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Multiple receptor tyrosine kinases regulate HIF-1α and HIF-2α in normoxia and hypoxia in neuroblastoma: implications for antiangiogenic mechanisms of multikinase inhibitors

Oncogene volume 29, pages 2938–2949 (2010)Cite this article

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Abstract

Novel treatment approaches are needed for children with advanced neuroblastoma. Studies with neuroblastoma cells have indicated the presence of a hypoxia-driven vascular endothelial growth factor (VEGF)/VEGF receptor (VEGFR)-1 autocrine loop modulating hypoxia-inducible factor-1alpha (HIF-1α). Whether other receptor tyrosine kinases (RTKs) are capable of modulating HIF-1α levels and whether RTKs can regulate HIF-2α as well is largely unknown. We evaluated neuroblastoma cell lines for expression of various RTKs. Although cell lines were heterogeneous in the expression of VEGFR-1, -3, c-Kit and RET, most cells expressed PDGFR-α and -β. Ligand-induced activation of multiple RTKs upregulated HIF-1α levels, whereas activation of VEGFR-1 alone upregulated HIF-2α. Multitargeted tyrosine kinase inhibitor sunitinib reduced hypoxia-induced rises in HIF-1α and HIF-2α through mechanisms involving effects on both mRNA levels and protein stability. In addition, sunitinib and sorafenib had direct effects on tumor cell viability in vitro. In a neuroblastoma xenograft model, tumor growth inhibition by sunitinib was associated with inhibition of angiogenesis and reduced HIF-1α levels. These findings show that multiple RTKs may regulate the HIF axis in normoxia and hypoxia and suggest that multikinase inhibitors may exert antiangiogenic effects not only by direct effects on endothelial cells, but also by blocking compensatory hypoxia- and ligand-induced changes in HIF-1α and HIF-2α.

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References

  • Beaudry P, Nilsson M, Rioth M, Prox D, Poon D, Xu L et al. (2008). Potent antitumor effects of ZD6474 on neuroblastoma via dual targeting of tumor cells and tumor endothelium. Mol Cancer Ther 7: 418–424.

    Article  CAS  PubMed  Google Scholar 

  • Beppu K, Jaboine J, Merchant MS, Mackall CL, Thiele CJ . (2004). Effect of imatinib mesylate on neuroblastoma tumorigenesis and vascular endothelial growth factor expression. J Natl Cancer Inst 96: 46–55.

    Article  CAS  PubMed  Google Scholar 

  • Bruns CJ, Solorzano CC, Harbison MT, Ozawa S, Tsan R, Fan D et al. (2000). Blockade of the epidermal growth factor receptor signaling by a novel tyrosine kinase inhibitor leads to apoptosis of endothelial cells and therapy of human pancreatic carcinoma. Cancer Res 60: 2926–2935.

    CAS  PubMed  Google Scholar 

  • Canete A, Navarro S, Bermudez J, Pellin A, Castel V, Llombart-Bosch A . (2000). Angiogenesis in neuroblastoma: relationship to survival and other prognostic factors in a cohort of neuroblastoma patients. J Clin Oncol 18: 27–34.

    Article  CAS  PubMed  Google Scholar 

  • Cohen PS, Chan JP, Lipkunskaya M, Biedler JL, Seeger RC . (1994). Expression of stem cell factor and c-kit in human neuroblastoma. The Children's Cancer Group. Blood 84: 3465–3472.

    CAS  PubMed  Google Scholar 

  • Das B, Yeger H, Tsuchida R, Torkin R, Gee MF, Thorner PS et al. (2005). A hypoxia-driven vascular endothelial growth factor/Flt1 autocrine loop interacts with hypoxia-inducible factor-1alpha through mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2 pathway in neuroblastoma. Cancer Res 65: 7267–7275.

    Article  CAS  PubMed  Google Scholar 

  • Donfrancesco A, Jenkner A, Castellano A, Ilari I, Milano GM, De Sio L et al. (2004). Ifosfamide/carboplatin/etoposide (ICE) as front-line, topotecan/cyclophosphamide as second-line and oral temozolomide as third-line treatment for advanced neuroblastoma over one year of age. Acta Paediatr Suppl 93: 6–11.

    Article  CAS  PubMed  Google Scholar 

  • Eggert A, Ikegaki N, Kwiatkowski J, Zhao H, Brodeur GM, Himelstein BP . (2000). High-level expression of angiogenic factors is associated with advanced tumor stage in human neuroblastomas. Clin Cancer Res 6: 1900–1908.

