Cancer Chemotherapy and Pharmacology

, Volume 71, Issue 5, pp 1297–1307 | Cite as

Benchmarking effects of mTOR, PI3K, and dual PI3K/mTOR inhibitors in hepatocellular and renal cell carcinoma models developing resistance to sunitinib and sorafenib

  • Maria Serova
  • Armand de Gramont
  • Annemilaï Tijeras-Raballand
  • Célia Dos Santos
  • Maria Eugenia Riveiro
  • Khemaies Slimane
  • Sandrine Faivre
  • Eric Raymond
Original Article

Abstract

Purpose

To evaluate first-generation rapamycin analogs (everolimus, temsirolimus, and rapamycin) and second-generation drugs inhibiting mTOR kinase (AZD-8055), PI3K (BKM-120) or both (BEZ-235 and GDC-0980) in hepatocellular carcinoma (HCC) and renal cell carcinoma (RCC) cells characterized for acquired resistance to sorafenib or sunitinib.

Methods

Anti-proliferative (MTT assay) and cell signaling (Western blot) effects of rapamycin analogs (1–20 μM) and second-generation drugs (0.03–20.0 μM) were assessed in human HCC SK-HEP1, RCC 786-0, and sorafenib- (SK-Sora) or sunitinib-resistant (786-Suni) cells.

Results

In SK-HEP1 cells displaying high PTEN and Bcl2 expression, rapamycin analogs had poor anti-proliferative effects. However, SK-Sora cells were more sensitive to rapamycin analogs (≥1 μM) than SK-HEP1 cells. In 786-0 cells, lacking PTEN and Bcl2 expression, ≥1 μM rapamycin analogs blocked mTORC1 signaling, transiently activated Akt, and inhibited cell proliferation. Protracted sunitinib exposure in 786-Suni cells yielded an increase in p27 expression and a decreased sensitivity to rapamycin analogs, although mTORC1 function could be inhibited with rapamycin analogs. Second-generation drugs induced more potent growth inhibition than rapamycin analogs at concentrations >0.03 μM in parental cells, SK-Sora, and 786-Suni cells. Growth inhibitory concentrations of these new drugs also blocked mTORC1 downstream targets.

Conclusions

Rapamycin analogs inhibited mTORC1 downstream targets and yielded anti-proliferative effects in HCC and RCC cells. Second-generation drugs also appeared to be potent inhibitors of mTORC1 signaling; however, they appeared to be far more potent in inhibiting cellular proliferation in parental HCC and RCC cells and in cells developing resistance to sorafenib or sunitinib.

Keywords

Kidney cancer mTOR inhibitor Resistance VEGFr tyrosine kinase inhibitor Angiogenesis 

Supplementary material

280_2013_2129_MOESM1_ESM.docx (13 kb)
Supplementary material 1 (DOCX 12 kb)

