The Use of HPV16-E5, EGFR, and pEGFR as Prognostic Biomarkers for Oropharyngeal Cancer Patients

Abstract

Background: Anti-epidermal-growth-factor-receptor (EGFR) therapies in combination with radiotherapy are being studied on de-escalation clinical trials for HPV-related oropharyngeal cancer (OPC) patients. The HPV16-E5 oncoprotein increases recycling of activated EGFR to the cell surface, enhancing factor signal transduction. Our aim was to evaluate viral HPV16-E5 oncogene expression as well as EGFR and phosphorylated-EGFR (pEGFR), protein levels as biomarkers for clinical outcome in a retrospective cohort of OPC patients.Methods: Formalin-fixed-paraffin-embedded OPCs were collected from 1990 to 2013. OPC samples containing HPV-DNA were subject to viral E6*I mRNA detection and p16INK4a immunohistochemistry (IHC). HPV16-positive cases were evaluated for HPV16-E5 (RT-PCR) and EGFR/pEGFR (IHC). A stratified and matched random sample of HPV-negative samples was used as control and evaluated for EGFR/pEGFR. Overall survival (OS) and disease free survival (DFS) estimates were assessed for locally advanced OPC patients (stage III, IVa,b 7th edition).Results: Among 788 OPC patient samples, 53 were double positive for HPV16-DNA/p16INK4a. HPV16-E5 expression was found in 41 of 53 samples (77.4%). EGFR expression was observed in 37.7 vs 70.8% of HPV16-positive vs HPV-negative samples, respectively; (adjusted OR = 0.15) 5% CI = 0.04–0.56]). Expression of pEGFR followed an inverse pattern with 39.6 and 24.9% detection in HPV16-positive and HPV-negative samples; (adjusted OR = 1.58 [95% CI = 0.48–5.17]). Within HPV16-positive cases, no association between HPV16-E5/EGFR nor pEGFR was observed. With a median follow-up of 39.36 months (min = 0.03 – max = 272.07), the combination of HPV status and EGFR or pEGFR expression were predictors of better OS (p < 0.001, for both) and DFS (p < 0.001 for EGFR and p = 0.003 for pEGFR).Conclusions: HPV16-E5 is highly expressed on HPV16-positive OPCs. Interestingly, HPV16-positive cases expressed significantly more pEGFR while HPV-negative cases expressed more EGFR. The combinations of HPV status and EGFR or pEGFR may be useful biomarkers for evaluating prognosis outcome in OPC patients.

Prevalence of Human Papillomavirus (HPV) Infection and the Association with Survival in Saudi Patients with Head and Neck Squamous Cell Carcinoma

“…Nevertheless, our results are in overall agreement with published data with the overwhelming belief that p16-positive HNSCC have improved locoregional tumor control and survival with conventional therapy [21,27,41]. Potential future refinement could be brought about by including the copy number variation of the CDKN2A gene that encodes p16ink4a [42], and involving other related prognostic biomarkers such as epidermal growth factor receptor (EGFR), and key transcription factors as molecular signature of HPV presence [19,43], especially for de-escalation of radiotherapy combined with anti-EGFR receptor treatment [44].…”

The Not-So-Good, the Bad and the Ugly: HPV E5, E6 and E7 Oncoproteins in the Orchestration of Carcinogenesis

“…In addition, studies in transgenic mice suggested that the expression of HPV-16 E5 in stratified squamous epithelia led to a higher frequency of spontaneous skin tumours [ 109 ] and caused cervical cancer when a prolonged oestrogen treatment was given [ 109 , 110 ]. As discussed earlier, upon integration of the HPV genome in high-risk HPV-18, E5 is often disrupted; however, in the case of HPV-16-positive cervical and oropharyngeal cancers, the expression of E5 is more likely and has been shown to be detectable [ 111 , 112 , 113 , 114 , 115 ], suggesting that E5 may contribute to malignant progression of the cancer.…”

Development of HPV16,18,31,45 E5 and E7 peptides-based vaccines predicted by immunoinformatics tools

“…On the other hand, the presence of E5 viral transcripts could be a major marker of active viral infection, and subsequently a target of immunotherapy. HPV16 E5 was highly expressed in HPV16-positive oropharyngeal cancer (OPC) patients (Taberna et al 2018). Generally, most studies have focused on E7 oncoprotein, because it is more abundantly expressed and better characterized immunologically.…”