Journal of Neuro-Oncology

, Volume 120, Issue 3, pp 473–481 | Cite as

Profiling Hsp90 differential expression and the molecular effects of the Hsp90 inhibitor IPI-504 in high-grade glioma models

  • Kaijun Di
  • Stephen T. Keir
  • Daniela Alexandru-Abrams
  • Xing Gong
  • Howard Nguyen
  • Henry S. Friedman
  • Daniela A. Bota
Laboratory Investigation


Retaspimycin hydrochloride (IPI-504), an Hsp90 (heat shock protein 90) inhibitor, has shown activity in multiple preclinical cancer models, such as lung, breast and ovarian cancers. However, its biological effects in gliomas and normal brain derived cellular populations remain unknown. In this study, we profiled the expression pattern of Hsp90α/β mRNA in stable glioma cell lines, multiple glioma-derived primary cultures and human neural stem/progenitor cells. The effects of IPI-504 on cell proliferation, apoptosis, motility and expression of Hsp90 client proteins were evaluated in glioma cell lines. In vivo activity of IPI-504 was investigated in subcutaneous glioma xenografts. Our results showed Hsp90α and Hsp90β expression levels to be patient-specific, higher in high-grade glioma-derived primary cells than in low-grade glioma-derived primary cells, and strongly correlated with CD133 expression and differentiation status of cells. Hsp90 inhibition by IPI-504 induced apoptosis, blocked migration and invasion, and significantly decreased epidermal growth factor receptor levels, mitogen-activated protein kinase and/or Akt activities, and secretion of vascular endothelial growth factor in glioma cell lines. In vivo study showed that IPI-504 could mildly attenuate tumor growth in immunocompromised mice. These findings suggest that targeting Hsp90 by IPI-504 has the potential to become an active therapeutic strategy in gliomas in a selective group of patients, but further research into combination therapies is still needed.


Hsp90 IPI-504 Glioma Stem cells Anti-tumor activity 



We thank Infinity Pharmaceuticals for providing IPI-504 and Dr. Julian Adams for his suggestions and comments. We gratefully acknowledge Dr. David A. Fruman for providing us phospho-AKT (Ser473) antibody, and the Core Facility of Department of Pathology & Laboratory Medicine of UC Irvine for help performing the immunocytochemistry experiments. This study was supported in part by research funds donated by Ralph and Suzanne Stern, start-up funds to Dr. Bota from the UC Irvine, and the National Cancer Institute of the National Institutes of Health under Award Number P30CA062203.

Conflict of interest

No potential conflicts of interest were disclosed.

Supplementary material

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Supplementary material 1 (DOC 67 kb)
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Supplementary material 6 (DOC 36 kb)


