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EGFRvIII gene rearrangement is an early event in glioblastoma tumorigenesis and expression defines a hierarchy modulated by epigenetic mechanisms

Oncogene volume 32, pages 2670–2681 (2013)Cite this article

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Abstract

Amplification and rearrangements of the epidermal growth factor receptor (EGFR) gene are frequently found in glioblastoma multiforme (GBM). The most common variant is EGFR variant III (EGFRvIII). Research suggests that EGFRvIII could be a marker for a cancer stem cell or tumor-initiating population. If amplification and rearrangement are early events in tumorigenesis, this implies that they should be preserved throughout the tumor. However, in primary GBM, EGFRvIII expression is focal and sporadic. Unexpectedly, we found EGFR amplification and rearrangement throughout the tumor, including regions with no EGFRvIII expression, suggesting that mechanisms exist to modulate EGFRvIII expression even in the presence of high gene amplification. To study this phenomenon, we characterized three GBM cell lines with endogenous EGFRvIII. EGFRvIII expression was heterogeneous, with both positive and negative populations maintaining the genetic alterations, akin to primary tumors. Furthermore, EGFRvIII defined a hierarchy where EGFRvIII-positive cells gave rise to additional positive and negative cells. Only cells that had recently lost EGFRvIII expression could re-express EGFRvIII, providing an important buffer for maintaining EGFRvIII-positive cell numbers. Epigenetic mechanisms had a role in maintaining heterogeneous EGFRvIII expression. Demethylation induced a 20–60% increase in the percentage of EGFRvIII-positive cells, indicating that some cells could re-express EGFRvIII. Surprisingly, inhibition of histone deacetylation resulted in a 50–80% reduction in EGFRvIII expression. Collectively, this data demonstrates that EGFR amplification and rearrangement are early events in tumorigenesis and EGFRvIII follows a model of hierarchical expression. Furthermore, EGFRvIII expression is restricted by epigenetic mechanisms, suggesting that drugs that modulate the epigenome might be used successfully in glioblastoma tumors.

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Acknowledgements

We thank Dr Neethan Lobo, Dr Siddhartha Mitra, Dr A Hunter Shain and the Stanford FACS facility for technical advice. Additionally, we thank Dana Bangs and Dr Athena Cherry at the Stanford Cytogenetics Laboratory for their assistance with FISH staining and analysis. This work was supported by the National Defense Science and Engineering Graduate Research Fellowship, the National Science Foundation Graduate Research Fellowship, NIH Grants CA124832, RC2 CA14891, the Lucille Packard Children’s Foundation and a research grant from the National Brain Tumor Foundation.

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Authors and Affiliations

  1. Department of Neurosurgery, Stanford University School of Medicine, Stanford, CA, USA

    C A Del Vecchio & A J Wong

  2. Cancer Biology Program, Stanford University School of Medicine, Stanford, CA, USA

    C A Del Vecchio, C P Giacomini, J R Pollack & A J Wong

  3. Department of Pathology, Stanford University Medical Center, Stanford, CA, USA

    C P Giacomini, H Vogel, K C Jensen & J R Pollack

  4. Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA

    K C Jensen

  5. Department of Oncology, Biology and Genetics, University of Genova, Genova, Italy

    T Florio

  6. Department of Clinical and Biological Sciences, Laboratory of Molecular Neuro-Oncology, University of Basel, Basel, Switzerland

    A Merlo

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Correspondence to A J Wong.

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Del Vecchio, C., Giacomini, C., Vogel, H. et al. EGFRvIII gene rearrangement is an early event in glioblastoma tumorigenesis and expression defines a hierarchy modulated by epigenetic mechanisms. Oncogene 32, 2670–2681 (2013). https://doi.org/10.1038/onc.2012.280

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