    CAS  PubMed  Google Scholar 

  • Fakhari M, Pullirsch D, Paya K, Abraham D, Hofbauer R, Aharinejad S . (2002). Upregulation of vascular endothelial growth factor receptors is associated with advanced neuroblastoma. J Pediatr Surg 37: 582–587.

    Article  CAS  PubMed  Google Scholar 

  • Hanahan D, Folkman J . (1996). Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell 86: 353–364.

    Article  CAS  PubMed  Google Scholar 

  • Hishiki T, Nimura Y, Isogai E, Kondo K, Ichimiya S, Nakamura Y et al. (1998). Glial cell line-derived neurotrophic factor/neurturin-induced differentiation and its enhancement by retinoic acid in primary human neuroblastomas expressing c-Ret, GFR alpha-1, and GFR alpha-2. Cancer Res 58: 2158–2165.

    CAS  PubMed  Google Scholar 

  • Holmquist-Mengelbier L, Fredlund E, Lofstedt T, Noguera R, Navarro S, Nilsson H et al. (2006). Recruitment of HIF-1alpha and HIF-2alpha to common target genes is differentially regulated in neuroblastoma: HIF-2alpha promotes an aggressive phenotype. Cancer Cell 10: 413–423.

    Article  CAS  PubMed  Google Scholar 

  • Kaipainen A, Korhonen J, Mustonen T, van Hinsbergh VW, Fang GH, Dumont D et al. (1995). Expression of the fms-like tyrosine kinase 4 gene becomes restricted to lymphatic endothelium during development. Proc Natl Acad Sci USA 92: 3566–3570.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kane RC, Farrell AT, Madabushi R, Booth B, Chattopadhyay S, Sridhara R et al. (2009). Sorafenib for the treatment of unresectable hepatocellular carcinoma. Oncologist 14: 95–100.

    Article  CAS  PubMed  Google Scholar 

  • Kaneko M, Tsuchida Y, Mugishima H, Ohnuma N, Yamamoto K, Kawa K et al. (2002). Intensified chemotherapy increases the survival rates in patients with stage 4 neuroblastoma with MYCN amplification. J Pediatr Hematol Oncol 24: 613–621.

    Article  PubMed  Google Scholar 

  • Katzenstein HM, Salwen HR, Nguyen NN, Meitar D, Cohn SL . (2001). Antiangiogenic therapy inhibits human neuroblastoma growth. Med Pediatr Oncol 36: 190–193.

    Article  CAS  PubMed  Google Scholar 

  • Kaushal V, Mukunyadzi P, Dennis RA, Siegel ER, Johnson DE, Kohli M . (2005). Stage-specific characterization of the vascular endothelial growth factor axis in prostate cancer: expression of lymphangiogenic markers is associated with advanced-stage disease. Clin Cancer Res 11: 584–593.

    CAS  PubMed  Google Scholar 

  • Kim WY, Perera S, Zhou B, Carretero J, Yeh JJ, Heathcote SA et al. (2009). HIF2alpha cooperates with RAS to promote lung tumorigenesis in mice. J Clin Invest 119: 2160–2170.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Komuro H, Kaneko S, Kaneko M, Nakanishi Y . (2001). Expression of angiogenic factors and tumor progression in human neuroblastoma. J Cancer Res Clin Oncol 127: 739–743.

    CAS  PubMed  Google Scholar 

  • Kushner BH, Kramer K, Meyers PA, Wollner N, Cheung NK . (2000). Pilot study of topotecan and high-dose cyclophosphamide for resistant pediatric solid tumors. Med Pediatr Oncol 35: 468–474.

    Article  CAS  PubMed  Google Scholar 

  • Ladenstein R, Philip T, Lasset C, Hartmann O, Garaventa A, Pinkerton R et al. (1998). Multivariate analysis of risk factors in stage 4 neuroblastoma patients over the age of one year treated with megatherapy and stem-cell transplantation: a report from the European Bone Marrow Transplantation Solid Tumor Registry. J Clin Oncol 16: 953–965.

    Article  CAS  PubMed  Google Scholar 

  • Langer I, Vertongen P, Perret J, Fontaine J, Atassi G, Robberecht P . (2000). Expression of vascular endothelial growth factor (VEGF) and VEGF receptors in human neuroblastomas. Med Pediatr Oncol 34: 386–393.