References

  1. 1.
    International Agency for Research on Cancer. Globocan 2008 Fast Stats: hepatocellular carcinoma: Worldwide Incidence and Mortality. International Agency for Research on Cancer Web Site. Available at http://globocan.iarc.fr/
  2. 2.
    International Agency for Research on Cancer. Globocan 2008 Fast Stats: Renal Cell Carcinoma: Worldwide Incidence and Mortality. International Agency for Research on Cancer Web Site. Available at http://globocan.iarc.fr/bar_pop.asp?selection=221900&title=World&sex=0&statistic=0&window=1&grid=1&info=1&color1=4&color1e=&color2=5&color2e=&orientation=1&submit=%A0Execute%A0
  3. 3.
    Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, de Oliveira AC, Santoro A, Raoul JL, Forner A, Schwartz M, Porta C, Zeuzem S, Bolondi L, Greten TF, Galle PR, Seitz JF, Borbath I, Haussinger D, Giannaris T, Shan M, Moscovici M, Voliotis D, Bruix J, for the SHARP Investigators Study Group (2008) Sorafenib in advanced hepatocellular carcinoma. N Engl J Med 359:378–390CrossRefGoogle Scholar
  4. 4.
    Motzer RJ, Hutson TE, Tomczak P, Michaelson MD, Bukowski RM, Rixe O, Oudard S, Negrier S, Szczylik C, Kim ST, Chen I, Bycott PW, Baum CM, Figlin RA (2007) Sunitinib versus interferon alfa in metastatic renal-cell carcinoma. N Engl J Med 356:115–124PubMedCrossRefGoogle Scholar
  5. 5.
    Escudier B, Szczylik C, Hutson TE, Demkow T, Staehler M, Rolland F, Negrier S, Laferriere N, Scheuring UJ, Cella D, Shah S, Bukowski RM (2009) Randomized phase II trial of first-line treatment with sorafenib versus interferon alfa-2a in patients with metastatic renal cell carcinoma. J Clin Oncol 27:1280–1289PubMedCrossRefGoogle Scholar
  6. 6.
    Escudier B, Eisen T, Stadler WM, Szczylik C, Oudard S, Staehler M, Negrier S, Chevreau C, Desai AA, Rolland F, Demkow T, Hutson TE, Gore M, Anderson S, Hofilena G, Shan M, Pena C, Lathia C, Bukowski RM (2009) Sorafenib for treatment of renal cell carcinoma: final efficacy and safety results of the phase III treatment approaches in renal cancer global evaluation trial. J Clin Oncol 27:3312–3318PubMedCrossRefGoogle Scholar
  7. 7.
    Meric-Bernstam F, Gonzalez-Angulo AM (2009) Targeting the mTOR signaling network for cancer therapy. J Clin Oncol 27:2278–2287PubMedCrossRefGoogle Scholar
  8. 8.
    Faivre S, Kroemer G, Raymond E (2006) Current development of mTOR inhibitors as anticancer agents. Nat Rev Drug Discov 5:671–688PubMedCrossRefGoogle Scholar
  9. 9.
    Faivre S, Raymond E (2008) Mechanism of action of rapalogues: the antiangiogenic hypothesis. Exp Opin Invest Drugs 17:1619–1621CrossRefGoogle Scholar
  10. 10.
    Keunen O, Johansson M, Oudin A, Sanzey M, Rahim SA, Fack F, Thorsen F, Taxt T, Bartos M, Jirik R, Miletic H, Wang J, Stieber D, Stuhr L, Moen I, Rygh CB, Bjerkvig R, Niclou SP (2011) Anti-VEGF treatment reduces blood supply and increases tumor cell invasion in glioblastoma. Proc Natl Acad Sci USA 108:3749–3754PubMedCrossRefGoogle Scholar
  11. 11.
    Hudes G, Carducci M, Tomczak P, Dutcher J, Figlin R, Kapoor A, Staroslawska E, Sosman J, McDermott D, Bodrogi I, Kovacevic Z, Lesovoy V, Schmidt-Wolf IG, Barbarash O, Gokmen E, O’Toole T, Lustgarten S, Moore L, Motzer RJ (2007) Temsirolimus, interferon alfa, or both for advanced renal-cell carcinoma. N Engl J Med 356:2271–2281PubMedCrossRefGoogle Scholar
