HPV-driven Oropharyngeal Cancer Research Articles

Oral testing for high-risk human papillomavirus DNA and E6/E7 messenger RNA in healthy individuals at risk for oral infection.

Testing for oral high-risk human papillomavirus (HPV) DNA may be useful for identifying individuals at increased risk for HPV-driven oropharyngeal cancer (OPC). However, positivity for HPV DNA provides no information on the transforming potential of the infection. In contrast, the detection of high-risk HPV E6/E7 messenger RNA (mRNA) may help to identify clinically significant infections because of the indispensable role of E6/E7 viral oncoproteins in the carcinogenic process. Oral rinses were collected with a mouthwash from cancer-free individuals at increased risk for oral HPV infection. High-risk HPV DNA and mRNA were evaluated via the testing of the oral rinses with the Linear Array HPV genotyping test and the Aptima HPV assay, respectively. Overall, 310 subjects with no clinical evidence of lesions of the oral cavity and oropharynx were included in the study. Thirty-three (10.6%) harbored high-risk HPV DNA in their oral rinse. These cases, together with 10 random samples negative for high-risk HPV DNA, were tested with the Aptima assay. A valid result was obtained for 41 of the 43 specimens (95.3%). Among the 31 cases that were positive for high-risk HPV DNA and had a valid Aptima result, 4 (12.9%) were positive for HPV mRNA. HPV mRNA was not detected in any of the samples negative for high-risk HPV DNA. HPV mRNA is detectable in oral rinses of cancer-free subjects. Oral HPV mRNA testing may be useful in the screening and/or early detection of HPV-driven OPC by possibly identifying active and transforming oral infections. The testing of individuals at increased risk for HPV-related OPC via simply and noninvasively collected oral specimens is an attractive option for future screening strategies.

328P - Unlocking the potential of saliva-based test to detect HPV-driven oropharyngeal cancer

Conference abstract published in Abstract Book of ESMO Asia Congress 23–25 November 2018, Singapore

Smoking impact on HPV driven head and neck cancer’s oncological outcomes?

IntroductionHPV-driven oropharyngeal cancer (OPC) patients have a better prognosis than their HPV-negative counterparts but several studies have suggested that among HPV-positive patients those with a smoking history had worse oncological outcomes. The aim of our study is to characterize the interplay between tobacco consumption, patient and disease characteristics, and disease control. Materials and methodsAll patients diagnosed with HPV-driven OPC and treated with curative intent between 2007 and 2009 and 2011–2016 at Gustave Roussy cancer center were included (n = 282). Demographic, clinical, morphological and tobacco consumption were correlated with oncologic outcomes. Results157 (56%) patients had a positive smoking history, including 23.8% who were smoking at the time of diagnosis and 37.6% who had a tobacco consumption exceeding 20 pack-years. In multivariate analysis, the strongest prognostic factor for survival was smoking status at cancer diagnosis, with a hazard ratio (HR) for non-smokers compared to smokers of 0.25 ([0.12, 0.50], p = 0.0001). Smoking history, either more than 20 pack-years or smoking at diagnosis, was associated with local relapse and distant relapse. There was no difference in terms of comorbidity (p = 0.32) and radiotherapy duration (p = 0.93) according to tobacco consumption. DiscussionSmoking is frequent among patients with HPV-driven OPC and increases the risk of death and oncologic failure.

Does smoking alter the mutation profile of human papillomavirus–driven head and neck cancers?

BackgroundHuman papillomavirus (HPV)-driven oropharyngeal cancer (OPC) patients are characterised by a better prognosis than their HPV-negative counterparts. However, this significant survival advantage is not homogeneous and among HPV-positive patients those with a smoking history have a significantly increased risk of oncologic failure. The reason why tobacco consumption impacts negatively the prognosis is still elusive. Tobacco might induce additional genetic alterations leading to a more aggressive phenotype. The purpose of this study was to characterise the mutational profile of HPV-positive OPCs by smoking status. We hypothesise a higher frequency of mutations affecting smokers. MethodsTargeted next-generation sequencing of 39 genes that are recurrently mutated in head and neck cancers (HNCs) caused by tobacco/alcohol consumption was performed in 62 HPV-driven OPC cases including smokers and non-smokers. ResultsThe study population included 37 (60%) non-smokers and 25 (40%) smokers. Twenty (32%) patients had no mutation, 14 (23%) had 1 mutation and 28 (45%) had 2 or more mutations. The most commonly mutated genes regardless of tobacco consumption were PIK3CA (19%), MLL2 (19%), TP53 (8%), FAT 1 (15%), FBXW7 (16%), NOTCH1 (10%) and FGFR3 (10%). Mutation rate was not significantly different in smokers compared with non-smokers even when analyses focused on heavy smokers (>20 pack-years vs. <20 pack-years). Similarly, there was no significant difference in mutations patterns according to tobacco consumption. ConclusionIn HPV-positive patients, smoking does not increase the mutation rate of genes that are recurrently mutated in traditional HNC. Additional studies are warranted to further describe the molecular landscape of HPV-driven OPC according to tobacco consumption.