    Article  CAS  PubMed  Google Scholar 

  • Lesslie DP, Summy JM, Parikh NU, Fan F, Trevino JG, Sawyer TK et al. (2006). Vascular endothelial growth factor receptor-1 mediates migration of human colorectal carcinoma cells by activation of Src family kinases. Br J Cancer 94: 1710–1717.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Maris JM, Courtright J, Houghton PJ, Morton CL, Gorlick R, Kolb EA et al. (2008). Initial testing of the VEGFR inhibitor AZD2171 by the pediatric preclinical testing program. Pediatr Blood Cancer 50: 581–587.

    Article  PubMed  Google Scholar 

  • Maris JM, Hogarty MD, Bagatell R, Cohn SL . (2007). Neuroblastoma. Lancet 369: 2106–2120.

    Article  CAS  PubMed  Google Scholar 

  • Matthay KK, Villablanca JG, Seeger RC, Stram DO, Harris RE, Ramsay NK et al. (1999). Treatment of high-risk neuroblastoma with intensive chemotherapy, radiotherapy, autologous bone marrow transplantation, and 13-cis-retinoic acid.Children's Cancer Group. N Engl J Med 341: 1165–1173.

    Article  CAS  PubMed  Google Scholar 

  • Meister B, Grunebach F, Bautz F, Brugger W, Fink FM, Kanz L et al. (1999). Expression of vascular endothelial growth factor (VEGF) and its receptors in human neuroblastoma. Eur J Cancer 35: 445–449.

    Article  CAS  PubMed  Google Scholar 

  • Meitar D, Crawford SE, Rademaker AW, Cohn SL . (1996). Tumor angiogenesis correlates with metastatic disease, N-myc amplification, and poor outcome in human neuroblastoma. J Clin Oncol 14: 405–414.

    Article  CAS  PubMed  Google Scholar 

  • Nakamura T, Ishizaka Y, Nagao M, Hara M, Ishikawa T . (1994). Expression of the ret proto-oncogene product in human normal and neoplastic tissues of neural crest origin. J Pathol 172: 255–260.

    Article  CAS  PubMed  Google Scholar 

  • O'Farrell AM, Abrams TJ, Yuen HA, Ngai TJ, Louie SG, Yee KW et al. (2003). SU11248 is a novel FLT3 tyrosine kinase inhibitor with potent activity in vitro and in vivo. Blood 101: 3597–3605.

    Article  CAS  PubMed  Google Scholar 

  • Peddinti R, Zeine R, Luca D, Seshadri R, Chlenski A, Cole K et al. (2007). Prominent microvascular proliferation in clinically aggressive neuroblastoma. Clin Cancer Res 13: 3499–3506.

    Article  CAS  PubMed  Google Scholar 

  • Peng XH, Karna P, Cao Z, Jiang BH, Zhou M, Yang L . (2006). Cross-talk between epidermal growth factor receptor and hypoxia-inducible factor-1alpha signal pathways increases resistance to apoptosis by up-regulating survivin gene expression. J Biol Chem 281: 25903–25914.

    Article  CAS  PubMed  Google Scholar 

  • Pennacchietti S, Michieli P, Galluzzo M, Mazzone M, Giordano S, Comoglio PM . (2003). Hypoxia promotes invasive growth by transcriptional activation of the met protooncogene. Cancer Cell 3: 347–361.

    Article  PubMed  Google Scholar 

  • Phillips RJ, Mestas J, Gharaee-Kermani M, Burdick MD, Sica A, Belperio JA et al. (2005). Epidermal growth factor and hypoxia-induced expression of CXC chemokine receptor 4 on non-small cell lung cancer cells is regulated by the phosphatidylinositol 3-kinase/PTEN/AKT/mammalian target of rapamycin signaling pathway and activation of hypoxia inducible factor-1alpha. J Biol Chem 280: 22473–22481.

    Article  CAS  PubMed  Google Scholar 

  • Pickering LM, Pyle L, Larkin JM . (2009). Sunitinib is superior to interferon alpha with respect to quality of life for patients with renal cell carcinoma. Nat Clin Pract Oncol 6: 6–7.

    Article  CAS  PubMed  Google Scholar 

  • Richardson DS, Lai AZ, Mulligan LM . (2006). RET ligand-induced internalization and its consequences for downstream signaling. Oncogene 25: 3206–3211.

    Article  CAS  PubMed  Google Scholar 

  • Sandler A, Gray R, Perry MC, Brahmer J, Schiller JH, Dowlati A et al. (2006). Paclitaxel-carboplatin alone or with bevacizumab for non-small-cell lung cancer. N Engl J Med 355: 2542–2550.

    Article  CAS  PubMed  Google Scholar 

  • Semenza GL . (2003). Targeting HIF-1 for cancer therapy. Nat Rev Cancer 3: 721–732.