  12. 12.
    Escudier B, Kataja V (2010) Renal cell carcinoma: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol 21(suppl 5):v137–v139PubMedCrossRefGoogle Scholar
  13. 13.
    Ljungberg B, Cowan NC, Hanbury DC, Hora M, Kuczyk MA, Merseburger AS, Patard JJ, Mulders PF, Sinescu IC (2010) EAU guidelines on renal cell carcinoma: the 2010 update. Eur Urol 58:398–406PubMedCrossRefGoogle Scholar
  14. 14.
    de Reijke TM, Bellmunt J, van Poppel H, Marreaud S, Aapro M (2009) EORTC-GU group expert opinion on metastatic renal cell cancer. Eur J Cancer 45:765–773PubMedCrossRefGoogle Scholar
  15. 15.
    National Comprehensive Cancer Network. NCCN Clinical Practice Guidelines in Oncology™ Kidney Cancer (Version 2.2012). 2012. Available at http://www.nccn.org/professionals/physician_gls/pdf/kidney.pdf
  16. 16.
    Motzer RJ, Escudier B, Oudard S, Hutson TE, Porta C, Bracarda S, Grunwald V, Thompson JA, Figlin RA, Hollaender N, Kay A, Ravaud A (2010) Phase 3 trial of everolimus for metastatic renal cell carcinoma: final results and analysis of prognostic factors. Cancer 116:4256–4265PubMedCrossRefGoogle Scholar
  17. 17.
    U.S. National Institutes of Health. ClinicalTrials.Gov Web Site. Available at http://www.clinicaltrials.gov/ct2/home. Accessed 23 July 2012
  18. 18.
    Fasolo A, Sessa C (2012) Targeting mTOR pathways in human malignancies. Curr Pharm Des 18:2766–2777PubMedCrossRefGoogle Scholar
  19. 19.
    Delbaldo C, Albert S, Dreyer C, Sablin MP, Serova M, Raymond E, Faivre S (2011) Predictive biomarkers for the activity of mammalian target of rapamycin (mTOR) inhibitors. Target Oncol 6:119–124PubMedCrossRefGoogle Scholar
  20. 20.
    Neshat MS, Mellinghoff IK, Tran C, Stiles B, Thomas G, Petersen R, Frost P, Gibbons JJ, Wu H, Sawyers CL (2001) Enhanced sensitivity of PTEN-deficient tumors to inhibition of FRAP/mTOR. Proc Natl Acad Sci USA 98:10314–10319PubMedCrossRefGoogle Scholar
  21. 21.
    Veilleux A, Houde VP, Bellmann K, Marette A (2010) Chronic inhibition of the mTORC1/S6K1 pathway increases insulin-induced PI3K activity but inhibits Akt2 and glucose transport stimulation in 3T3-L1 adipocytes. Mol Endocrinol 24:766–778PubMedCrossRefGoogle Scholar
  22. 22.
    Aguirre D, Boya P, Bellet D, Faivre S, Troalen F, Benard J, Saulnier P, Hopkins-Donaldson S, Zangemeister-Wittke U, Kroemer G, Raymond E (2004) Bcl-2 and CCND1/CDK4 expression levels predict the cellular effects of mTOR inhibitors in human ovarian carcinoma. Apoptosis 9:797–805PubMedCrossRefGoogle Scholar
  23. 23.
    Hidalgo M, Buckner JC, Erlichman C, Pollack MS, Boni JP, Dukart G, Marshall B, Speicher L, Moore L, Rowinsky EK (2006) A phase I and pharmacokinetic study of temsirolimus (CCI-779) administered intravenously daily for 5 days every 2 weeks to patients with advanced cancer. Clin Cancer Res 12:5755–5763PubMedCrossRefGoogle Scholar
  24. 24.
    O’Donnell A, Faivre S, Burris HA III, Rea D, Papadimitrakopoulou V, Shand N, Lane HA, Hazell K, Zoellner U, Kovarik JM, Brock C, Jones S, Raymond E, Judson I (2008) Phase I pharmacokinetic and pharmacodynamic study of the oral mammalian target of rapamycin inhibitor everolimus in patients with advanced solid tumors. J Clin Oncol 26:1588–1595PubMedCrossRefGoogle Scholar
  25. 25.
