HPV-related Tumors Research Articles

The NEDD8 conjugation pathway regulates p53 transcriptional activity and head and neck cancer cell sensitivity to ionizing radiation

Human papillomavirus (HPV)-related oropharyngeal cancer represents a distinct head and neck squamous cell carcinoma (HNSCC) subpopulation, with improved disease-free and overall survival. In general, HPV-positive HNSCCs express wild-type TP53, which could explain its increased radiosensitivity. However, the molecular mechanisms underlying this higher sensitivity remain elusive. We have previously shown that HPV-related oropharyngeal carcinomas express decreased levels of the NEDD8-activating enzyme 1/amyloid β precursor protein-binding protein 1 (NAE1/APP-BP1) gene. NAE1/APP-BP1 function is required for the NEDDylation of target proteins, and has been shown to be a negative regulator of p53 transcriptional activity. In this study, we addressed the hypothesis that NAE1/APP-BP1 expression levels regulate p53 activity and cell survival upon ionizing irradiation. We used the radiosensitive and naturally HPV16-infected UPCI:SCC90 cell line and the radioresistant and HPV-negative SQ20B cell line as the control. NAE1/APP-BP1 expression levels were modulated with expression constructs and siRNAs. Radiosensitivity was evaluated with clonogenic survival assays. p53 transcriptional activity was measured with a luciferase assay. The overexpression of NAE1/APP-BP1 in UPCI:SCC90 cells resulted in the increased NEDDylation of p53, inhibition of p53 activity and increased cell resistance to ionizing radiation. Conversely, the inhibition of NAE1/APP-BP1 expression in SQ20B cells induced p53-dependent cell death after treatment with X-rays. Taken together, these results indicate that NAE1/APP-BP1 and NEDDylation are invovled in modulating p53 activity and regulating its role in the response of cells to ionizing radiation. Our findings bring new insights in the molecular mechanisms underlying the increased radiosensitivity of HPV-related oropharyngeal tumors. This is of importance, as no reliable and robust predictive biomarkers for tumor response to radiotherapy are currently available. These results also have potential clinical significance, as drugs targeting NAE1/APP-BP1 have recently emerged as a novel therapeutic modality in cancer treatment.

Carcinomes épidermoïdes oropharyngés liés à l’infection par les papillomavirus humains de haut risque oncogénique : aspects cliniques, biologiques et perspectives thérapeutiques

The infection of the head and neck epithelium by high-risk human papillomaviruses (HPV) is a risk factor for cancer onset and development. The incidence of HPV-related head and neck squamous cell carcinoma is currently increasing. These lesions display distinct clinical features. HPV positive patients are often younger and have a smaller history of tobacco smoking and alcohol drinking, but have a history of virus-transmitting sex practices. HPV-related tumours are mainly found in the oropharynx, are more associated to a local lymph node invasion and display a poorly differentiated morphology. Despite these more aggressive features, HPV-positive head and neck squamous cell carcinomas correlate with an improved local control, disease-free and global survival. It is thought that HPV-driven specific biologic abnormalities underlie higher tumour sensitivity to chemotherapeutic drugs and ionizing radiations. The expression of the HPV E6 and E7 oncoproteins induce cell transformation by interfering with cell signalling pathways involved in apoptosis, cell cycle, angiogenesis and induce the overexpression of the CDKN2A gene. Therefore, alternative treatments based on therapies targeting these pathways in combination with radiation dose de-escalation could be proposed to HPV-positive patients, if they are properly and reliably identified.

New Concepts for Translational Head and Neck Oncology: Lessons from HPV-Related Oropharyngeal Squamous Cell Carcinomas

Human papillomavirus (HPV) infection is well established as an etiological agent responsible for a number of pathologies affecting the stratified epithelia of skin and anogenital sites. More recently, the infection by (mucosal) high-risk HPV types has also been found to be causally associated with squamous cell carcinoma in the head and neck region (HNSCC), especially in the oropharynx. Intriguingly, HPV-related oropharyngeal squamous cell carcinomas (OPSCC) represent a distinct clinical entity compared to HPV-negative tumors with particular regard to treatment–response and survival outcome. The association between HPV infection and OPSCC may therefore have important implications for the prevention and/or treatment of OPSCC. The improved survival of patients with HPV-related tumors also raises the question, as to whether a better understanding of the underlying differences may help to identify new therapeutic concepts that could be used in targeted therapy for HPV-negative and improved therapy for HPV-positive cancers. This review summarizes the most recent advances in our understanding of the molecular principles of HPV-related OPSCC, mainly based on functional genomic approaches, but also emphasizes the significant role played by the tumor microenvironment, especially the immune system, for improved clinical outcome and differential sensitivity of HPV-related tumors to current treatment options.

Epidemiologic profile, sexual history, pathologic features, and human papillomavirus status of 103 patients with penile carcinoma

The incidence of penile cancer is four times higher in Paraguay than in the United States or Europe. There are no adequate scientific explanations for this geographical variation. The goal of this study was to evaluate the interplay among risk factors, morphology of the primary tumor, and HPV status. Information on socioeconomic status, education level, habits, and sexual history was obtained in 103 Paraguayan patients with penile cancer. All patients were then treated by surgery, and specimens were evaluated histopathologically. Patients usually dwelled in rural/suburban areas (82%), lived in poverty (75%), had a low education level (91%), and were heavy smokers (76%). Phimosis (57%), moderate/poor hygienic habits (90%), and history of sexually transmitted diseases (74%) were frequently found. Patients with >10 lifetime female partners had an odds ratio of 3.8 (95% CI 1.1, 12.6; P-trend = .03) for presenting HPV-positive tumors when compared to patients with <6 partners. However, this trend was not significant when the number of sexual partners was adjusted for age of first coitus and antecedents of sexually transmitted diseases. HPV-related tumors (found in 36% of the samples) were characterized by a warty and/or basaloid morphology and high histological grade in most cases. In our series, patients with penile cancer presented a distinctive epidemiologic and pathologic profile. These data might help explaining the geographical differences in incidence and aid in the design of strategies for cancer control in Paraguay.

