Reduced phosphocholine and hyperpolarized lactate provide magnetic resonance biomarkers of PI3K/Akt/mTOR inhibition in glioblastoma (original) (raw)

Abstract

The phosphatidylinositol-3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway is activated in more than 88% of glioblastomas (GBM). New drugs targeting this pathway are currently in clinical trials. However, noninvasive assessment of treatment response remains challenging. By using magnetic resonance spectroscopy (MRS), PI3K/Akt/ mTOR pathway inhibition was monitored in 3 GBM cell lines (GS-2, GBM8, and GBM6; each with a distinct pathway activating mutation) through the measurement of 2 mechanistically linked MR biomarkers: phosphocholine (PC) and hyperpolarized lactate. 31 P MRS studies showed that treatment with the PI3K inhibitor LY294002 induced significant decreases in PC to 34 %+ + + + + 9% of control in GS-2 cells, 48% + + + + + 5% in GBM8, and 45% + + + + + 4% in GBM6. The mTOR inhibitor everolimus also induced a significant decrease in PC to 62% + + + + + 14%, 57% + + + + + 1%, and 58% + + + + + 1% in GS-2, GBM8, and GBM6 cells, respectively. Using hyperpolarized 13 C MRS, we demonstrated that hyperpolarized lactate levels were significantly decreased following PI3K/Akt/mTOR pathway inhibition in all 3 cell lines to 51% + + + + + 10%, 62% + + + + + 3%, and 58% + + + + + 2% of control with LY294002 and 72% + + + + + 3%, 61% + + + + + 2%, and 66% + + + + + 3% of control with everolimus in GS-2, GBM8, and GBM6 cells, respectively. These effects were mediated by decreases in the activity and expression of choline kinase a and lactate dehydrogenase, which respectively control PC and lactate production downstream of HIF-1. Treatment with the DNA damaging agent temozolomide did not have an effect on either biomarker in any cell line. This study highlights the potential of PC and hyperpolarized lactate as noninvasive MR biomarkers of response to targeted inhibitors in GBM.

Loading...

Loading Preview

Sorry, preview is currently unavailable. You can download the paper by clicking the button above.

References (52)

