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EGFRvIII promotes glioma angiogenesis and growth through the NF-κB, interleukin-8 pathway

Oncogene volume 31, pages 4054–4066 (2012)Cite this article

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Abstract

Sustaining a high growth rate requires tumors to exploit resources in their microenvironment. One example of this is the extensive angiogenesis that is a typical feature of high-grade gliomas. Here, we show that expression of the constitutively active mutant epidermal growth factor receptor, ΔEGFR (EGFRvIII, EGFR*, de2-7EGFR) is associated with significantly higher expression levels of the pro-angiogenic factor interleukin (IL)-8 in human glioma specimens and glioma stem cells. Furthermore, the ectopic expression of ΔEGFR in different glioma cell lines caused up to 60-fold increases in the secretion of IL-8. Xenografts of these cells exhibit increased neovascularization, which is not elicited by cells overexpressing wild-type (wt)EGFR or ΔEGFR with an additional kinase domain mutation. Analysis of the regulation of IL-8 by site-directed mutagenesis of its promoter showed that ΔEGFR regulates its expression through the transcription factors nuclear factor (NF)-κB, activator protein 1 (AP-1) and CCAAT/enhancer binding protein (C/EBP). Glioma cells overexpressing ΔEGFR showed constitutive activation and DNA binding of NF-κB, overexpression of c-Jun and activation of its upstream kinase c-Jun N-terminal kinase (JNK) and overexpression of C/EBPβ. Selective pharmacological or genetic targeting of the NF-κB or AP-1 pathways efficiently blocked promoter activity and secretion of IL-8. Moreover, RNA interference-mediated knock-down of either IL-8 or the NF-κB subunit p65, in ΔEGFR-expressing cells attenuated their ability to form tumors and to induce angiogenesis when injected subcutaneously into nude mice. On the contrary, the overexpression of IL-8 in glioma cells lacking ΔEGFR potently enhanced their tumorigenicity and produced highly vascularized tumors, suggesting the importance of this cytokine and its transcription regulators in promoting glioma angiogenesis and tumor growth.

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Acknowledgements

We thank Donna Harclerode and Donald Pizzo for technical assistance with immunostaining and Dr Cameron Brennan for providing the GBM tumorspheres. This work was supported by an award from the Goldhirsh Foundation (to FBF), and NIH Grant P01-CA95616 (to WKC, FBF), as well as NIH RO1CA130966, a grant with Pennsylvania Department of Health and an Innovative Research Scholar Award of the Hillman Foundation (to SYC). WKC is a fellow of the National Foundation for Cancer Research. MM Inda thanks the Gobierno de Navarra, Spain, and the American Brain Tumor Association in Honor of Walter Terlik for the fellowships received. R Bonavia was supported by a fellowship from Federazione Italiana Ricerca Cancro.

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  1. R Bonavia and M M Inda: These authors contributed equally to this work.

Authors and Affiliations

  1. Ludwig Institute for Cancer Research, University of California, La Jolla, CA, USA

    R Bonavia, M M Inda, W K Cavenee & F B Furnari

  2. Department of Pathology, School of Medicine, University of California, La Jolla, CA, USA

    R Bonavia & S Vandenberg

  3. Cancer Center, University of California, La Jolla, CA, USA

    S Vandenberg, W K Cavenee & F B Furnari

  4. Cancer Institute, University of Pittsburgh, Pittsburgh, PA, USA

    S-Y Cheng

  5. Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA

    S-Y Cheng

  6. Department of Neurosurgery, Kyorin University Faculty of Medicine, Tokyo, Japan

    M Nagane

  7. Alnylam Europe AG, Kulmbach, Germany

    P Hadwiger & P Tan

  8. Alnylam Pharmaceuticals, Inc., Cambridge, MA, USA

    D W Y Sah

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  1. R Bonavia
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Correspondence to F B Furnari.

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Bonavia, R., Inda, M., Vandenberg, S. et al. EGFRvIII promotes glioma angiogenesis and growth through the NF-κB, interleukin-8 pathway. Oncogene 31, 4054–4066 (2012). https://doi.org/10.1038/onc.2011.563

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