    Article  CAS  PubMed  Google Scholar 

  • Shusterman S, Maris JM . (2005). Prospects for therapeutic inhibition of neuroblastoma angiogenesis. Cancer Lett 228: 171–179.

    Article  CAS  PubMed  Google Scholar 

  • Stommel JM, Kimmelman AC, Ying H, Nabioullin R, Ponugoti AH, Wiedemeyer R et al. (2007). Coactivation of receptor tyrosine kinases affects the response of tumor cells to targeted therapies. Science 318: 287–290.

    Article  CAS  PubMed  Google Scholar 

  • Wey JS, Fan F, Gray MJ, Bauer TW, McCarty MF, Somcio R et al. (2005). Vascular endothelial growth factor receptor-1 promotes migration and invasion in pancreatic carcinoma cell lines. Cancer 104: 427–438.

    Article  CAS  PubMed  Google Scholar 

  • Wilhelm SM, Carter C, Tang L, Wilkie D, McNabola A, Rong H et al. (2004). BAY 43-9006 exhibits broad spectrum oral antitumor activity and targets the RAF/MEK/ERK pathway and receptor tyrosine kinases involved in tumor progression and angiogenesis. Cancer Res 64: 7099–7109.

    Article  CAS  PubMed  Google Scholar 

  • Witte D, Thomas A, Ali N, Carlson N, Younes M . (2002). Expression of the vascular endothelial growth factor receptor-3 (VEGFR-3) and its ligand VEGF-C in human colorectal adenocarcinoma. Anticancer Res 22: 1463–1466.

    CAS  PubMed  Google Scholar 

  • Yamagata K, Tagami M, Ikeda K, Tsumagari S, Yamori Y, Nara Y . (2002). Differential regulation of glial cell line-derived neurotrophic factor (GDNF) mRNA expression during hypoxia and reoxygenation in astrocytes isolated from stroke-prone spontaneously hypertensive rats. Glia 37: 1–7.

    Article  PubMed  Google Scholar 

  • Yang AD, Camp ER, Fan F, Shen L, Gray MJ, Liu W et al. (2006). Vascular endothelial growth factor receptor-1 activation mediates epithelial to mesenchymal transition in human pancreatic carcinoma cells. Cancer Res 66: 46–51.

    Article  CAS  PubMed  Google Scholar 

  • Yang JC . (2004). Bevacizumab for patients with metastatic renal cancer: an update. Clin Cancer Res 10: 6367S–6370S.

    Article  CAS  PubMed  Google Scholar 

  • Yoshida D, Kim K, Noha M, Teramoto A . (2006). Hypoxia inducible factor 1-alpha regulates of platelet derived growth factor-B in human glioblastoma cells. J Neurooncol 76: 13–21.

    Article  CAS  PubMed  Google Scholar 

  • Zhang L, Smith KM, Chong AL, Stempak D, Yeger H, Marrano P et al. (2009). in vivo antitumor and antimetastatic activity of sunitinib in preclinical neuroblastoma mouse model. Neoplasia 11: 426–435.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhong H, Chiles K, Feldser D, Laughner E, Hanrahan C, Georgescu MM et al. (2000). Modulation of hypoxia-inducible factor 1alpha expression by the epidermal growth factor/phosphatidylinositol 3-kinase/PTEN/AKT/FRAP pathway in human prostate cancer cells: implications for tumor angiogenesis and therapeutics. Cancer Res 60: 1541–1545.

    CAS  PubMed  Google Scholar 

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Acknowledgements

JVH is a Damon Runyon-Lilly Clinical Investigator supported in part by the Damon Runyon Cancer Research Foundation (CI 24-04) and the MD Anderson Cancer Center Physician Scientist Award. This work was also supported in part by the Lorrie Olivier Neuroblastoma Research Fund.

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Authors and Affiliations

  1. Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    M B Nilsson, L Xu, T Cascone, H K Wu, B Saigal & J V Heymach

  2. Division of Pediatrics, Children's Cancer Hospital, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    P E Zage, L Zeng & P A Zweidler-McKay

  3. Department of Cancer Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    J V Heymach

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JVH receives research support and serves on the advisory board for Pfizer.

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Nilsson, M., Zage, P., Zeng, L. et al. Multiple receptor tyrosine kinases regulate HIF-1α and HIF-2α in normoxia and hypoxia in neuroblastoma: implications for antiangiogenic mechanisms of multikinase inhibitors. Oncogene 29, 2938–2949 (2010). https://doi.org/10.1038/onc.2010.60

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