Mutational profile of HPV-driven head and neck cancers according to tobacco consumption.

e17535 Background: HPV-driven oropharyngeal cancer (OPC) patients are characterized by a better prognosis than their HPV-negative counterparts. However, this significant survival advantage is not homogeneous and studies have highlighted that among HPV-positive patients those with a smoking history have a significantly increased risk of disease progression and death compared to those who have never smoked. The reason why tobacco consumption impacts negatively the prognosis is still elusive. Tobacco might induce additional genetic alterations leading to a more aggressive phenotype. The purpose of this study is to characterize the mutational profile of HPV-positive OPC by smoking status. We hypothesize a higher frequency of mutations affecting among smokers. Methods: Targeted next-generation sequencing of 38 oncogenes/tumor suppressor genes that are commonly mutated in cancers caused by tobacco/alcohol consumption was performed in 62 HPV-driven OPC cases stratified by smoking status. Results: The study population included 37 (60%) non-smokers and 25 (40%) smokers distributed as follows: 1 (4%) patient smoked &lt;10 pack-year (PY), 8 (32%) patients between 10-20 PY and 16 (64%) &gt;20 PY. Twenty (31%) patients had no mutation, 14 (23%) had 1 mutation and 28 (46%) had 2 or more mutations. The most commonly mutated genes regardless of tobacco consumption were PIK3CA (20%), MLL2 (20%), TP53 (8%), FAT 1 (15%), FBXW7 (16%), NOTCH 1 (9%) and FGFR3 (9%). Mutation rate was not significantly different in smokers compared to non-smokers even when analyses focused on heavy smokers (&gt;20 pack-years compared to &lt;20 pack-years). Similarly there was no significant difference in mutations patterns according to tobacco consumption. The 3 years overall survival, disease-specific-survival and loco-regional-control rates for the whole cohort are respectively 88% (95% CI: 76.4-94.1), 88% (95% CI: 76.4-94.1) and 80.6% (95% CI: 67.5-88.8). Despite a median follow-up was 4.5 years (6 months to 11.7 years), the few number of events (13 relapses, 13 deaths including 10 due to OPC) precludes detailed prognosis analyses. Conclusions: HPV-driven OPC patients with a smoking history have a comparable mutational rate than non-smokers. Smoking impact on the prognosis isn’t attributable to the mutational burden. Further studies are warranted.

Kinetics of the Human Papillomavirus Type 16 E6 Antibody Response Prior to Oropharyngeal Cancer.

In a European cohort, it was previously reported that 35% of oropharyngeal cancer (OPC) patients were human papillomavirus type-16 (HPV16) seropositive up to 10 years before diagnosis vs 0.6% of cancer-free controls. Here, we describe the kinetics of HPV16-E6 antibodies prior to OPC diagnosis. We used annual serial prediagnostic blood samples from the PLCO Cancer Screening Trial. Antibodies to HPV were initially assessed in prediagnostic blood drawn at study enrollment from 198 incident head and neck cancer patients (median years to cancer diagnosis = 6.6) and 924 matched control subjects using multiplex serology, and subsequently in serial samples (median = 5/individual). Available tumor samples were identified and tested for HPV16 RNA to define HPV-driven OPC. HPV16-E6 antibodies were present at baseline in 42.3% of 52 OPC patients and 0.5% of 924 control subjects. HPV16-E6 antibody levels were highly elevated and stable across serial blood samples for 21 OPC patients who were seropositive at baseline, as well as for one OPC patient who seroconverted closer to diagnosis. All five subjects with HPV16-driven OPC tumors were HPV16-E6-seropositive, and the four subjects with HPV16-negative OPC tumors were seronegative. The estimated 10-year cumulative risk of OPC was 6.2% (95% confidence interval [CI] = 1.8% to 21.5%) for HPV16-E6-seropositive men, 1.3% (95% CI = 0.1% to 15.3%) for HPV16-E6-seropositive women, and 0.04% (95% CI = 0.03% to 0.06%) among HPV16-E6-seronegative individuals. Forty-two percent of subjects diagnosed with OPC between 1994 and 2009 in a US cohort were HPV16-E6 seropositive, with stable antibody levels during annual follow-up for up to 13 years prior to diagnosis. Tumor analysis indicated that the sensitivity and specificity of HPV16-E6 antibodies were exceptionally high in predicting HPV-driven OPC.

