Characterization of High-risk Oncogenic Human Papillomavirus Genotypes in Histologically Confirmed Ear, Nose and Throat (Ent) Cancers in Burkina Faso.

BACKGROUND: High-risk human papillomavirus (HPV-HR) infections are responsible for 99.99% of cervico-uterine cancers and 50% of carcinomas of the oropharynx. OBJECTIVE: To characterize high-risk HPV genotypes (HPV-HR) in histologically confirmed ear, nose and throat (ENT) cancers in Ouagadougou. METHODS: One hundred and twenty-eight archived tissues from the ENT sphere, obtained over the last ten years (2007 to 2017) and histologically diagnosed in anatomy and pathology-cytology laboratories in Ouagadougou were included. These tissues were dewaxed with xylene; HPV DNA extraction was performed and HPV-HR were researched by real-time multiplex PCR. RESULTS: Among the fourteen HPV-HR genotypes tested for, seven were identified. The prevalence of HPV-HR infection was 15.6%. The most common genotypes were: HPV56 (45%) and HPV33 (20%). Squamous cell carcinomas accounted for 75% of cases, followed by lymphomas for 10%. The age range was between 5 and 80 years. CONCLUSION: The results show the involvement of a diversity of HPV-HR genotypes and a high frequency of HPV56 and HPV33 in ENT cancers in Ouagadougou, Burkina Faso. The appropriate HPV vaccination will considerably reduce the number of these cancers.
.

[The detection of the human papilloma virus during hyperplastic processes in the nose, ears and throat]. / Obnaruzhenie virusa papillomy cheloveka pri giperplasticheskikh protsessakh v LOR-organakh.

The objective of the present work was to carry out the virological and histological studies of various neoplastic and hyperplastic processes in the nose, ears, and throat with a view to identifying the presence of human papilloma virus and Epstein-Barr virus. The brush biopsies and remote neoplasms obtained from 18 patients (including 2 children and 16 adults) presenting with various ENT diseases and tumours were available for the virological investigation with the use of the polymerase chain reaction (PCR) and a system MY09-MY11 degenerate primers . The histological study of biopsies and remote neoplasms was carried out by means of conventional light microscopy. The virological and histological studies conducted in parallel confirmed the diagnostic significance of morphological changes at the tissue and cellular levels caused by the human papilloma virus.

Characteristics of Human Papillomavirus-Associated Head and Neck Cancers in a Veteran Population.

IMPORTANCE: The US veteran population represents a unique cohort of patients in whom human papillomavirus (HPV)-related head and neck squamous cell carcinoma (HNSCC) has yet to be investigated. OBJECTIVE: To investigate the incidence and characteristics of HPV-positive HNSCC within the veteran population. DESIGN, SETTING, AND PARTICIPANTS: Retrospective medical record review including patients with HNSCC diagnosed between January 1, 2010, and November 15, 2013, from the VA Greater Los Angeles Healthcare System. Data were collected between November 16, 2013, and June 19, 2014, and analyzed between June 20, 2014, and March 26, 2015. EXPOSURES: Chemoradiation therapy, radiation therapy, surgery, or no treatment. MAIN OUTCOMES AND MEASURES: We determined HPV positivity by p16 testing. Demographic and clinicopathologic information and overall survival were extracted from medical records. RESULTS: We identified 150 patients with the diagnosis of HNSCC. Sixty-nine patients had HPV-positive tumors (46%), and 65 (43%) had HPV-negative tumors (16 did not have HPV testing). Age at diagnosis ranged from 44 to 94 years (mean, 64.6 [SD, 8.0] years), and median (interquartile range) follow-up was 16.7 (8.7-27.3) years. Tumor location differed significantly between the 2 groups, with an HPV-positive predominance in the oropharynx (43 of 57 [75%]; P < .001). The HPV-positive patients were more likely to be treated primarily with combined chemoradiation therapy than radiation therapy or surgery (P < .001). T4 tumors had a nearly 9 times greater rate of mortality compared with T1 tumors (HR, 8.52 [95% CI, 2.60-18.40; P < .001); N3 disease was associated with 7.18 times greater mortality (HR, 7.18 [95% CI, 1.99-12.26]; P < .001) compared with N1 disease; and M1 disease was associated with 6.0 times greater mortality (HR, 5.99 [95% CI, 2.59-13.81]; P < .001). There were 42 total deaths during follow-up, 25 in the HPV-negative group and 17 in the HPV-positive group, with a nonsignificantly higher overall survival among HPV-positive patients independent of alcohol or tobacco use history (P = .09). CONCLUSIONS AND RELEVANCE: Previous studies have found that the proportion of HPV-positive HNSCC in the general population ranges between 20% and 75%. Although the incidence of HPV-positive HNSCC in the Veterans Affairs population is comparable, these patients have unique risk factors and demographic characteristics that may suggest different prognostic factors for HPV-positive HNSCC in this population. Nonetheless, HPV-positive tumors still seem to portend a better overall prognosis regardless of alcohol or tobacco history among the Veterans Affairs population.