  1. CBRTUS. Statistical report -Primary brain and central nervous system tumors diagnosed in the United states in 2004 -2006. 2010; http:// seer.cancer.gov/.
  2. Clarke J, Butowski N, Chang S. Recent advances in therapy for glioblast- oma. Arch Neurol. 2010;67(3):279 -283.
  3. Furnari FB, Fenton T, Bachoo RM, et al. Malignant astrocytic glioma: genet- ics, biology, and paths to treatment. Genes Dev. 2007;21(21):2683-2710.
  4. Grzmil M, Hemmings BA. Deregulated signalling networks in human brain tumours. Biochim Biophys Acta. 2010;1804(3):476 -483.
  5. Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature. 2008;455(7216):1061 -1068.
  6. Billottet C, Grandage VL, Gale RE, et al. A selective inhibitor of the p110delta isoform of PI 3-kinase inhibits AML cell proliferation and sur- vival and increases the cytotoxic effects of VP16. Oncogene. 2006;25(50):6648 -6659.
  7. Garlich JR, De P, Dey N, et al. A vascular targeted pan phosphoinositide 3-kinase inhibitor prodrug, SF1126, with antitumor and antiangiogenic activity. Cancer Res. 2008;68(1): 206 -215.
  8. Negendank W. Studies of human tumors by MRS: a review. NMR Biomed. 1992;5(5):303 -324.
  9. Kvistad KA, Bakken IJ, Gribbestad IS, et al. Characterization of neoplas- tic and normal human breast tissues with in vivo (1)H MR spectroscopy. J Magn Reson Imaging. 1999;10(2):159 -164.
  10. de Certaines JD, Larsen VA, Podo F, Carpinelli G, Briot O, Henriksen O. In vivo 31P MRS of experimental tumours. NMR Biomed. 1993;6(6):345 -365.
  11. Evelhoch JL, Gillies RJ, Karczmar GS, et al. Applications of magnetic res- onance in model systems: cancer therapeutics. Neoplasia. 2000;2(1 - 2):152 -165.
  12. Venkatesh et al.: PC and lactate -biomarkers of PI3K inhibition in GBM NEURO-ONCOLOGY † M A R C H 2 0 1 2 323 at University of California, San Francisco on February 6, 2013 http://neuro-oncology.oxfordjournals.org/ Downloaded from
  13. Leach MO, Verrill M, Glaholm J, et al. Measurements of human breast cancer using magnetic resonance spectroscopy: a review of clinical measurements and a report of localized 31P mea- surements of response to treatment. NMR Biomed. 1998;11(7):314 -340.
  14. Vigneron D, Bollen A, McDermott M, et al. Three-dimensional magnet- ic resonance spectroscopic imaging of histologically confirmed brain tumors. Magn Reson Imaging. 2001;19(1):89 -101.
  15. Koul D, Shen R, Kim YW, et al. Cellular and in vivo activity of a novel PI3K inhibitor, PX-866, against human glioblastoma. Neuro Oncol. 2010;12(6):559 -569.
  16. Al-Saffar NM, Jackson LE, Raynaud FI, et al. The phosphoinositide 3-kinase inhibitor PI-103 downregulates choline kinase alpha leading to phosphocholine and total choline decrease detected by magnetic res- onance spectroscopy. Cancer Res. 2010;70(13):5507 -5517.
  17. Beloueche-Babari M, Jackson LE, Al-Saffar NM, et al. Identification of magnetic resonance detectable metabolic changes associated with in- hibition of phosphoinositide 3-kinase signaling in human breast cancer cells. Mol Cancer Ther. 2006;5(1):187 -196.
  18. Ardenkjaer-Larsen JH, Fridlund B, Gram A, et al. Increase in signal-to-noise ratio of .10,000 times in liquid-state NMR. Proc Natl Acad Sci USA. 2003;100(18):10158 -10163.
  19. Chaumeil MM, Ozawa T, Park I, et al. Hyperpolarized (13)C MR spec- troscopic imaging can be used to monitor Everolimus treatment in vivo in an orthotopic rodent model of glioblastoma. Neuroimage. 2012;59(1):193 -201.
  20. Dafni H, Larson PE, Hu S, et al. Hyperpolarized 13C spectroscopic imaging informs on hypoxia-inducible factor-1 and myc activity down- stream of platelet-derived growth factor receptor. Cancer Res. 2010;70(19):7400 -7410.
  21. Ward CS, Venkatesh HS, Chaumeil MM, et al. Noninvasive Detection of Target Modulation following Phosphatidylinositol 3-Kinase Inhibition Using Hyperpolarized 13C Magnetic Resonance Spectroscopy. Cancer Res. 2010;70(4):1296 -1305.
  22. Pouyssegur J, Dayan F, Mazure NM. Hypoxia signalling in cancer and approaches to enforce tumour regression. Nature. 2006;441(7092):437 -443.
  23. Kroemer G, Pouyssegur J. Tumor cell metabolism: cancer's Achilles' heel. Cancer Cell. 2008;13(6):472 -482.
  24. Glunde K, Shah T, Winnard PT, Jr, et al. Hypoxia regulates choline kinase expression through hypoxia-inducible factor-1 alpha signaling in a human prostate cancer model. Cancer Res. 2008;68(1):172 -180.
  25. Semenza GL, Roth PH, Fang HM, Wang GL. Transcriptional regulation of genes encoding glycolytic enzymes by hypoxia-inducible factor 1. J Biol Chem. 1994;269(38):23757 -23763.
  26. Firth JD, Ebert BL, Ratcliffe PJ. Hypoxic regulation of lactate dehydro- genase A. Interaction between hypoxia-inducible factor 1 and cAMP re- sponse elements. J Biol Chem. 1995;270(36):21021 -21027.
  27. Gunther HS, Schmidt NO, Phillips HS, et al. Glioblastoma-derived stem cell-enriched cultures form distinct subgroups according to molecular and phenotypic criteria. Oncogene. 2008;27(20):2897 -2909.