    Vilar E, Perez-Garcia J, Tabernero J (2011) Pushing the envelope in the mTOR pathway: the second generation of inhibitors. Mol Cancer Ther 10:395–403PubMedCrossRefGoogle Scholar
  26. 26.
    Chresta CM, Davies BR, Hickson I, Harding T, Cosulich S, Critchlow SE, Vincent JP, Ellston R, Jones D, Sini P, James D, Howard Z, Dudley P, Hughes G, Smith L, Maguire S, Hummersone M, Malagu K, Menear K, Jenkins R, Jacobsen M, Smith GC, Guichard S, Pass M (2010) AZD8055 is a potent, selective, and orally bioavailable ATP-competitive mammalian target of rapamycin kinase inhibitor with in vitro and in vivo antitumor activity. Cancer Res 70:288–298PubMedCrossRefGoogle Scholar
  27. 27.
    Cho DC, Cohen MB, Panka DJ, Collins M, Ghebremichael M, Atkins MB, Signoretti S, Mier JW (2010) The efficacy of the novel dual PI3-kinase/mTOR inhibitor NVP-BEZ235 compared with rapamycin in renal cell carcinoma. Clin Cancer Res 16:3628–3638PubMedCrossRefGoogle Scholar
  28. 28.
    Dolly S, Wagner AJ, Bendell JC, Yan Y, Ware JA, Mazina KE, Holden SN, Derynck MK, De Bono JS, Burris HA III (2010) A first-in-human, phase l study to evaluate the dual PI3K/mTOR inhibitor GDC-0980 administered QD in patients with advanced solid tumors or non-Hodgkin’s lymphoma [abstract]. J Clin Oncol 28(May 20 suppl):3079Google Scholar
  29. 29.
    Maira SM, Pecchi S, Huang A, Burger M, Knapp M, Sterker D, Schnell C, Guthy D, Nagel T, Wiesmann M, Brachmann S, Fritsch C, Dorsch M, Chene P, Shoemaker K, De Pover A, Menezes D, Martiny-Baron G, Fabbro D, Wilson CJ, Schlegel R, Hofmann F, Garcia-Echeverria C, Sellers WR, Voliva CF (2012) Identification and characterization of NVP-BKM120, an orally available pan-class I PI3-kinase inhibitor. Mol Cancer Ther 11:317–328PubMedCrossRefGoogle Scholar
  30. 30.
    Piguet AC, Saar B, Hlushchuk R, St-Pierre MV, McSheehy PM, Radojevic V, Afthinos M, Terracciano L, Djonov V, Dufour JF (2011) Everolimus augments the effects of sorafenib in a syngeneic orthotopic model of hepatocellular carcinoma. Mol Cancer Ther 10:1007–1017PubMedCrossRefGoogle Scholar
  31. 31.
    Patel PH, Senico PL, Curiel RE, Motzer RJ (2009) Phase I study combining treatment with temsirolimus and sunitinib malate in patients with advanced renal cell carcinoma. Clin Genitourin Cancer 7:24–27PubMedCrossRefGoogle Scholar
  32. 32.
    Molina AM, Feldman DR, Voss MH, Ginsberg MS, Baum MS, Brocks DR, Fischer PM, Trinos MJ, Patil S, Motzer RJ (2012) Phase 1 trial of everolimus plus sunitinib in patients with metastatic renal cell carcinoma. Cancer 118:1868–1876PubMedCrossRefGoogle Scholar
  33. 33.
    Roulin D, Waselle L, Dormond-Meuwly A, Dufour M, Demartines N, Dormond O (2011) Targeting renal cell carcinoma with NVP-BEZ235, a dual PI3K/mTOR inhibitor, in combination with sorafenib. Mol Cancer 10:90PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Maria Serova
    • 1
    • 2
  • Armand de Gramont
    • 1
  • Annemilaï Tijeras-Raballand
    • 1
    • 2
  • Célia Dos Santos
    • 1
    • 2
  • Maria Eugenia Riveiro
    • 2
  • Khemaies Slimane
    • 3
  • Sandrine Faivre
    • 2
  • Eric Raymond
    • 2
  1. 1.AAREC Filia ResearchBoulogne-BillancourtFrance
  2. 2.INSERM U728 and Department of Medical OncologyBeaujon University Hospital (AP-HP–PRES Paris 7 Diderot)ClichyFrance
  3. 3.Novartis Pharma S.A.S.Rueil-MalmaisonFrance

Personalised recommendations