Oncolytic activity of reovirus in HPV positive and negative head and neck squamous cell carcinoma.

BACKGROUND: The management of patients with advanced stages of head and neck cancer requires a multidisciplinary and multimodality treatment approach which includes a combination of surgery, radiation, and chemotherapy. These toxic treatment protocols have significantly improved survival outcomes in a distinct population of human papillomavirus (HPV) associated oropharyngeal cancer. HPV negative head and neck squamous cell carcinoma (HNSCC) remains a challenge to treat because there is only a modest improvement in survival with the present treatment regimens, requiring innovative and new treatment approaches. Oncolytic viruses used as low toxicity adjunct cancer therapies are novel, potentially effective treatments for HNSCC. One such oncolytic virus is Respiratory Orphan Enteric virus or reovirus. Susceptibility of HNSCC cells towards reovirus infection and reovirus-induced cell death has been previously demonstrated but has not been compared in HPV positive and negative HNSCC cell lines. OBJECTIVES: To compare the infectivity and oncolytic activity of reovirus in HPV positive and negative HNSCC cell lines. METHODS: Seven HNSCC cell lines were infected with serial dilutions of reovirus. Two cell lines (UM-SCC-47 and UM-SCC-104) were positive for type 16 HPV. Infectivity was measured using a cell-based ELISA assay 18 h after infection. Oncolytic activity was determined using an alamar blue viability assay 96 h after infection. Non-linear regression models were used to calculate the amounts of virus required to infect and to cause cell death in 50% of a given cell line (EC50). EC50 values were compared. RESULTS: HPV negative cells were more susceptible to viral infection and oncolysis compared to HPV positive cell lines. EC50 for infectivity at 18 h ranged from multiplicity of infection (MOI) values (PFU/cell) of 18.6 (SCC-9) to 3133 (UM-SCC 104). EC50 for cell death at 96 h ranged from a MOI (PFU/cell) of 1.02×10(2) (UM-SCC-14A) to 3.19×10(8) (UM-SCC-47). There was a 3×10(6) fold difference between the least susceptible cell line (UM-SCC-47) and the most susceptible line (UM-SCC 14A) EC50 for cell death at 96 h. CONCLUSIONS: HPV negative HNSCC cell lines appear to demonstrate greater reovirus infectivity and virus-mediated oncolysis compared to HPV positive HNSCC. Reovirus shows promise as a novel therapy in HNSCC, and may be of particular benefit in HPV negative patients.

Hybrid Capture 2 is as effective as PCR testing for high-risk human papillomavirus in head and neck cancers.

High-risk human papillomavirus (HPV) infection is a common cause of oropharyngeal squamous cell carcinoma, especially in young male nonsmokers. Accurately diagnosing HPV-associated oral cancers is important, because they have a better prognosis and may be treated differently than smoking-related oral carcinomas. Various methods have been validated to test for high-risk HPV in cervical tissue samples, and they are in routine clinical use to detect dysplasia before it progresses to invasive disease. Similarly, future screening for HPV-mediated oropharyngeal dysplasia may identify patients before it progresses. Our objective was to compare 4 of these methods in a retrospective series of 87 oral and oropharyngeal squamous cell carcinomas that had archived fresh-frozen and paraffin-embedded tissue for evaluation. Patient age, sex, smoking history, and tumor location were also recorded. DNA prepared from fresh-frozen tissue was tested for HPV genotypes by multiplex polymerase chain reaction analysis, and high-risk HPV screening was carried out using Hybrid Capture 2 and Cervista. Histologic sections were immunostained for p16. HPV-positive outcome was defined as agreement between at least 2 of the 3 genetic tests and used for χ analysis and calculations of diagnostic predictive value. As expected, high-risk HPV-positive oral cancers were most common in the tonsil and base of the tongue (oropharynx) of younger male (55 vs. 65 y) (P=0.0002) nonsmokers (P=0.01). Most positive cases were HPV16 (33/36, 92%). Hybrid Capture 2 and Cervista were as sensitive as polymerase chain reaction and had fewer false positives than p16 immunohistochemical staining.