A Survey of Wisconsin Pediatricians' Knowledge and Practices Regarding the Human Papillomavirus Vaccine.

Objective The human papillomavirus (HPV) is common and carries a significant burden of disease. This is increasingly apparent in males with the rising incidence of HPV-related oropharyngeal cancer. Unfortunately, vaccination rates remain poor and are lowest in males. It is unclear if pediatricians are aware of the alarming rise of HPV-mediated head and neck cancers and the disproportionate effect on males. Study Design This investigation used a cross-sectional descriptive survey research design. Setting The survey was developed by investigators in the University of Wisconsin Division of Otolaryngology. Subjects and Methods The survey was distributed to 831 members of the Wisconsin Chapter of the American Academy of Pediatrics. Results A total response rate of 49.6% was achieved. Most supported routine vaccination in both sexes. Females are regarded as being at higher risk of an HPV-related cancer and are more often recommended vaccination. Most providers are unaware of the magnitude of HPV-related oropharyngeal cancer and the greater affliction in males. Conclusions Male vaccination is overwhelmingly supported by Wisconsin pediatricians, yet there is a preponderance toward vaccinating females, who are perceived as having greater risk for HPV-associated disease. This is likely because providers are unaware of the magnitude of HPV-driven oropharyngeal cancer and its predilection for males. A lack of provider awareness, in combination with out-of-date education material for parents, likely contributes to poor vaccination rates in males.

Human papillomavirus (HPV) 16 antibodies at diagnosis of HPV-related oropharyngeal cancer and antibody trajectories after treatment

ObjectivesDespite the fact that HPV-driven oropharyngeal cancer (HPV-OPC) has relatively low recurrence rates, intensive post-therapy monitoring remains the standard of care. Post-treatment biomarkers are needed to risk stratify HPV-OPC patients for more individualized surveillance intensity and which remain at higher recurrence risk. Materials and Methods115 HPV-OPC patients (ascertained by p16 immunohistochemistry and/or in-situ hybridization) from a multicenter prospective case study (HOTSPOT) had blood collected at diagnosis, and 64 of these also had blood collected at post-treatment follow-up visits for up to two years. Samples were centrally tested for antibodies to the L1, E1, E2, E4, E6, and E7 proteins of HPV16. ResultsAt diagnosis, most HPV-OPC cases were seropositive to HPV16 E6 (85%). In post therapeutic samples, HPV16 antibody level decreased slowly over time, but only 3 (of 51 cases seropositive at enrollment) dropped low enough to be classified as seronegative. At 3years after diagnosis, cumulative risk of recurrence was 10.2% and 0% in HPV16 E6 seropositive and E6 seronegative HPV-OPC cases, respectively (p=0.18). Risk of recurrence was increased, although not statistically significant, in those with higher HPV16 E6 antibody levels at diagnosis (per log antibody level, hazard ratio [HR]=1.81, 95%CI=0.47–6.92). ConclusionThis study confirms the high seroprevalence of HPV oncogenic antibodies at diagnosis of HPV-OPC. HPV16 E6 antibody levels decrease after treatment, but most cases remain seropositive for up to two years. HPV16 E6 antibody levels at diagnosis did not appear to be a strong predictor of recurrence.

Diagnosis of HPV driven oropharyngeal cancers: Comparing p16 based algorithms with the RNAscope HPV-test

BackgroundAccurate identification of HPV-driven oropharyngeal cancer (OPC) is a major issue and none of the current diagnostic approaches is ideal. An in situ hybridization (ISH) assay that detects high-risk HPV E6/E7 mRNA, called the RNAscope HPV-test, has been recently developed. Studies have suggested that this assay may become a standard to define HPV-status. MethodsTo further assess this test, we compared its performance against the strategies that are used in routine clinical practice: p16 immunohistochemistry (IHC) as a single test and algorithms combining p16-IHC with HPV-DNA identification by PCR (algorithm-1) or ISH (algorithm-2). Results105 OPC specimens were analyzed. The prevalence of HPV-positive samples varied considerably: 67% for p16-IHC, 54% for algorithm-1, 61% for algorithm-2 and 59% for the RNAscope HPV-test. Discrepancies between the RNAscope HPV-test and p16-IHC, algorithm-1 and 2 were noted in respectively 13.3%, 13.1%, and 8.6%.The 4 diagnostic strategies were able to identify 2 groups with different prognosis according to HPV-status, as expected. However, the greater survival differential was observed with the RNAscope HPV-test [HR: 0.19, 95% confidence interval (CI), 0.07–0.51, p=0.001] closely followed by algorithm-1 (HR: 0.23, 95% CI, 0.08–0.66, p=0.006) and algorithm-2 (HR: 0.26, 95% CI, 0.1–0.65, p=0.004). In contrast, a weaker association was found when p16-IHC was used as a single test (HR: 0.33, 95% CI, 0.13–0.81, p=0.02). ConclusionsOur findings suggest that the RNAscope HPV-test and p16-based algorithms perform better that p16 alone to identify OPC that are truly driven by HPV-infection. The RNAscope HPV-test has the advantage of being a single test.

