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Open Access 01-12-2012 | Research

NVP-BEZ235 and NVP-BGT226, dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitors, enhance tumor and endothelial cell radiosensitivity

Authors: Emmanouil Fokas, Michio Yoshimura, Remko Prevo, Geoff Higgins, Wolfgang Hackl, Sauveur-Michel Maira, Eric J Bernhard, WGillies McKenna, Ruth J Muschel

Published in: Radiation Oncology | Issue 1/2012

Abstract

Background

The phosphatidylinositol 3-kinase (PI3K)/Akt pathway is activated in tumor cells and promotes tumor cell survival after radiation-induced DNA damage. Because the pathway may not be completely inhibited after blockade of PI3K itself, due to feedback through mammalian target of rapamycin (mTOR), more effective inhibition might be expected by targeting both PI3K and mTOR inhibition.

Materials and methods

We investigated the effect of two dual PI3K/mTOR (both mTORC1 and mTORC2) inhibitors, NVP-BEZ235 and NVP-BGT226, on SQ20B laryngeal and FaDu hypopharyngeal cancer cells characterised by EGFR overexpression, on T24 bladder tumor cell lines with H-Ras mutation and on endothelial cells. Analysis of target protein phosphorylation, clonogenic survival, number of residual γH2AX foci, cell cycle and apoptosis after radiation was performed in both tumor and endothelial cells. In vitro angiogenesis assays were conducted as well.

Results

Both compounds effectively inhibited phosphorylation of Akt, mTOR and S6 target proteins and reduced clonogenic survival in irradiated tumor cells. Persistence of DNA damage, as evidenced by increased number of γH2AX foci, was detected after irradiation in the presence of PI3K/mTOR inhibition, together with enhanced G2 cell cycle delay. Treatment with one of the inhibitors, NVP-BEZ235, also resulted in decreased clonogenicity after irradiation of tumor cells under hypoxic conditions. In addition, NVP-BEZ235 blocked VEGF- and IR-induced Akt phosphorylation and increased radiation killing in human umbilical venous endothelial cells (HUVEC) and human dermal microvascular dermal cells (HDMVC). NVP-BEZ235 inhibited VEGF-induced cell migration and capillary tube formation in vitro and enhanced the antivascular effect of irradiation. Treatment with NVP-BEZ235 moderately increased apoptosis in SQ20B and HUVEC cells but not in FaDu cells, and increased necrosis in both tumor and endothelial all cells tumor.

Conclusions

The results of this study demonstrate that PI3K/mTOR inhibitors can enhance radiation-induced killing in tumor and endothelial cells and may be of benefit when combined with radiotherapy.

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Begg AC, Stewart FA, Vens C: Strategies to improve radiotherapy with targeted drugs. Nat Rev Cancer 2011, 11: 239-253. 10.1038/nrc3007CrossRefPubMed

Maity A, Bernhard EJ: Modulating tumor vasculature through signaling inhibition to improve cytotoxic therapy. Cancer Res 2010, 70: 2141-2145. 10.1158/0008-5472.CAN-09-3615PubMedCentralCrossRefPubMed

McKenna WG, Muschel RJ, Gupta AK, Hahn SM, Bernhard EJ: The RAS signal transduction pathway and its role in radiation sensitivity. Oncogene 2003, 22: 5866-5875. 10.1038/sj.onc.1206699CrossRefPubMed

Bernhard EJ, Stanbridge EJ, Gupta S, Gupta AK, Soto D, Bakanauskas VJ, Cerniglia GJ, Muschel RJ, McKenna WG: Direct evidence for the contribution of activated N-ras and K-ras oncogenes to increased intrinsic radiation resistance in human tumor cell lines. Cancer Res 2000, 60: 6597-6600.PubMed

Gupta AK, Bakanauskas VJ, Cerniglia GJ, Cheng Y, Bernhard EJ, Muschel RJ, McKenna WG: The Ras radiation resistance pathway. Cancer Res 2001, 61: 4278-4282.PubMed

Kim IA, Fernandes AT, Gupta AK, McKenna WG, Bernhard EJ: The influence of Ras pathway signaling on tumor radiosensitivity. Cancer Metastasis Rev 2004, 23: 227-236.CrossRefPubMed