[Advances in head and neck cancers on behalf of the French Intergroup ORL and GORTEC]. / Évolution des concepts dans les cancers des voies aérodigestives supérieures, sous l'égide de l'Intergroupe ORL (GORTEC, GETTEC, GERCOR).

Head and neck cancers are the fifth among the most common cancers in France. Two thirds of cases occur at an advanced stage. For advanced disease, progression-free survival, despite undeniable progress, remains below 50% at three years. The last 20 years have been marked by the necessity to identify situations where less intense surgery and/or radiotherapy and/or chemotherapy is possible without jeopardizing the prognosis, and situations where a therapeutic intensification is necessary and results in a gain in survival while better preserving function with less toxicity. French cooperative groups gathering radiation oncologists (GORTEC), surgeons (GETTEC) and medical oncologists or physicians involved in the management of systemic treatments in head and neck cancers (GERCOR) are now belonging to the INCa-labelled Intergroup ORL to deal with the challenges of head and neck cancers.

[HPV-associated head and neck cancer. The basics of molecular and translational research]. / Humane Papillomaviren bei Kopf-Hals-Karzinomen. Molekulare und translationale Grundlagen.

Translational research refers to the interfaces between preclinical research and targeted short- and medium-term developments through to clinical standards. There are two distinct groups of oropharyngeal malignancies: those caused by tobacco and alcohol abuse and those caused by HPV infection. Although the prognosis of patients in the latter group is significantly better, this is not taken into consideration in the choice of treatment. However, less intensive use of radiotherapy, chemotherapy, or surgery, as well as targeted multimodal therapeutic approaches, is under research. This article summarizes the main events in the HPV life cycle, with emphasis on carcinogenic mechanisms and potential new molecular targets. Identifying distinct tumor entities of the oropharynx enables the design and development of new preventive and therapeutic strategies to reduce the incidence and mortality of HPV-associated oropharyngeal cancers in the near future.

Human papillomavirus in metastatic lymph nodes from unknown primary head and neck squamous cell carcinoma.

OBJECTIVE: Determine human papillomavirus (HPV) incidence in unknown primary squamous cell carcinomas (SCCa) of the head and neck and assess if HPV status influenced survival. STUDY DESIGN: Historical cohort study. SETTING: Tertiary care center. SUBJECTS: Patients with unknown primary SCCa despite a complete workup who underwent neck dissection or excisional biopsy and postoperative comprehensive ± chemoradiotherapy between 2002 and 2009. METHODS: HPV fluorescence in situ hybridization (FISH) and p16(INK4a) immunohistochemistry (p16 IHC) were performed. Results were compared with survival, age, race, gender, tobacco use, alcohol use, and nodal stage. RESULTS: Twenty-five patients met the inclusion criteria, of whom 88% were >10 pack year tobacco users. Twenty-eight percent were HPV-positive defined by both p16+ and FISH+. Five-year overall survival was 66.7% in HPV-positive and 48.5% in HPV-negative patients (P = .35). Similarly, 5-year disease-free survival rates were 66.7% in HPV-positive and 48.5% in HPV-negative patients (P = .54). All 3 HPV-positive nonsmokers were survivors, but this was not significant because of the small sample size (P > .05). No other characteristics were associated with survival (P > .05). CONCLUSION: Twenty-eight percent of metastatic lymph nodes from occult primary tumors were HPV positive. There was no survival difference associated with HPV status. Most of the HPV-positive patients in this study were tobacco users who had similar survival to HPV-negative patients, so caution should be used in interpreting HPV status in these patients.