Low prevalence of HPV-driven head and neck squamous cell carcinoma in North-East Italy

ObjectivesTo investigate the frequency of Human papillomavirus (HPV)-driven head and neck squamous cell carcinoma (HNSCC) among patients living in North-East Italy, by assessing HPV-DNA positivity in all tumors and additional markers whenever possible. Material and methodsHPV types, viral load, viral RNA, HPV16/18 E6 protein and p16INK4a and pRb expression were determined in primary tumor tissues from 247 HNSCC patients. Tumor-specific HPV seropositivity was analyzed in 102 patients. ResultsTumor HPV-DNA prevalence was 8.5% overall (21/247) and 27% in oropharynx (17/63). HPV16 accounted for 95% of all HPV types found. Among HPV-DNA+ tumors, type-concordant HPV E6*I RNA prevalence was 79%. HPV DNA+ RNA+ tumors showed high viral load, up-regulated p16INK4a, down-regulated pRb and presence of HPV16 E6 protein. Eight cases showed tumor-specific HPV seropositivity, all type-concordant with the tumor. Tumors were defined as HPV-driven when positive for HPV-DNA plus 2 additional HPV transformation-related markers. ConclusionRelative prevalence of HPV-driven tumors (14 HPV16, 1 HPV58) was 6% overall and 20% among oropharyngeal cancers. In the oropharynx the HPV-driven group showed a trend for better survival versus the HPV-negative group.The relative prevalence of HPV-driven oropharyngeal cancer is low in North-East Italy as compared to Western and Northern Europe.

Diagnosis of HPV driven head and neck cancer: Comparing p16 based algorithms with the RNAscope HPV-test.

6040Background: Accurate identification of HPV-driven oropharyngeal cancer (OPC) is a major issue and none of the current diagnostic approaches is ideal. An in situ hybridization (ISH) assay that d...

Characterization of human papillomavirus antibodies in individuals with head and neck cancer

BackgroundHuman papillomavirus type 16 (HPV16) E6 antibodies are a promising biomarker of oropharyngeal cancer (OPC); however, seropositivity among non-OPC cases is not well characterized. MethodsPre-treatment sera from 260 (38 OPC, 222 non-OPC) incident head and neck cancers diagnosed at the University of Pittsburgh between 2003 and 2006 were tested for HPV16 (L1,E1,E2,E4,E6,E7) and non-HPV16 E6 (HPV6,11,18,33) antibodies. Sensitivity and specificity of HPV16 E6 antibodies for HPV-driven tumors was evaluated among tumors with known HPV status (n=25). Results63.2% of OPC versus 27.5% of non-OPC cases were HPV16 seropositive; HPV16 E6 seroprevalence was 60.5% and 6.3% respectively, odds ratio 22.8 (95% confidence interval [CI] 9.8⿿53.1). Sensitivity and specificity of HPV16 E6 antibodies for HPV-driven OPC was 100% [95% CI: 50⿿100%; n=6] and 100% [95% CI: 60⿿100%, n=4] compared to 0% (n=2) and 0% (n=13) for non-OPC cases. ConclusionsHPV16 antibodies were significantly more common in OPC versus non-OPC cases, particularly HPV16 E6 antibodies.

HPV vaccination and the prevention of oropharyngeal cancer: two steps forward and one to the side?

In September 2014 the Joint Committee on Vaccination and Immunisation (JCVI) recommended a small extension to the provision of human papillomavirus (HPV) vaccination beyond girls aged 12–13 years. This applied to men who have sex with men and shows a limited receptiveness to existing evidence in relation to the concept of ‘herd protection’ as well as the increasing incidence of HPV-driven oropharyngeal cancer (in both sexes, but men more than women) and anal cancer (in both sexes but particularly in women).