  1. 1.
    Morimoto RI, Kline MP, Bimston DN, Cotto JJ (1997) The heat-shock response: regulation and function of heat-shock proteins and molecular chaperones. Essays Biochem 32:17–29PubMedGoogle Scholar
  2. 2.
    Lindquist S, Craig EA (1988) The Heat-Shock Proteins. Annu Rev Genet 22:631–677PubMedCrossRefGoogle Scholar
  3. 3.
    Millson SH, Truman AW, Racz A, Hu B, Panaretou B, Nuttall J et al (2007) Expressed as the sole Hsp90 of yeast, the alpha and beta isoforms of human Hsp90 differ with regard to their capacities for activation of certain client proteins, whereas only Hsp90 beta generates sensitivity to the Hsp90 inhibitor radicicol. FEBS J 274:4453–4463PubMedCrossRefGoogle Scholar
  4. 4.
    Jackson SE (2013) Hsp90: structure and function. Top Curr Chem 328:155–240PubMedCrossRefGoogle Scholar
  5. 5.
    Cheng Q, Chang JT, Geradts J, Neckers LM, Haystead T, Spector NL et al (2012) Amplification and high-level expression of heat shock protein 90 marks aggressive phenotypes of human epidermal growth factor receptor 2 negative breast cancer. Breast Cancer Res 14:R62PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    Wang J, Cui S, Zhang X, Wu Y, Tang H (2013) High expression of heat shock protein 90 is associated with tumor aggressiveness and poor prognosis in patients with advanced gastric cancer. PLoS One 8:e62876PubMedCentralPubMedCrossRefGoogle Scholar
  7. 7.
    Khalil AA, Kabapy NF, Deraz SF, Smith C (2011) Heat shock proteins in oncology: diagnostic biomarkers or therapeutic targets? Biochim Biophys Acta 1816:89–104PubMedGoogle Scholar
  8. 8.
    Den RB, Lu B (2012) Heat shock protein 90 inhibition: rationale and clinical potential. Ther Adv Med Oncol 4:211–218PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Roe SM, Prodromou C, O’Brien R, Ladbury JE, Piper PW, Pearl LH (1999) Structural basis for inhibition of the Hsp90 molecular chaperone by the antitumor antibiotics radicicol and geldanamycin. J Med Chem 42:260–266PubMedCrossRefGoogle Scholar
  10. 10.
    Sydor JR, Normant E, Pien CS, Porter JR, Ge J, Grenier L et al (2006) Development of 17-allylamino-17-demethoxygeldanamycin hydroquinone hydrochloride (IPI-504), an anti-cancer agent directed against Hsp90. Proc Natl Acad Sci USA 103:17408–17413PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Ge J, Normant E, Porter JR, Ali JA, Dembski MS, Gao Y et al (2006) Design, synthesis, and biological evaluation of hydroquinone derivatives of 17-amino-17-demethoxygeldanamycin as potent, water-soluble inhibitors of Hsp90. J Med Chem 49:4606–4615PubMedCrossRefGoogle Scholar
  12. 12.
    Maroney AC, Marugan JJ, Mezzasalma TM, Barnakov AN, Garrabrant TA, Weaner LE et al (2006) Dihydroquinone ansamycins: toward resolving the conflict between low in vitro affinity and high cellular potency of geldanamycin derivatives. Biochemistry 45:5678–5685PubMedCrossRefGoogle Scholar
  13. 13.
    Gladson CL, Prayson RA, Liu WM (2010) The pathobiology of glioma tumors. Annu Rev Pathol 5:33–50PubMedCentralPubMedCrossRefGoogle Scholar
  14. 14.
    Wen PY, Kesari S (2008) Malignant gliomas in adults. N Engl J Med 359:492–507PubMedCrossRefGoogle Scholar
  15. 15.
    Siegelin MD, Habel A, Gaiser T (2009) 17-AAG sensitized malignant glioma cells to death-receptor mediated apoptosis. Neurobiol Dis 33:243–249PubMedCrossRefGoogle Scholar
  16. 16.
    Rao RD, Uhm JH, Krishnan S, James CD (2003) Genetic and signaling pathway alterations in glioblastoma: relevance to novel targeted therapies. Front Biosci 8:e270–e280PubMedCrossRefGoogle Scholar
  17. 17.
    Sauvageot CM, Weatherbee JL, Kesari S, Winters SE, Barnes J, Dellagatta J et al (2009) Efficacy of the HSP90 inhibitor 17-AAG in human glioma cell lines and tumorigenic glioma stem cells. Neuro Oncol 11:109–121PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    Singh SK, Clarke ID, Terasaki M, Bonn VE, Hawkins C, Squire J et al (2003) Identification of a cancer stem cell in human brain tumors. Cancer Res 63:5821–5828PubMedGoogle Scholar
  19. 19.
    Singh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T et al (2004) Identification of human brain tumour initiating cells. Nature 432:396–401PubMedCrossRefGoogle Scholar
  20. 20.
    Bao S, Wu Q, McLendon RE, Hao Y, Shi Q, Hjelmeland AB et al (2006) Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature 444:756–760PubMedCrossRefGoogle Scholar
  21. 21.
    Di K, Linskey ME, Bota DA (2013) TRIM11 is overexpressed in high-grade gliomas and promotes proliferation, invasion, migration and glial tumor growth. Oncogene 32:5038–5047PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    Schwartz PH, Bryant PJ, Fuja TJ, Su H, O’Dowd DK, Klassen H (2003) Isolation and characterization of neural progenitor cells from post-mortem human cortex. J Neurosci Res 74:838–851PubMedCrossRefGoogle Scholar
