose with galactose increases susceptibility of HepG2 cells to mitochondrial toxicants. Toxicol Sci 97:539–547CrossRefGoogle Scholar
  30. Matsui Y, Lai ZC (2019) Bimolecular fluorescence complementation (BiFC) in tissue culture and in developing tissues of Drosophila to study protein–protein interactions. Methods Mol Biol 1893:75–85CrossRefGoogle Scholar
  31. Medina MA, Núñez de Castro I (1990) Glutaminolysis and glycolysis interactions in proliferant cells. Int J Biochem 22:681–683CrossRefGoogle Scholar
  32. Muellner MK, Uras IZ, Gapp BV, Kerzendorfer C, Smida M, Lechtermann H, Craig-Mueller N, Colinge J, Duernberger G, Nijman SM (2011) A chemical-genetic screen reveals a mechanism of resistance to PI3K inhibitors in cancer. Nat Chem Biol 7:787–793CrossRefGoogle Scholar
  33. Niimi M, Ogawara T, Yamashita T, Yamamoto Y, Ueyama A, Kambe T et al (2001) Identification of GFAT1-L, a novel splice variant of human glutamine: fructose-6-phosphate amidotransferase (GFAT1) that is expressed abundantly in skeletal muscle. J Hum Genet 46:566–571CrossRefGoogle Scholar
  34. Oki T, Yamazaki K, Kuromitsu J, Okada M, Tanaka I (1999) cDNA cloning and mapping of a novel subtype of glutamine:fructose-6-phosphate amidotransferase (GFAT2) in human and mouse. Genomics 57:227–234CrossRefGoogle Scholar
  35. Pelicano H, Martin DS, Xu RH, Huang P (2006) Glycolysis inhibition for anticancer treatment. Oncogene 25:4633–4646CrossRefGoogle Scholar
  36. Plas DR, Thompson CB (2005) Akt-dependent transformation: there is more to growth than just surviving. Oncogene 24:7435–7442CrossRefGoogle Scholar
  37. Qie S, Liang D, Yin C, Gu W, Meng M, Wang C et al (2012) Glutamine depletion and glucose depletion trigger growth inhibition via distinctive gene expression reprogramming. Cell Cycle 11:3679–3690CrossRefGoogle Scholar
  38. Rohle D, Popovici-Muller J, Palaskas N, Turcan S, Grommes C, Campos C, Tsoi J, Clark O, Oldrini B, Komisopoulou E et al (2013) An inhibitor of mutant IDH1 delays growth and promotes differentiation of glioma cells. Science 340:626–630CrossRefGoogle Scholar
  39. Shanware NP, Mullen AR, DeBerardinis RJ, Abraham RT (2011) Glutamine: pleiotropic roles in tumor growth and stress resistance. J Mol Med (Berl) 89:229–236CrossRefGoogle Scholar
  40. Tanaka K, Babic I, Nathanson D, Akhavan D, Guo D, Gini B, Dang J, Zhu S, Yang H, De Jesus J et al (2011) Oncogenic EGFR signaling activates an MTORC2-NF-kB pathway that promotes chemotherapy resistance. Cancer Discov 1:524–538CrossRefGoogle Scholar
  41. Tong X, Zhao F, Thompson CB (2009) The molecular determinants of de novo nucleotide biosynthesis in cancer cells. Curr Opin Genet Dev 19:32–37CrossRefGoogle Scholar
  42. Vadla R, Haldar D (2018) Mammalian target of rapamycin complex 2 (MTORC2) controls glycolytic gene expression by regulating Histone H3 Lysine 56 acetylation. Cell Cycle 17:110–123CrossRefGoogle Scholar
  43. Vander Heiden MG, Cantley LC, Thompson CB (2009) Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324:1029–1033CrossRefGoogle Scholar
  44. Vyas B, Silakari O, Bahia MS, Singh B (2013) Glutamine: fructose-6-phosphate amidotransferase (GFAT): homology modelling and designing of new inhibitors using pharmacophore and docking based hierarchical virtual screening protocol. SAR QSAR Environ Res 24:733–752CrossRefGoogle Scholar
  45. Wang B, Moya N, Niessen S, Hoover H, Mihaylova MM, Shaw RJ, Yates JR III, Fischer WH, Thomas JB, Montminy M (2011) A hormone-dependent module regulating energy balance. Cell 145:596–606CrossRefGoogle Scholar
  46. Wang QS, Kong PZ, Li XQ, Yang F, Feng YM (2015) FOXF2 deficiency promotes epithelial-mesenchymal transition and metastasis of basal-like breast cancer. Breast Cancer Res 17:30CrossRefGoogle Scholar
  47. Warburg O (1956) On the origin of cancer cells. Science 123:309–314CrossRefGoogle Scholar
  48. Ward PS, Thompson CB (2012) Metabolic reprogramming: a cancer hallmark even Warburg did not anticipate. Cancer Cell 21:297–308CrossRefGoogle Scholar
  49. Wellen KE, Lu C, Mancuso A, Lemons JM, Ryczko M, Dennis JW et al (2010) The hexosamine biosynthetic pathway couples growth factor-induced glutamine uptake to glucose metabolism. Genes Dev 24:2784–2799CrossRefGoogle Scholar
  50. Wise DR, DeBerardinis RJ, Mancuso A et al (2008) Myc regulates a transcriptional program that stimulates mitochondrial glutaminolysis and leads to glutamine addiction. Proc Natl Acad Sci USA 105:18782–18787CrossRefGoogle Scholar
  51. Wise DR, Thompson CB (2010) Glutamine addiction: a new therapeutic target in cancer. Trends Biochem Sci 35:427–433CrossRefGoogle Scholar
  52. Yang C, Peng P, Li L, Shao M, Zhao J, Wang L, Duan F, Song S, Wu H, Zhang J, Zhao R, Jia D, Zhang M, Wu W, Li C, Rong Y, Zhang L, Ruan Y, Gu J (2016) High expression of GFAT1 predicts poor prognosis in patients with pancreatic cancer. Sci Rep 6:39044CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Laboratory of Molecular GeneticsUniversity of Maryland School of MedicineBaltimoreUSA
  2. 2.Department of Otorhinolaryngology Head & Neck SurgeryUniversity of Maryland School of MedicineBaltimoreUSA
  3. 3.Center of Reproduction, development and aging, Institute of Translational Medicine, Cancer Centre, Faculty of Health SciencesUniversity of MacauHengqinChina

Personalised recommendations