Histopathologic findings of HPV and p16 positive HNSCC.

OBJECTIVE: Human papilloma virus (HPV) and p16INKa (p16) positivity in head and neck squamous cell carcinomas (HNSCCs) is currently thought to be an encouraging prognostic indicator. However, the histopathologic changes responsible for this behavior are poorly understood. It is our objective to elucidate these histopathologic characteristics to help define the clinical utility of these markers. DESIGN: Retrospective cohort study. METHODS: 71 HNSCC tumors between July 1, 2008 and August 30, 2009 were examined for HPV, p16, and epidermal growth factor receptor (EGFR). Specified pathologic features were examined: perivascular invasion (PVI), perineural invasion (PNI), grade of squamous differentiation, basaloid classification. RESULTS: HPV and p16 had no direct impact on perineural or perivascular invasion. However, HPV and p16 were strongly predictive of poorly differentiated tumors, as well as basaloid squamous cell carcinoma (SCCA) (P < .001). Additionally, upon multivariate analysis, HPV(+) and p16(+) tumors had an increased risk of nodal metastasis (HPV: odds ratio [OR] = 23.9 (2.2, 265.1) p = .01; p16: OR = 6.5 (1.4, 31.2) p = .02; PVI: OR = 6.0 (1.6, 22.8) p < .01). The area under the curve (AUC) of receiver operating characteristic (ROC) curves demonstrated improved predictive value for lymph node metastasis above standard H&E histopathologic features (76.7%) for both HPV (83.2%) and p16 (81.3%) individually. CONCLUSIONS: HPV(+) and p16(+) are highly predictive for poorly differentiated tumors and basaloid SCCA. Additionally, HPV and p16 positivity demonstrate superior predictive value for lymph node metastasis above standard H&E histopathologic features. Although exact recommendations should be tempered by considerations of primary tumor subsite, T-stage, and depth of invasion, head and neck multidisciplinary teams should strongly consider aggressive lymph node treatment for any HPV(+) or p16(+) tumor.

Human papillomavirus status in head and neck cancer: the ethics of disclosure.

Human papillomavirus (HPV) is an emerging causative factor for squamous carcinoma of the oropharynx and perhaps other head and neck cancers. There is a great deal of uncertainty regarding the clinical significance and implications of HPV status in this patient population. As a result, there is no established protocol for informing patients of the potential link between viral infection and their cancer. This paper discusses some of the ethical issues involved with informing head and neck cancer patients of their HPV status, recognizing the dilemma posed by unresolved clinical questions and the need to respect the autonomy of patients by disclosing relevant information.

[Prognosis and predictive factors in head-and-neck cancers]. / Facteurs pronostiques et prédictifs des cancers des voies aérodigestives supérieures.

The head-and-neck squamous-cell carcinomas (HNSCC) represent in order of frequency the fourth leading cause of cancer deaths among men in France. The term HNSCC includes various anatomopathological and clinical entities with different evolution patterns. For this reason, it is necessary to identify prognostic and predictive factors able to help in the choice of the treatment. The clinical factors with a prognostic value are the tumor location, the tumor size and the lymph node status. The degree of differentiation is the most important histologic factor. More recently, the identification of molecular factors has opened the way to new therapies. Thus, the overexpression of EGFR is associated with a poor prognosis. Its inhibition improves the survival of patients. p53 mutations and cyclin D1 amplification are actually subject to intensive research. The tumors associated with HPV infection are distinguishable by a better prognosis. 18-alpha-FDG positron emission tomography emerges as a useful tool for the therapeutic evaluation. The evolution of surgical techniques, the development of induction chemotherapy regimen or concurrent ones with radiotherapy and new techniques of conformal irradiation also results in a better locoregional control.

[HPV in non-gynecological tumors]. / HPV bei nichtgynäkologischen Tumoren.

