EGFRvIII and DNA double-strand break repair: A molecular mechanism for radioresistance in glioblastoma

Bipasha Mukherjee, Brian McEllin, Cristel V. Camacho, Nozomi Tomimatsu, Shyam Sirasanagandala, Suraj Nannepaga, Kimmo J. Hatanpaa, Bruce Mickey, Christopher Madden, Elizabeth Maher, David A. Boothman, Frank Furnari, Webster K. Cavenee, Robert M. Bachoo, Sandeep Burma

Research output: Contribution to journalArticle

174 Citations (Scopus)

Abstract

Glioblastoma multiforme (GBM) is the most lethal of brain tumors and is highly resistant to ionizing radiation (IR) and chemotherapy. Here, we report on a molecular mechanism by which a key glioma-specific mutation, epidermal growth factor receptor variant III (EGFRvIII), confers radiation resistance. Using Ink4a/Arf-deficient primary mouse astrocytes, primary astrocytes immortalized by p53/Rb suppression, as well as human U87 glioma cells, we show that EGFRvIII expression enhances clonogenic survival following IR. This enhanced radioresistance is due to accelerated repair of DNA double-strand breaks (DSB), the most lethal lesion inflicted by IR. The EGFR inhibitor gefitinib (Iressa) and the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 attenuate the rate of DSB repair. Importantly, expression of constitutively active, myristylated Akt-1 accelerates repair, implicating the PI3K/Akt-1 pathway in radioresistance. Most notably, EGFRvIII-expressing U87 glioma cells show elevated activation of a key DSB repair enzyme, DNA-dependent protein kinase catalytic subunit (DNA-PKcs). Enhanced radioresistance is abrogated by the DNA-PKcs-specific inhibitor NU7026, and EGFRvIII fails to confer radioresistance in DNA-PKcs-deficient cells. In vivo, orthotopic U87-EGFRvIII-derived tumors display faster rates of DSB repair following whole-brain radiotherapy compared with U87-derived tumors. Consequently, EGFRvIII-expressing tumors are radioresistant and continue to grow following whole-brain radiotherapy with little effect on overall survival. These in vitro and in vivo data support our hypothesis that EGFRvIII expression promotes DNA-PKcs activation and DSB repair, perhaps as a consequence of hyperactivated PI3K/Akt-1 signaling. Taken together, our results raise the possibility that EGFR and/or DNA-PKcs inhibition concurrent with radiation may be an effective therapeutic strategy for radiosensitizing high-grade gliomas.

Original languageEnglish (US)
Pages (from-to)4252-4259
Number of pages8
JournalCancer Research
Volume69
Issue number10
DOIs
StatePublished - May 15 2009

Fingerprint

Double-Stranded DNA Breaks
Glioblastoma
DNA-Activated Protein Kinase
Epidermal Growth Factor Receptor
Phosphatidylinositol 3-Kinase
Catalytic Domain
Glioma
Ionizing Radiation
Astrocytes
Radiotherapy
Radiation
Neoplasms
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Survival
Brain
Brain Neoplasms
Drug Therapy
Mutation
Enzymes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Mukherjee, B., McEllin, B., Camacho, C. V., Tomimatsu, N., Sirasanagandala, S., Nannepaga, S., ... Burma, S. (2009). EGFRvIII and DNA double-strand break repair: A molecular mechanism for radioresistance in glioblastoma. Cancer Research, 69(10), 4252-4259. https://doi.org/10.1158/0008-5472.CAN-08-4853

EGFRvIII and DNA double-strand break repair : A molecular mechanism for radioresistance in glioblastoma. / Mukherjee, Bipasha; McEllin, Brian; Camacho, Cristel V.; Tomimatsu, Nozomi; Sirasanagandala, Shyam; Nannepaga, Suraj; Hatanpaa, Kimmo J.; Mickey, Bruce; Madden, Christopher; Maher, Elizabeth; Boothman, David A.; Furnari, Frank; Cavenee, Webster K.; Bachoo, Robert M.; Burma, Sandeep.

In: Cancer Research, Vol. 69, No. 10, 15.05.2009, p. 4252-4259.

Research output: Contribution to journalArticle

Mukherjee, B, McEllin, B, Camacho, CV, Tomimatsu, N, Sirasanagandala, S, Nannepaga, S, Hatanpaa, KJ, Mickey, B, Madden, C, Maher, E, Boothman, DA, Furnari, F, Cavenee, WK, Bachoo, RM & Burma, S 2009, 'EGFRvIII and DNA double-strand break repair: A molecular mechanism for radioresistance in glioblastoma', Cancer Research, vol. 69, no. 10, pp. 4252-4259. https://doi.org/10.1158/0008-5472.CAN-08-4853
Mukherjee B, McEllin B, Camacho CV, Tomimatsu N, Sirasanagandala S, Nannepaga S et al. EGFRvIII and DNA double-strand break repair: A molecular mechanism for radioresistance in glioblastoma. Cancer Research. 2009 May 15;69(10):4252-4259. https://doi.org/10.1158/0008-5472.CAN-08-4853
Mukherjee, Bipasha ; McEllin, Brian ; Camacho, Cristel V. ; Tomimatsu, Nozomi ; Sirasanagandala, Shyam ; Nannepaga, Suraj ; Hatanpaa, Kimmo J. ; Mickey, Bruce ; Madden, Christopher ; Maher, Elizabeth ; Boothman, David A. ; Furnari, Frank ; Cavenee, Webster K. ; Bachoo, Robert M. ; Burma, Sandeep. / EGFRvIII and DNA double-strand break repair : A molecular mechanism for radioresistance in glioblastoma. In: Cancer Research. 2009 ; Vol. 69, No. 10. pp. 4252-4259.
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