Prevo R, Deutsch E, Sampson O, Diplexcito J, Cengel K, Harper J, O'Neill P, McKenna WG, Patel S, Bernhard EJ: Class I PI3 kinase inhibition by the pyridinylfuranopyrimidine inhibitor PI-103 enhances tumor radiosensitivity. Cancer Res 2008, 68: 5915-5923. 10.1158/0008-5472.CAN-08-0757CrossRefPubMed

Rudner J, Ruiner CE, Handrick R, Eibl HJ, Belka C, Jendrossek V: The Akt-inhibitor Erufosine induces apoptotic cell death in prostate cancer cells and increases the short term effects of ionizing radiation. Radiat Oncol 2010, 5: 108. 10.1186/1748-717X-5-108PubMedCentralCrossRefPubMed

Cengel KA, Voong KR, Chandrasekaran S, Maggiorella L, Brunner TB, Stanbridge E, Kao GD, McKenna WG, Bernhard EJ: Oncogenic K-Ras signals through epidermal growth factor receptor and wild-type H-Ras to promote radiation survival in pancreatic and colorectal carcinoma cells. Neoplasia 2007, 9: 341-348. 10.1593/neo.06823PubMedCentralCrossRefPubMed

10.

Brunner TB, Cengel KA, Hahn SM, Wu J, Fraker DL, McKenna WG, Bernhard EJ: Pancreatic cancer cell radiation survival and prenyltransferase inhibition: the role of K-Ras. Cancer Res 2005, 65: 8433-8441. 10.1158/0008-5472.CAN-05-0158CrossRefPubMed

11.

Li HF, Kim JS, Waldman T: Radiation-induced Akt activation modulates radioresistance in human glioblastoma cells. Radiat Oncol 2009, 4: 43. 10.1186/1748-717X-4-43PubMedCentralCrossRefPubMed

12.

McKenna WG, Muschel RJ, Gupta AK, Hahn SM, Bernhard EJ: Farnesyltransferase inhibitors as radiation sensitizers. Semin Radiat Oncol 2002, 12: 27-32. 10.1053/srao.2002.34866CrossRefPubMed

13.

Bonner JA, Harari PM, Giralt J, Azarnia N, Shin DM, Cohen RB, Jones CU, Sur R, Raben D, Jassem J, et al.: Radiotherapy plus cetuximab for squamous-cell carcinoma of the head and neck. N Engl J Med 2006, 354: 567-578. 10.1056/NEJMoa053422CrossRefPubMed

14.

Seeliger H, Guba M, Kleespies A, Jauch KW, Bruns CJ: Role of mTOR in solid tumor systems: a therapeutical target against primary tumor growth, metastases, and angiogenesis. Cancer Metastasis Rev 2007, 26: 611-621. 10.1007/s10555-007-9077-8CrossRefPubMed

15.

Manegold PC, Paringer C, Kulka U, Krimmel K, Eichhorn ME, Wilkowski R, Jauch KW, Guba M, Bruns CJ: Antiangiogenic therapy with mammalian target of rapamycin inhibitor RAD001 (Everolimus) increases radiosensitivity in solid cancer. Clin Cancer Res 2008, 14: 892-900. 10.1158/1078-0432.CCR-07-0955CrossRefPubMed

16.

Shinohara ET, Cao C, Niermann K, Mu Y, Zeng F, Hallahan DE, Lu B: Enhanced radiation damage of tumor vasculature by mTOR inhibitors. Oncogene 2005, 24: 5414-5422. 10.1038/sj.onc.1208715CrossRefPubMed

17.

Carracedo A, Ma L, Teruya-Feldstein J, Rojo F, Salmena L, Alimonti A, Egia A, Sasaki AT, Thomas G, Kozma SC, et al.: Inhibition of mTORC1 leads to MAPK pathway activation through a PI3K-dependent feedback loop in human cancer. J Clin Invest 2008, 118: 3065-3074.PubMedCentralPubMed

18.

