Jan Klozar

Demographic and risk factors in patients with head and neck tumors

The association between human papillomavirus (HPV) infection and the development of head and neck cancer has been documented recently. In this study on 86 head and neck cancer patients and 124 controls, data regarding demographics, behavioral risk factors, and risks related to HPV exposure were collected. HPV detection was carried out using polymerase chain reaction in the tumors and in oral exfoliated cells, and HPV typing by a reverse line blot assay specific for 37 HPV types. Sera were tested by an enzyme‐linked immunosorbent assay specific for HPV proteins. Head and neck cancer cases report significantly more oral‐anal contact (P = 0.02) and tobacco and alcohol use than controls (P = 0.001; P = 0.02, respectively). High‐risk HPV DNA was detected in 43% of oral washings of cases and 4% of controls (P < 0.0001). The association between the presence of high‐risk HPV DNA in oral exfoliated cells and in tumor tissues was statistically significant (adjusted P < 0.0001). The prevalence of HPV‐specific antibodies was significantly higher in cases than in controls (adjusted P < 0.0001). These results provide epidemiological and immunological evidence for HR HPV as a strong risk factor (OR = 44.3, P < 0.0001) for head and neck cancer, even after controlling for age, tobacco and alcohol use. The detection of high‐risk HPV DNA in oral exfoliated cells and HPV‐specific antibodies in serum can be considered as clinically relevant surrogate markers for the presence of a HPV‐associated head and neck cancer, with a high sensitivity (83%) and specificity (88%). J. Med. Virol. 81:878–887, 2009.

HPV involvement in tonsillar cancer: Prognostic significance and clinically relevant markers

The association of high‐risk human papillomaviruses (HR HPVs) with tonsillar cancer (TC) has been documented. Because patients with HPV‐associated tumors show better survival rates, modification of their treatment regimen is being considered. It is therefore crucial to find markers for the identification of patients whose tumors are linked to viral infection. A cohort of 109 patients with primary TC was screened for HPV DNA presence in the tumor tissues and HPV‐specific antibodies in sera. Data regarding risk factors and clinical parameters were collected. Forty‐five specimens were analyzed for the expression of viral E6 and E2‐region mRNA, and the p16 and p53 protein expression status was assessed by immunohistochemistry. The overall prevalence of HPV DNA in TC tissues was 65.1%. Ninety‐three percent of HR HPV DNA‐positive samples expressed E6*I mRNA. E2‐region mRNA expression was detected in 36% of positive samples, which implies that the virus is integrated in 64% of HPV DNA/RNA‐positive tumors. p16 overexpression and the presence of antibodies specific to HPV16 E6/E7 oncoproteins correlated well with HPV DNA and RNA presence. The disease‐specific survival rate of patients with HPV DNA‐positive tumors was significantly higher than that of HPV DNA‐negative patients. In addition to providing further evidence of the involvement of HPV infection in the etiopathogenesis of a proportion of TC cases, our study demonstrates that p16 immunostaining and anti‐E6/E7 antibodies as surrogate markers of HPV involvement represent specific, sensitive and clinically accessible assays for the identification of TC patients who have a considerably better prognosis.

Nodal status is not a prognostic factor in patients with HPV‐positive oral/oropharyngeal tumors

The primary aim is to compare the prognostic parameters in patients with HPV‐positive and HPV‐negative tumors. The secondary aim is to compare the patterns of treatment failure between these groups.

Markers of HPV infection and survival in patients with head and neck tumors

The purpose of this study was to determine whether changes in human papillomavirus (HPV) DNA prevalence in oral rinses and/or HPV‐specific antibody levels in the sera of patients with oral/oropharyngeal cancer have prognostic significance. One hundred and forty‐two patients with oral/oropharyngeal tumors were enrolled. The presence of HPV DNA was assayed in tumor tissue and oral rinses and HPV‐specific antibodies were assessed in the sera. Oral rinses were collected before treatment and one year after the treatment. Sera were drawn before treatment, one month, and one year after the end of the treatment. Altogether, 59.2% of tumors were HPV positive. The presence of HPV DNA in the tumors correlated with HPV DNA positivity in oral rinses and with HPV‐specific antibodies in the sera. Out of 66 patients with HPV‐positive oral rinses at enrolment, 84.8% became negative at one‐year follow‐up, while most patients remained seropositive for HPV‐specific antigens. However, the mean titers of HPV16 E6 and/or E7 antibodies at follow‐up were significantly lower. Of 16 patients with recurrences at follow‐up (alive on second sampling), six were positive at enrolment for HPV16 E6 and/or E7 antibodies. In five of these, no decrease in antibody levels was observed. Titers of antibodies specific for HPV16 capsid antigens did not change during the follow‐up. Our data suggest that the detection of antibodies specific for the HPV 16 E6 and E7 oncoproteins may serve not only as a marker of HPV etiology, but also as a marker of recurrence and a prognostic indicator in patients with HPV‐positive tumors.