  23. 23.
    Pistollato F, Chen HL, Rood BR, Zhang HZ, D’Avella D, Denaro L et al (2009) Hypoxia and HIF1alpha repress the differentiative effects of BMPs in high-grade glioma. Stem Cells 27:7–17PubMedCrossRefGoogle Scholar
  24. 24.
    Gong X, Schwartz PH, Linskey ME, Bota DA (2011) Neural stem/progenitors and glioma stem-like cells have differential sensitivity to chemotherapy. Neurology 76:1126–1134PubMedCentralPubMedCrossRefGoogle Scholar
  25. 25.
    Bota DA, Alexandru D, Keir ST, Bigner D, Vredenburgh J, Friedman HS (2013) Proteasome inhibition with bortezomib induces cell death in GBM stem-like cells and temozolomide-resistant glioma cell lines, but stimulates GBM stem-like cells’ VEGF production and angiogenesis. J Neurosurg 119:1415–1423PubMedCrossRefGoogle Scholar
  26. 26.
    Keir ST, Dewhirst MW, Kirkpatrick JP, Bigner DD, Batinic-Haberle I (2011) Cellular redox modulator, ortho Mn(III) meso-tetrakis(N-n-hexylpyridinium-2-yl)porphyrin, MnTnHex-2-PyP(5+) in the treatment of brain tumors. Anticancer Agents Med Chem 11:202–212PubMedCentralPubMedCrossRefGoogle Scholar
  27. 27.
    Yamada T, Hashiguchi A, Fukushima S, Kakita Y, Umezawa A, Maruyama T et al (2000) Function of 90-kDa heat shock protein in cellular differentiation of human embryonal carcinoma cells. In Vitro Cell Dev Biol Anim 36:139–146PubMedCrossRefGoogle Scholar
  28. 28.
    Bradley E, Bieberich E, Mivechi NF, Tangpisuthipongsa D, Wang G (2012) Regulation of embryonic stem cell pluripotency by heat shock protein 90. Stem Cells 30:1624–1633PubMedCentralPubMedCrossRefGoogle Scholar
  29. 29.
    Tarasenko YI, Yu Y, Jordan PM, Bottenstein J, Wu P (2004) Effect of growth factors on proliferation and phenotypic differentiation of human fetal neural stem cells. J Neurosci Res 78:625–636PubMedCrossRefGoogle Scholar
  30. 30.
    Lim DA, Cha S, Mayo MC, Chen MH, Keles E, VandenBerg S et al (2007) Relationship of glioblastoma multiforme to neural stem cell regions predicts invasive and multifocal tumor phenotype. Neuro Oncol 9:424–429PubMedCentralPubMedCrossRefGoogle Scholar
  31. 31.
    Takano S, Yoshii Y, Kondo S, Suzuki H, Maruno T, Shirai S et al (1996) Concentration of vascular endothelial growth factor in the serum and tumor tissue of brain tumor patients. Cancer Res 56:2185–2190PubMedGoogle Scholar
  32. 32.
    Maity A, Pore N, Lee J, Solomon D, O’Rourke DM (2000) Epidermal growth factor receptor transcriptionally up-regulates vascular endothelial growth factor expression in human glioblastoma cells via a pathway involving phosphatidylinositol 3′-kinase and distinct from that induced by hypoxia. Cancer Res 60:5879–5886PubMedGoogle Scholar
  33. 33.
    Song D, Chaerkady R, Tan AC, Garcia-Garcia E, Nalli A, Suarez-Gauthier A et al (2008) Antitumor activity and molecular effects of the novel heat shock protein 90 inhibitor, IPI-504, in pancreatic cancer. Mol Cancer Ther 7:3275–3284PubMedCrossRefGoogle Scholar
  34. 34.
    Fu J, Koul D, Yao J, Wang S, Yuan Y, Colman H et al (2013) Novel HSP90 inhibitor NVP-HSP990 targets cell-cycle regulators to ablate Olig2-positive glioma tumor-initiating cells. Cancer Res 73:3062–3074PubMedCentralPubMedCrossRefGoogle Scholar
  35. 35.
    Newcomb EW, Lukyanov Y, Schnee T, Esencay M, Fischer I, Hong D et al (2007) The geldanamycin analogue 17-allylamino-17-demethoxygeldanamycin inhibits the growth of GL261 glioma cells in vitro and in vivo. Anticancer Drugs 18:875–882PubMedGoogle Scholar
  36. 36.
    Roue G, Perez-Galan P, Mozos A, Lopez-Guerra M, Xargay-Torrent S, Rosich L et al (2011) The Hsp90 inhibitor IPI-504 overcomes bortezomib resistance in mantle cell lymphoma in vitro and in vivo by down-regulation of the prosurvival ER chaperone BiP/Grp78. Blood 117:1270–1279PubMedCrossRefGoogle Scholar
  37. 37.
    De Raedt T, Walton Z, Yecies JL, Li D, Chen Y, Malone CF et al (2011) Exploiting cancer cell vulnerabilities to develop a combination therapy for ras-driven tumors. Cancer Cell 20:400–413PubMedCentralPubMedCrossRefGoogle Scholar
  38. 38.
    Scaltriti M, Serra V, Normant E, Guzman M, Rodriguez O, Lim AR et al (2011) Antitumor activity of the Hsp90 inhibitor IPI-504 in HER2-positive trastuzumab-resistant breast cancer. Mol Cancer Ther 10:817–824PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Kaijun Di
    • 1
  • Stephen T. Keir
    • 2
  • Daniela Alexandru-Abrams
    • 1
  • Xing Gong
    • 3
  • Howard Nguyen
    • 3
  • Henry S. Friedman
    • 2
  • Daniela A. Bota
    • 1
    • 3
    • 4
  1. 1.Department of Neurological SurgeryUC Irvine School of MedicineIrvineUSA
  2. 2.Duke University Medical CenterDurhamUSA
  3. 3.Department of NeurologyUC Irvine School of MedicineIrvineUSA
  4. 4.UC Irvine Chao Family Comprehensive Cancer CenterOrangeUSA

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