Human papilloma viruses (HPVs) are the main tumor viruses in humans and have been identified in gynecological malignancies, especially carcinomas of the uterine cervix and their precursor lesions. In addition, they are frequently observed in other anogenital tumors such as vulva/vagina, anal and penis carcinoma. Furthermore, the potential association with head and neck cancer, in particular tonsillar carcinoma, is now well documented. However, there are controversial reports on the detection of HPV in various other tumors; these are summarized in the present report. Data revealed that apart from the heart and the kidney, the virus has been found in all other organs that have been analyzed so far, i.e., prostate, urinary bladder, oral cavity, larynx, esophagus, stomach, colon, liver, vagina/vulva, endometrium, ovary, breast, penis, anus, skin, and lung. Some of the detection rates are remarkable, e.g., colon cancer up to 97%, lung cancer 80%, and breast cancer 74%. They point to geographical differences in the incidence of HPV in different populations, but also highlight the need for validation of the results. HPV is nevertheless an important biomarker in molecular tumor diagnostics. Firstly, it is useful for the differentiation of a secondary squamous cell carcinoma from a metastasis. Secondly, HPV-positive carcinomas not only have a distinct etiology but also a particular morphological phenotype. Overall, they are characterized by different tumor biology, such as, for example, increased sensitivity to radiotherapy.

Effects of cidofovir on a novel cell-based test system for recurrent respiratory papillomatosis.

BACKGROUND: Cidofovir has been reported to have activity against human papillomavirus (HPV) type 16, but no laboratory studies have been performed on HPV type 6, the main cause of recurrent respiratory papillomatosis (RRP). METHODS: HPV6b E6 cDNA-based C33A (non-HPV cervical carcinoma) cell line was produced. Two different doses of cidofovir were applied to parent C33A, C33AT6E6, and C33AT16E6 (HPV 16). Growth and flow cytometry analysis were performed. RESULTS: Polymerase chain reaction confirmed HPV6 E6 expression in C33AT6E6 cells. High-dose cidofovir was found to be toxic to all cell lines. Low-dose exposure was found to be toxic to C33AT16E6 cells at 3 days, whereas C33A and C33AT6E6 showed minimal toxicity at 6 days and earlier recovery following drug withdrawal. CONCLUSIONS: Cidofovir showed nonspecific toxicity against all 3 cell lines tested. HPV16 E6 expressing cells were more sensitive than parent or HPV6 E6 expressing cells. Cidofovir has no selective advantage for the RRP-related HPV6 E6 expressing cell line.

[The detection of human papilloma virus in juvenile-onset respiratory papillomatosis].

OBJECTIVE: To investigate the relationship between juvenile-onset respiratory papillomatosis (JOP) and human papilloma virus (HPV6, HPV11), and the immune function of patients. METHOD: Fluorescence quantitative PCR (FQ-PCR) which combines PCR and fluorescence probe hybridization was used to detect HPV6 , HPV11 DNA in 130 cases. Of these, 68 cases were used Flow Cytometry to measure CD3, CD4, CD8. RESULT: One hundred and fifteen of 130 cases were HPV6, HPV11 DNA positive, the average copy was 5.68 +/- 2.65. For 68 cases,the average percent of CD3,CD4,CD8 were 62.73 +/- 8.63, 30.54 +/- 7.05, 26.08 +/- 6.93, respectively. To compare with control group, there was not statistical significance. CONCLUSION: FQ-PCR is a convenient, accurate and specific method which can quantify the amount of pathogenic germs from 10(1) to 10(10), and is also a reliable factor to predicate clinical diagnose and cure.

Human papillomaviruses in lymph node neck metastases of head and neck cancers.

