Targeting Viral DNA and Promoter Hypermethylation in Salivary Rinses for Recurrent HPV‐Positive Oropharyngeal Cancer

Abstract

Objective The incidence and survivorship of human papillomavirus (HPV)–associated oropharyngeal squamous cell carcinoma (OPSCC) are increasing. Presence of HPV DNA and epigenetic alterations in salivary rinses are independently associated with clinical prognosis. We evaluated the utility of a combined panel in detecting disease recurrence during surveillance. We also assessed the assay’s applicability in screening for HPV+ OPSCC. Study Design Retrospective cohort study. Setting Two tertiary academic hospitals. Subjects and Methods Forty-nine patients with posttreatment OPSCC were enrolled. Separately, 21 treatment-naive patients and 40 controls were included in the screening analysis. Salivary rinses were obtained from these cohorts and biomarker levels were quantified. Receiver operative characteristic (ROC) curves and multivariate logistic models were used to assess performance of biomarker combinations. Results Eight patients (16.3%) in the posttreatment cohort developed locoregional recurrence. Recurrence was associated with alcohol use (odds ratio [OR], 6.12; 95% confidence interval [CI], 0.26-3.79) and advanced nodal disease (OR, 2.21; 95% CI, 1.52-3.01). A panel of HPV DNA and methylated EDNRB improved detection of recurrent disease (area under the curve [AUC], 0.88) compared to single markers (AUC, 0.69-0.78). Positive biomarkers preceded clinical detection by 2.4 ± 1.6 months and was associated with nearly 40-fold risk of recurrence (OR, 36.4; 95% CI, 1.15-45.22). Within the screening analysis, single biomarkers demonstrated moderate sensitivity and specificity (AUC, 0.59-0.83) in the detection of primary disease. A panel combining HPV DNA markers with methylated EDNRB and methylated PAX5 improved AUC to 0.93. Conclusion Detection of high-risk HPV DNA or aberrant hypermethylation in oral rinses is associated with presence and recurrence of OPSCC. Targeting both markers in saliva may have utility in long-term surveillance.

Salivary DNA Methylation as an Epigenetic Biomarker for Head and Neck Cancer. Part I: A Diagnostic Accuracy Meta-Analysis

“…After title and abstract review, 27 articles were submitted for full-text reading, of which nine were excluded for the following reasons: non-independent cancer group (two articles); reviews, letters, personal opinions, book chapters, case reports, conference abstracts, and meetings (three articles); absence of a healthy control group (one article); saliva enriched with brush oral cytology (two articles); and insufficient information for meta-analysis (one article). In the end, 18 articles met the inclusion criteria for final analysis [ 10 , 11 , 12 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 ].…”

“…A total of 34 different genes were identified in the studies. Five studies evaluated the methylation status of a single gene [ 31 , 34 , 36 , 39 , 41 ] and 13 studies evaluated two or more genes [ 10 , 11 , 12 , 30 , 32 , 33 , 35 , 37 , 38 , 40 , 42 , 43 , 44 ]. Ten studies focused on gene promoter methylation panels combining two to four genes, whereas eight studies evaluated only single genes.…”

Current salivary biomarkers for detection of human papilloma virus‐induced oropharyngeal squamous cell carcinoma

“…When screening for primary disease, a combination of any HPV biomarker plus any methylated DNA region resulted in a sensitivity of 100% and a specificity of 85%. Specific combinations also proved to be very impressive, including, among others, a panel consisting of BR E1‐5, HR E5L2‐4, EDNRB, and hypermethylated PAX5 that achieved a sensitivity of 97% and specificity of 87% 57 . Ideally, results would be verified by future study with a larger cohort size.…”

“…Specific combinations also proved to be very impressive, including, among others, a panel consisting of BR E1-5, HR E5L2-4, EDNRB, and hypermethylated PAX5 that achieved a sensitivity of 97% and specificity of 87%. 57 Ideally, results would be verified by future study with a larger cohort size.…”

Current Status of Human Papillomavirus-Related Head and Neck Cancer: From Viral Genome to Patient Care

“…However, their role in the oncogenesis of HNSCC remains elusive. In conclusion, epigenetic alterations might be useful in identifying subgroups of tumors, predicting clinical outcomes and providing potential targets (Kostareli et al 2013(Kostareli et al , 2016Ren et al 2018;Shen et al 2020).…”