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Heat shock protein 90 (HSP90) is overexpressed in p16-negative oropharyngeal squamous cell carcinoma, and its inhibition in vitro potentiates the effects of chemoradiation

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Abstract

Purpose

Cisplatin and radiation therapy remain the current standard for treating locally advanced SCCHN. Novel treatment approaches are needed, especially in patients with human papilloma virus (HPV)-negative disease who have worse outcomes despite multimodality therapy.

Methods

Using our institutional review board approved database, we obtained twenty oropharyngeal squamous cell carcinoma (SCC) tissue samples: ten p16 positive, ten p16-negative. Because p16 expression is strongly associated with HPV positivity in oropharyngeal SCC, p16 status was used as a marker of HPV. We subsequently analyzed, via immunohistochemistry, heat shock protein 90 (HSP90) protein levels. Using HPV-positive and HPV-negative SCC cell lines, we compared baseline HSP90 expression levels and the effect of the HSP90 inhibitor ganetespib on viability and apoptosis. Clonogenic survival of HPV-negative cells treated with ganetespib, radiation therapy, and/or cisplatin was then investigated. We characterize the effects of ganetespib on proteins that are thought to drive DNA damage resistance in HPV-negative cells.

Results

HSP90 expression was significantly higher in p16-negative compared with p16-positive samples (p = 0.016) and in HPV-negative cell lines compared with positive cells. Ganetespib increased cytotoxicity and induced apoptosis in HPV-negative more than positive cells. Adding ganetespib to cisplatin and/or radiation therapy in HPV-negative cells further decreased clonogenic survival. Finally, ganetespib downregulated expressions of EGFR, ERK, AKT, p53, and HIF-1α.

Conclusions

Ganetespib inhibited HPV-negative SCCHN viability and potentiated cell kill when combined with cisplatin or radiation therapy in vitro. With HSP90 expression higher in HPV-negative cells and in p16-negative patients, further exploration of the clinical activity of HSP90 inhibitors in SCCHN is warranted.

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Conflict of interest

All authors declare that they have neither financial nor non-financial completing interests.

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Correspondence to Kirtesh Patel.

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280_2014_2584_MOESM1_ESM.docx

Client proteins/pathways are prognostic or predictive biomarkers specifically for HPV-negative SCCHN. EGFR and mutant p53 have been established as client HSP90 proteins. HIF-1α, a marker of hypoxia, is also a known client protein of HSP90. Above table demonstrates EGFR can be targeted by cetuximab, but only patients with HPV+/p16+ characteristics tended to predict response. Mutant p53 is a prognostic biomarker in SCC of the head and neck of patients who are likely HPV negative given that HPV-positive patients express E7 which suppresses p53 expression. Hypoxic inhibitors sensitized HPV-negative, but not HPV-positive patients to concurrent cisplatin and radiation therapy. The efficacy of targeting these important client proteins is demonstrated in Fig. 5. [LC—local control; OS—overall survival]. (DOCX 21 kb)

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Patel, K., Wen, J., Magliocca, K. et al. Heat shock protein 90 (HSP90) is overexpressed in p16-negative oropharyngeal squamous cell carcinoma, and its inhibition in vitro potentiates the effects of chemoradiation. Cancer Chemother Pharmacol 74, 1015–1022 (2014). https://doi.org/10.1007/s00280-014-2584-8

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  • DOI: https://doi.org/10.1007/s00280-014-2584-8

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