CONCLUSION: The results of this study corroborate earlier findings that human papillomavirus (HPV)16 is the most prevalent type of HPV in squamous cell carcinomas of the head and neck (SCCHNs) and reinforce a possible influence of HPV on SCCHN progression by showing that the majority of HPV-positive patients harbor HPV16 (or HPV33) both in their primary tumors and in lymph node neck metastases (LNNMs). OBJECTIVE: HPVs are causally associated with carcinomas of the uterine cervix and have also been linked to a subset of SCCHNs. In order to further investigate the predicted causative role of HPV in SCCHNs, we analyzed pairs of primary tumors and LNNMs or LNNMs alone for the presence of HPV DNA using polymerase chain reaction (PCR). MATERIAL AND METHODS: DNA was extracted from fresh frozen tissue samples of primary tumors and the corresponding LNNMs of 18 patients and from LNNMs alone in 17 patients. For the detection and typing of HPV, PCR was performed using both type-specific and consensus primer pairs, followed by Southern hybridization and, in selected cases, sequencing of the PCR products. RESULTS: Of the 35 patients investigated, 22 (63%) were found to have HPV DNA in their tumors: HPV16 DNA in 21 cases and HPV33 in 1. The highest HPV prevalence was detected in tumors of Waldeyer's tonsillar ring (8/9 patients; 89%). Of the 18 patients in whom primary tumors and LNNMs were analyzed, 7 (39%) were HPV-positive in both samples (HPV16, n = 6; HPV33, n = 1), in 3 (17%) the primary tumors were HPV-negative and the LNNMs HPV16-positive and in 1 (5.5%) the primary tumor contained HPV16 and the LNNM was negative. Interestingly, of the 7 patients in whom LNNMs had been detected only several months after diagnosis and treatment of the primary tumors, only 1 showed infection with HPV (HPV33).

Role of human papillomavirus in the development of head and neck squamous cell carcinomas.

OBJECTIVE: To review the literature on the role of oncogenic human papillomaviruses (HPVs) in the carcinogenesis of the head and neck mucosa. MATERIAL AND METHODS: Molecular and epidemiological studies concerning the high-risk HPV types and their role in carcinogenesis in the head and neck region were screened. RESULTS: Different studies revealed that: (i) 15-25% of head and neck squamous cell carcinomas (HNSCCs) are clonally associated with high risk HPV types (type 16); (ii) the oropharynx and particularly the tonsils are the most susceptible sites; (iii) patients with HPV-positive tumours present with more advanced stages of disease, are relatively younger, do not have extravagant tobacco and alcohol intake and seem to have a better survival; (iv) HPV-positive tumours are characterized by poor differentiation grade and a basaloid appearance; and (v) HPV-positive tumours exhibit integrated HPV DNA, wild-type p53, pRb downregulation and overexpression of p16INK4A. CONCLUSION: Taken together, these data support the view that HPV-harbouring HNSCC can be considered a discrete tumour entity with, moreover, a favourable prognosis. Screening of patients, especially those with tonsillar cancers, for the presence of HPV may help to further optimize treatment protocols and to provide more accurate prognostic information.

Detection of Epstein-Barr virus-derived latent membrane protein-1 gene in various head and neck cancers: is it specific for nasopharyngeal carcinoma?

OBJECTIVE: The object of the study was to determine the incidence of the presence of Epstein-Barr virus-derived latent membrane protein-1 (LMP-1) gene in various head and neck cancers by polymerase chain reaction method. STUDY DESIGN: Prospective study. METHODS: During a 5-year period, polymerase chain reaction was used to investigate the presence of LMP-1 gene in various head and neck cancers from different locations and histopathological types, noncancerous nasopharyngeal biopsy samples, and tonsillectomy specimens from patients with chronic hypertrophic tonsillitis. RESULTS: Of 202 patients enrolled in the study, 53 were diagnosed by pathological study with oropharyngeal carcinoma, 45 with nasopharyngeal carcinoma, 26 with oral cavity carcinoma, 26 with laryngohypopharyngeal carcinoma, 31 with nasopharyngeal lymphoid hyperplasia, and 21 with tonsils with lymphoid hyperplasia. After the application of polymerase chain reaction, the LMP-1 gene was not detected in any sample from oral cavity carcinoma, laryngohypopharyngeal carcinoma, or nasopharyngeal lymphoid hyperplasia or from tonsillectomy specimens but was detected in only one case of tonsillar carcinoma. On the contrary, the LMP-1 gene was detected in 43 (95.6%) of 45 samples from patients with nasopharyngeal carcinoma. The statistical analysis shows a significant association (P <.001) between the presence of LMP-1 gene and tumor localization in the nasopharynx. CONCLUSIONS: The study shows that the presence of LMP-1 gene detected by polymerase chain reaction in the tumor cell is only significantly associated with tumor located in the nasopharynx, implying that Epstein-Barr virus plays a trifling role in the tumorigenesis of carcinomas arising from other head and neck locations. The polymerase chain reaction method that was used is a potential tool for screening nasopharyngeal carcinoma.