Laryngeal Papilloma—Precancerous Condition?

In a group of 179 patients treated for recurrent laryngeal papillomatosis 668 surgeries were performed in the years 1982-1995 in the Department of Otorhinolaryngology, Head and Neck Surgery in Prague. The group was divided into 77 patients with a juvenile form of papillomatosis and 102 patients with an adult form. The adult form was then divided into a multiple (65 patients) and solitary form (37 patients). Three patients with a juvenile form of papillomatosis were irradiated in advance. None of these patients developed a carcinoma. There were 3 cases (1.7%) of carcinoma in the whole group of patients with histologically verified papillomas during repeated previous surgeries. All 3 patients with malignancy had an adult form of papillomatosis, two with a multiple form and one with a solitary form. The intervals between the first treatment for papilloma and the diagnosis of carcinoma was 8, 3 and 2 years.The rates of malignant transformation of papillomas vary in the literature. We suppose, that because of the generally long interval between the diagnosis of papilloma and that of carcinoma, to make a final conclusion of a certain ratio is very difficult. We envisage that in our group of patients with papillomatosis new cases of carcinoma will occur in the future.

Human papillomavirus in head and neck tumors: epidemiological, molecular and clinical aspects

ZusammenfassungHochrisiko-Typen von humanen Papillomaviren (HR HPV) spielen eine wichtige Rolle in der Ätiologie einer Gruppe von Kopf- und Hals-Plattenepithelkarzinomen. Diese Übersicht konzentriert sich auf die epidemiologischen, molekularen und klinischen Aspekte der HPV-Infektion im Kopf- und Hals-Karzinom. Die DNA von HR HPV wurde in unterschiedlich hohen Anteilen von Kopf- und Halstumoren identifiziert, mit der höchsten Prävalenz im Oropharynx. Patienten mit HPV-assoziierten Tumoren zeichnen sich durch moderaten Tabak- und Alkoholkonsum aus. Manche Aspekte des Sexuallebens können einen Risikofaktor darstellen. Es wurde kürzlich festgestellt, dass sich die HPV-Infektion ausbreitet, und die steigende Prävalenz von HPV-positiven Tumoren ist wahrscheinlich in dieser Epidemie begründet. Auf der molekularen Ebene hat man bewiesen, dass die Onkoproteine E6 und E7 an der Onkogenese beteiligt sind. HPV-positive Karzinome haben eine bessere Prognose, und der HPV-Status sollte bei der klinischen Entscheidung eine Rolle spielen. Der steigende Anteil der HPV-positiven Tumoren unterstreicht die Bedeutung der HPV-Vakzination auch in der Prophylaxe der Kopf- und Halstumoren.SummaryHigh-risk types of human papillomaviruses (HR HPV) play an important role in the etiology of a group of head and neck squamous cell cancers (HNSCC). This review is focused on epidemiological, molecular, and clinical aspects of HPV infection in head and neck cancer. High risk HPV DNA is being detected in a very different proportion of HNSCC with the highest prevalence in oropharynx. Patients with HPV-associated tumors are characterized by moderate tobacco and alcohol consumption. Some aspects of sexual behavior may represent a risk factor. Recently, it has been shown that HPV infection is spreading and the rising prevalence of HPV-positive tumors can probably be attributed to this epidemic. On molecular level the viral oncoproteins E6 and E7 were shown to be involved in oncogenesis. HPV-positive cancers have better prognosis and HPV status should be considered in clinical decision-making. The rising proportion of HPV-positive tumors underlines the importance of HPV vaccination also for the prevention of HNSCC.

Subjective functional results 1 year after surgery and postoperative radiation for oropharyngeal carcinoma

Abstract This study was conducted to find out what factors most influenced functional results and patient satisfaction at 1 year after treatment in a homogeneous group of patients who had undergone surgery and postoperative radiotherapy for the treatment of oropharyngeal carcinoma. Further aims were to find out the relation between the overall and specific disease-related results and to compare the two questionnaires used. The study group consisted of 102 patients, 23 women and 79 men, with an age range of 41–77 years (mean 45.7 years). Two questionnaires were used for evaluation. The University of Washington head and neck quality of life questionnaire (UW-QOL) includes information about nine disease-specific domains and three questions on general quality of life. The second questionnaire used was the performance status scale for head and neck cancer patients (PSS), containing information about three disease-specific domains. Functional results were influenced by the size of tumor and by the surgical method used. Results worsened with increasing T stage. The best results were found after pharyngotomy without mandibulotomy, worse results, following temporary mandibulotomy with osteosynthesis, and the poorest results were found in the group of patients who had undergone composite resection. Neither age and sex of the patients nor the course of healing influenced the functional results. All results were significantly correlated to one another. The general parameters were also closely related to specific domains. An important correlation was found between the parameters in the two questionnaires.