  28. Sarkaria JN, Yang L, Grogan PT, et al. Identification of molecular char- acteristics correlated with glioblastoma sensitivity to EGFR kinase inhib- ition through use of an intracranial xenograft test panel. Mol Cancer Ther. 2007;6(3):1167 -1174.
  29. Yang L, Clarke MJ, Carlson BL, et al. PTEN loss does not predict for re- sponse to RAD001 (Everolimus) in a glioblastoma orthotopic xenograft test panel. Clin Cancer Res. 2008;14(12):3993 -4001.
  30. Iorio E, Mezzanzanica D, Alberti P, et al. Alterations of choline phospho- lipid metabolism in ovarian tumor progression. Cancer Res. 2005;65(20):9369 -9376.
  31. Gabellieri C, Beloueche-Babari M, Jamin Y, Payne GS, Leach MO, Eykyn TR. Modulation of choline kinase activity in human cancer cells observed by dynamic 31P NMR. NMR Biomed. 2009;22(4):456 -461.
  32. Vassault A. Lactate dehydrogenase. in Methods of Enzymatic Analysis. 1983;3.
  33. Ronen SM, Rushkin E, Degani H. Lipid metabolism in T47D human breast cancer cells: 31P and 13C-NMR studies of choline and ethanolamine uptake. Biochim Biophys Acta. 1991;1095(1): 5 -16.
  34. Albers MJ, Bok R, Chen AP, et al. Hyperpolarized 13C lactate, pyruvate, and alanine: noninvasive biomarkers for prostate cancer detection and grading. Cancer Res. 2008;68(20):8607 -8615.
  35. Kohler SJ, Yen Y, Wolber J, et al. In vivo 13 carbon metabolic imaging at 3T with hyperpolarized 13C-1-pyruvate. Magn Reson Med. 2007;58(1):65 -69.
  36. Brandes AH, Ward CS, Ronen SM. 17-allyamino-17-demethoxygelda- namycin treatment results in a magnetic resonance spectroscopy- detectable elevation in choline-containing metabolites associated with increased expression of choline transporter SLC44A1 and phospholipase A2. Breast Cancer Res. 2010;12(5):R84.
  37. Gingras AC, Raught B, Sonenberg N. Regulation of translation initiation by FRAP/mTOR. Genes Dev. 2001;15(7):807 -826.
  38. Laughner E, Taghavi P, Chiles K, Mahon PC, Semenza GL. HER2 (neu) signaling increases the rate of hypoxia-inducible factor 1alpha (HIF-1alpha) synthesis: novel mechanism for HIF-1-mediated vascular endothelial growth factor expression. Mol Cell Biol. 2001;21(12):3995 -4004.
  39. Harasawa M, Yasuda M, Hirasawa T, et al. Analysis of mTOR inhibition-involved pathway in ovarian clear cell adenocarcinoma. Acta Histochem Cytochem. 2011;44(2):113 -118.
  40. Jiang BH, Jiang G, Zheng JZ, Lu Z, Hunter T, Vogt PK. Phosphatidylinositol 3-kinase signaling controls levels of hypoxia-inducible factor 1. Cell Growth Differ. 2001;12(7):363 -369.
  41. Jordan BF, Black K, Robey IF, Runquist M, Powis G, Gillies RJ. Metabolite changes in HT-29 xenograft tumors following HIF-1alpha inhibition with PX-478 as studied by MR spectroscopy in vivo and ex vivo. NMR Biomed. 2005;18(7):430 -439.
  42. Park I, Bok R, Ozawa T, et al. Detection of early response to temozolomide treatment in brain tumors using hyperpolarized 13C MR metabolic imaging. J Magn Reson Imaging. 2011; 33(6):1284 -1290.
  43. Park I, Jalbert LE, Ozawa T, et al. Hyperpolarized 13C MR Metabolic Imaging Provides an Early Biomarker of MGMT Activity and Response to Temozolomide Treatment. Paper presented at: International Society of Magnetic Resonance in Medicine Annual Meeting; 2011; Montreal, Canada.
  44. Day SE, Kettunen MI, Cherukuri MK, et al. Detecting response of rat C6 glioma tumors to radiotherapy using hyperpolarized [1-13C]pyruvate and 13C magnetic resonance spectroscopic imaging. Magn Reson Med. 2011;65(2):557 -563.
  45. Klomp DW, Wijnen JP, Scheenen TW, Heerschap A. Efficient 1H to 31P polarization transfer on a clinical 3T MR system. Magn Reson Med. 2008;60(6):1298 -1305.
  46. Nelson SJ. Multivoxel magnetic resonance spectroscopy of brain tumors. Mol Cancer Ther. 2003;2(5):497 -507.
  47. Venkatesh et al.: PC and lactate -biomarkers of PI3K inhibition in GBM 324 NEURO-ONCOLOGY † M A R C H 2 0 1 2 at University of California, San Francisco on February 6, 2013 http://neuro-oncology.oxfordjournals.org/ Downloaded from
  48. Bhujwalla ZM, Glickson JD. Detection of tumor response to radiation therapy by in vivo proton MR spectroscopy. Int J Radiat Oncol Biol Phys. 1996;36(3):635 -639.
  49. Park I, Chen AP, Zierhut ML, Ozturk-Isik E, Vigneron DB, Nelson SJ. Implementation of 3 T lactate-edited 3D 1H MR spectroscopic imaging with flyback echo-planar readout for gliomas patients. Ann Biomed Eng. 2011;39(1):193 -204.
  50. Cheng LL, Anthony DC, Comite AR, Black PM, Tzika AA, Gonzalez RG. Quantification of microheterogeneity in glioblastoma multiforme with ex vivo high-resolution magic-angle spinning (HRMAS) proton magnet- ic resonance spectroscopy. Neuro Oncol. 2000;2(2):87 -95.
  51. Park I, Larson PE, Zierhut ML, et al. Hyperpolarized 13C magnetic res- onance metabolic imaging: application to brain tumors. Neuro Oncol. 2010;12(2):133 -144.
  52. Venkatesh et al.: PC and lactate -biomarkers of PI3K inhibition in GBM NEURO-ONCOLOGY † M A R C H 2 0 1 2 325