Maira SM, Stauffer F, Brueggen J, Furet P, Schnell C, Fritsch C, Brachmann S, Chene P, De Pover A, Schoemaker K, et al.: Identification and characterization of NVP-BEZ235, a new orally available dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitor with potent in vivo antitumor activity. Mol Cancer Ther 2008, 7: 1851-1863. 10.1158/1535-7163.MCT-08-0017CrossRefPubMed

19.

Cho DC, Cohen MB, Panka DJ, Collins M, Ghebremichael M, Atkins MB, Signoretti S, Mier JW: The efficacy of the novel dual PI3-kinase/mTOR inhibitor NVP-BEZ235 compared with rapamycin in renal cell carcinoma. Clin Cancer Res 2010, 16: 3628-3638. 10.1158/1078-0432.CCR-09-3022PubMedCentralCrossRefPubMed

20.

Reaper PM, Griffiths MR, Long JM, Charrier JD, Maccormick S, Charlton PA, Golec JM, Pollard JR: Selective killing of ATM- or p53-deficient cancer cells through inhibition of ATR. Nat Chem Biol 2011, 7: 428-430.CrossRefPubMed

21.

Abid MR, Guo S, Minami T, Spokes KC, Ueki K, Skurk C, Walsh K, Aird WC: Vascular endothelial growth factor activates PI3K/Akt/forkhead signaling in endothelial cells. Arterioscler Thromb Vasc Biol 2004, 24: 294-300.CrossRefPubMed

22.

Kim DW, Huamani J, Fu A, Hallahan DE: Molecular strategies targeting the host component of cancer to enhance tumor response to radiation therapy. Int J Radiat Oncol Biol Phys 2006, 64: 38-46. 10.1016/j.ijrobp.2005.02.008CrossRefPubMed

23.

Wood JM, Bold G, Buchdunger E, Cozens R, Ferrari S, Frei J, Hofmann F, Mestan J, Mett H, O'Reilly T, et al.: PTK787/ZK 222584, a novel and potent inhibitor of vascular endothelial growth factor receptor tyrosine kinases, impairs vascular endothelial growth factor-induced responses and tumor growth after oral administration. Cancer Res 2000, 60: 2178-2189.PubMed

24.

Pore N, Gupta AK, Cerniglia GJ, Jiang Z, Bernhard EJ, Evans SM, Koch CJ, Hahn SM, Maity A: Nelfinavir down-regulates hypoxia-inducible factor 1alpha and VEGF expression and increases tumor oxygenation: implications for radiotherapy. Cancer Res 2006, 66: 9252-9259. 10.1158/0008-5472.CAN-06-1239CrossRefPubMed

25.

Wang X, Yue P, Chan CB, Ye K, Ueda T, Watanabe-Fukunaga R, Fukunaga R, Fu H, Khuri FR, Sun SY: Inhibition of mammalian target of rapamycin induces phosphatidylinositol 3-kinase-dependent and Mnk-mediated eukaryotic translation initiation factor 4E phosphorylation. Mol Cell Biol 2007, 27: 7405-7413. 10.1128/MCB.00760-07PubMedCentralCrossRefPubMed

26.

Liu TJ, Koul D, LaFortune T, Tiao N, Shen RJ, Maira SM, Garcia-Echevrria C, Yung WK: NVP-BEZ235, a novel dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitor, elicits multifaceted antitumor activities in human gliomas. Mol Cancer Ther 2009, 8: 2204-2210. 10.1158/1535-7163.MCT-09-0160PubMedCentralCrossRefPubMed

27.

Marone R, Erhart D, Mertz AC, Bohnacker T, Schnell C, Cmiljanovic V, Stauffer F, Garcia-Echeverria C, Giese B, Maira SM, Wymann MP: Targeting melanoma with dual phosphoinositide 3-kinase/mammalian target of rapamycin inhibitors. Mol Cancer Res 2009, 7: 601-613. 10.1158/1541-7786.MCR-08-0366CrossRefPubMed

28.

Manara MC, Nicoletti G, Zambelli D, Ventura S, Guerzoni C, Landuzzi L, Lollini PL, Maira SM, Garcia-Echeverria C, Mercuri M, et al.: NVP-BEZ235 as a new therapeutic option for sarcomas. Clin Cancer Res 2010, 16: 530-540. 10.1158/1078-0432.CCR-09-0816CrossRefPubMed

29.

