Abstract PR04: Deciphering radiotherapy resistance mechanisms in HPV negative and positive head and neck cancers
Authors: Tavassoli Mahvash 
Published: September 2023

Abstract
Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide. The prognosis of HNSCC is poor with a 5-year survival rate for all stages around 55%. HNSCC can be classified into two subtypes, human papilloma virus (HPV)-negative, caused mainly by tobacco use, and HPV-positive subtype. Radiotherapy is one of the cornerstones of the treatment of HNSCC. However, radioresistance is common and is associated with a high risk of recurrence. Radioresistance may be pre-existing, or acquired, and may be direct (tumor-cell autonomous) or indirect, mediated by RT-mediated changes to the TME such as cellular and/or extracellular matrix (ECM) changes to the stroma. These pathways are not fully understood. Better understanding of the intrinsic as well as RT-induced resistance is needed to improve survival of hard-to-treat HNSCC patients. HPV-positive HNSCC have significantly better RT response and prognosis, however currently they are treated with similarly high dose RT which causes significant adverse side-effects. As the incidence of HPV-induced HNSCC is increasing very fast, the current focus is on treatment de-escalation to reduce toxicity without compromising outcome. EGFR upregulation is an established biomarker of treatment resistance and aggressiveness in head HNSCC. EGFR-targeted therapies have shown benefits for HPV-negative HNSCC; surprisingly, inhibiting EGFR in HPV-associated HNSCC led to inferior therapeutic outcomes suggesting opposing roles for EGFR in the two HNSCC subtypes. We have recently shown that EGFR activation regulates the DNA damage response pathway differently in the two HNSCC subtype. Here, we further investigated the link between EGFR with HPV-infected HNSCC particularly the regulation of HPV oncoproteins E6 and E7. We demonstrate that EGFR overexpression suppresses cellular proliferation and increases radiosensitivity of HPV-positive HNSCC cell lines. EGFR overexpression was shown to inhibit protein expression of BRD4, a known cellular transcriptional regulator of HPV E6/E7 expression and DNA damage repair facilitator. Using in vitro 2D and 3D spheroid models of HNSCC we showed that EGFR inhibition by cetuximab restored the expression of BRD4 leading to increased HPV E6 and E7 transcription. Concordantly, pharmacological inhibition of BRD4 led to suppression of HPV E6 and E7 transcription, delayed cellular proliferation and sensitised HPV-positive HNSCC cells to ionising radiation. This effect was shown to be mediated through EGFR-induced upregulation of microRNA-9-5p and consequent silencing of its target BRD4 at protein translational level, repressing HPV E6 and E7 transcription and restoring p53 tumor suppressor functions. These results suggest a novel mechanism for EGFR inhibition of HPV E6/E7 oncoprotein expression through an epigenetic pathway, mediated through microRNA-9-5p/BRD4 regulation. The results may help better future design of radiotherapy combination treatment of HPV-positive and negative HNSCC, such as combination with BRD4 inhibitors to improve HNSCC therapeutic outcome.
Citation Format: Mahvash Tavassoli. Deciphering radiotherapy resistance mechanisms in HPV negative and positive head and neck cancers [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Innovating through Basic, Clinical, and Translational Research; 2023 Jul 7-8; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2023;29(18_Suppl):Abstract nr PR04.