High Level of Tregs Is a Positive Prognostic Marker in Patients with HPV-Positive Oral and Oropharyngeal Squamous Cell Carcinomas

Background. Human papillomaviruses (HPVs) have been proved as one of the etiological factors of oropharyngeal squamous cell carcinoma (OPSCC). Patients with tumors of viral etiology have a lower recurrence rate and better prognosis. OPSCC is linked to an alteration in the immune system. Only a limited number of studies have correlated both the immunological parameters and HPV status with patient prognosis. The aim of this study was to determine whether HPV infection and the immunological status influence patient prognosis individually or in concurrence. Material and Methods. Sixty patients with oral and oropharyngeal carcinomas were enrolled. They were divided into HPV-positive and HPV-negative groups based on the expression of HPV 16 E6 mRNA. Basic lymphocyte subpopulations were determined in the peripheral blood by means of flow cytometry. Results. Significantly better disease-specific survival (DSS) was observed in patients with HPV-positive tumors. Nodal status, tumor grade, recurrence, and CD8+/Tregs ratio were identified as factors influencing DSS. A higher level of Tregs and a lower ratio of CD8/Tregs influenced overall survival (OS) independently of HPV status and age. Patients with HPV-positive tumors and high levels of Tregs survived significantly better than patients from the other groups. Conclusion. Better survival is associated with HPV positivity and elevated Tregs levels. Our data suggest that HPV infection and Tregs do not influence patient prognosis in concurrence.

What are the implications of human papillomavirus status in oropharyngeal tumors for clinical practice

Purpose of reviewHuman papillomavirus (HPV) status itself is an important and very probably the strongest prognostic factor in head and neck cancer. Because of the prognostic advantage of patients with HPV-positive cancers, the issue of the quality of life of survivors has become increasingly important. The possibility of treatment de-escalation in patients with virally induced tumors is being considered. Many challenges have to be addressed in order to integrate HPV status in the routine decision-making in patients with oropharyngeal cancer. The present review discusses the standardization of detection methods suitable for clinical use and the differences in predictive parameters between patients with HPV-positive and HPV-negative tumors. Recent findingsThe gold standard for the identification of patients with oropharyngeal tumors etiologically linked to HPV infection is undoubtedly the detection of HPV 16 E6/E7 mRNA. The detection of a surrogate marker of active viral infection, p16ink4a, has a low sensitivity when used alone and must therefore be combined with the detection of HPV DNA or HPV-specific antibodies. The detailed knowledge of the importance of specific prognostic parameters is crucial in the choice of treatment. Nodal staging is probably much less important in HPV-positive cancers. SummaryIt is of great importance to implement standardized testing for the identification of patients with HPV-induced oropharyngeal tumors. The treatment decision models in HPV-positive tumors have to take into account the probably different prognostic value of nodal parameters. Before introducing treatment de-escalation in patients with virally induced tumors into clinical practice, more research and clinical studies are needed.

Comparison of the miRNA profiles in HPV-positive and HPV-negative tonsillar tumors and a model system of human keratinocyte clones

BackgroundBetter insights into the molecular changes involved in virus-associated and -independent head and neck cancer may advance our knowledge of HNC carcinogenesis and identify critical disease biomarkers. Here we aimed to characterize the expression profiles in a matched set of well-characterized HPV-dependent and HPV-independent tonsillar tumors and equivalent immortalized keratinocyte clones to define potential and clinically relevant biomarkers of HNC of different etiology.MethodsFresh frozen tonsillar cancer tissues were analyzed together with non-malignant tonsillar tissues and compared with cervical tumors and normal cervical tissues. Furthermore, relative miRNAs abundance levels of primary and immortalized human keratinocyte clones were evaluated. The global quantitation of miRNA gene abundance was performed using a TaqMan Low Density Array system. The confirmation of differentially expressed miRNAs was performed on a set of formalin-fixed paraffin-embedded tumor samples enriched for the tumor cell fraction by macrodissection.ResultsWe defined 46 upregulated and 31 downregulated miRNAs characteristic for the HPV-positive tonsillar tumors and 42 upregulated miRNAs and 42 downregulated miRNAs characteristic for HPV-independent tumors. In comparison with the expression profiles in cervical tumors, we defined miR-141-3p, miR-15b-5p, miR-200a-3p, miR-302c-3p, and miR-9-5p as specific for HPV induced malignancies. MiR-335-5p, miR-579-3p, and miR-126-5p were shared by the expression profiles of HPV-positive tonsillar tumors and of the HPV immortalized keratinocyte clones, whereas miR-328-3p, miR-34c-3p, and miR-885-5p were shared by the miRNA profiles of HPV-negative tonsillar tumors and the HPV-negative keratinocytes.ConclusionsWe identified the miRNAs characteristic for HPV-induced tumors and tonsillar tumors of different etiology, and the results were compared with those of the model system. Our report presents the basis for further investigations leading to the identification of clinically relevant diagnostic and/or therapeutic biomarkers for tumors of viral and non-viral etiology.