Konstantinidou G, Bey EA, Rabellino A, Schuster K, Maira MS, Gazdar AF, Amici A, Boothman DA, Scaglioni PP: Dual phosphoinositide 3-kinase/mammalian target of rapamycin blockade is an effective radiosensitizing strategy for the treatment of non-small cell lung cancer harboring K-RAS mutations. Cancer Res 2009, 69: 7644-7652. 10.1158/0008-5472.CAN-09-0823PubMedCentralCrossRefPubMed

30.

Chiarini F, Grimaldi C, Ricci F, Tazzari PL, Evangelisti C, Ognibene A, Battistelli M, Falcieri E, Melchionda F, Pession A, et al.: Activity of the novel dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitor NVP-BEZ235 against T-cell acute lymphoblastic leukemia. Cancer Res 2010, 70: 8097-8107. 10.1158/0008-5472.CAN-10-1814CrossRefPubMed

31.

Vispe S, Cazaux C, Lesca C, Defais M: Overexpression of Rad51 protein stimulates homologous recombination and increases resistance of mammalian cells to ionizing radiation. Nucleic Acids Res 1998, 26: 2859-2864. 10.1093/nar/26.12.2859PubMedCentralCrossRefPubMed

32.

Phung TL, Ziv K, Dabydeen D, Eyiah-Mensah G, Riveros M, Perruzzi C, Sun J, Monahan-Earley RA, Shiojima I, Nagy JA, et al.: Pathological angiogenesis is induced by sustained Akt signaling and inhibited by rapamycin. Cancer Cell 2006, 10: 159-170. 10.1016/j.ccr.2006.07.003PubMedCentralCrossRefPubMed

33.

Dayanir V, Meyer RD, Lashkari K, Rahimi N: Identification of tyrosine residues in vascular endothelial growth factor receptor-2/FLK-1 involved in activation of phosphatidylinositol 3-kinase and cell proliferation. J Biol Chem 2001, 276: 17686-17692. 10.1074/jbc.M009128200CrossRefPubMed

34.

Sun JF, Phung T, Shiojima I, Felske T, Upalakalin JN, Feng D, Kornaga T, Dor T, Dvorak AM, Walsh K, Benjamin LE: Microvascular patterning is controlled by fine-tuning the Akt signal. Proc Natl Acad Sci USA 2005, 102: 128-133. 10.1073/pnas.0403198102PubMedCentralCrossRefPubMed

35.

Raynaud FI, Eccles S, Clarke PA, Hayes A, Nutley B, Alix S, Henley A, Di-Stefano F, Ahmad Z, Guillard S, et al.: Pharmacologic characterization of a potent inhibitor of class I phosphatidylinositide 3-kinases. Cancer Res 2007, 67: 5840-5850. 10.1158/0008-5472.CAN-06-4615CrossRefPubMed

36.

Fokas E, Im JH, Hill S, Yameen S, Stratford M, Beech J, Hackl W, Maira SM, Bernhard EJ, McKenna WG, Muschel RJ: Dual inhibition of the PI3K/mTOR pathway increases tumor radiosensitivity by normalizing tumor vasculature. Cancer Res 2012, 72: 239-248. 10.1158/0008-5472.CAN-11-2263CrossRefPubMed

Title: NVP-BEZ235 and NVP-BGT226, dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitors, enhance tumor and endothelial cell radiosensitivity
Authors: Emmanouil Fokas
Michio Yoshimura
Remko Prevo
Geoff Higgins
Wolfgang Hackl
Sauveur-Michel Maira
Eric J Bernhard
WGillies McKenna
Ruth J Muschel
Publication date: 01-12-2012
Publisher: BioMed Central
Published in: Radiation Oncology / Issue 1/2012
Electronic ISSN: 1748-717X
DOI: https://doi.org/10.1186/1748-717X-7-48

Keynote webinar | Spotlight on medication adherence

Springer Medicine

NVP-BEZ235 and NVP-BGT226, dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitors, enhance tumor and endothelial cell radiosensitivity

Abstract

Background

Materials and methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Materials and methods

Results

Conclusions

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