Next‐generation treatment strategies for human papillomavirus‐related head and neck squamous cell carcinoma: where do we go?

Oncogenic human papillomavirus (HPV) is currently recognised as a major risk factor for the development of head and neck squamous cell carcinomas (HNSCC). HPV is mostly detected in tumours arising from the oropharynx and more specifically from the tonsil. HPV-related tumours display clinical and molecular characteristics that are distinct from HPV-unrelated tumours, which are generally induced by alcohol and tobacco abuse. Detection of biologically active HPV in HNSCC has prognostic relevance, which warrants the separate classification of HPV-induced tumours and is a prerequisite for further optimisation of treatment protocols for this distinct group. Current guidelines for the treatment of oropharyngeal squamous cell carcinoma (OPSCC) have not incorporated specific treatment modalities for HPV-related tumours. The development of such treatment options is still in a preclinical phase or in early clinical trials. Recent data on treatment response of OPSCC have been obtained by retrospectively analysing HPV-status and indicate that patients with HPV-related tumours show a favourable prognosis, independent of the type of treatment. These patients may benefit from de-intensified treatment, which should be assessed in prospective clinical trials. The development and future use of new antiviral and immunomodulatory therapeutics may be instrumental in this approach to improve survival rates and decrease disease-and-treatment-related morbidity. In this review we will focus on present therapeutic HPV-targeting strategies and discuss future directions for de-intensified treatment of HPV-positive HNSCC.

The role of cytopathology in diagnosing HPV induced oropharyngeal lesions

HPV detection in fine needle aspirates from suspected head and neck metastasis may be useful in clinching the diagnosis of HPV related oral squamous cell carcinoma. Ascertaining the HPV status of a particular tumor on cytological specimens could be useful for prognostication as HPV-related tumors appear to have a better prognosis and clinical outcome. The various techniques of detection are reviewed.

Abstract 5061: Differentially expressed microRNA signatures based on human papillomavirus status in head and neck squamous cell carcinoma

Abstract Introduction: Recent studies have demonstrated that human papillomavirus (HPV) status is a favorable prognostic factor in patients with head and neck squamous cell carcinoma (HNSCC). Additionally, microRNAs (miRNAs) have emerged as an important regulator of global gene expression, and their signaling is often altered in cancer. Thus, we assessed the differences in global miRNA expression in normal mucosal epithelia (NME), HPV-positive (+), and HPV-negative (−) tumors to gain insight into patient prognosis given the current standard of care. An understanding of these mechanistic differences could lead to potential therapeutic targets and improvement in clinical outcomes for patients with HPV (−) tumors. Methods: miRNAs were isolated from 6 NME and 20 frozen tumors using PureLink RNA Isolation Kit, and miRNA using PureLink microRNA Isolation Kit (Invitrogen). miRNA was analyzed using the ABI Megaplex protocol without pre-amplification (Applied Biosystems). Reactions were run using the ABI miRNA Reverse Transcription Kit and Megaplex RT Human Pool A primers, then loaded onto TaqMan Low-Density Array (TLDA) cards. TLDAs were analyzed using an ABI7900HT Real Time PCR machine using the default ABI TLDA protocol. A tissue microarray (TMA) of 153 cores from an additional 51 HNSCC tumors was stained with a phospho-specific AKT (pAKT) antibody (Ser473, Cell Signaling Technology), and digitally quantified with TMAJ software (Johns Hopkins Univ.). After cores were omitted due to lack of tumor or unknown HPV-status, staining was analyzed from 123 cores from 45 tumors and evaluated using a two-sample t-test assuming equal variances. Results: The number of differentially expressed miRNAs were as follows: 36 in HPV (+) compared to NME; 29 in HPV (−) compared to NME; 40 shared between the tumor sets compared to NME; and one (miR-449a) between both tumor sets after multi-variable corrections. An miRNA lower in both HPV (+) and HPV (−) tumors with respect to NME was miR-124, a putative regulator of PIK3CA. To contrast functional differences in PI3KCA signaling with HPV status, a TMA of 45 HNSCC tumors was examined by IHC for pAKT expression. The mean pAKT staining was higher in HPV (+) tumors compared to HPV (−) cohorts (p = 0.003). Interestingly, lower pAKT levels correlated with tumor recurrence in HPV (+) tumors (p = 2.5×10−8), while no statistical difference was noted in HPV (−) tumors (p = 0.24). Conclusions: There are significant differences in the miRNA profiles of tumors compared to NME, suggesting deregulation of miRNAs may play an important role in the development of HNSCC. Our data suggest the downstream effects of miR-124 regulation may differ in the context of HPV status, and that PI3K/AKT signaling may influence the observed difference in HPV-related tumor clinical outcomes. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5061. doi:10.1158/1538-7445.AM2011-5061

Assessment of Immunohistochemistry for p16INK4 and High-Risk HPV DNA by In Situ Hybridization in Esophageal Squamous Cell Carcinoma

The role of high-risk human papillomavirus (HPV) in the pathogenesis of esophageal squamous cell carcinoma (ESCC) remains unclear. p16(INK4) is used as a surrogate marker to detect HPV-related tumors but has had discrepant results in ESCC. In this study, 32 cases of ESCC were examined to determine the relationship between p16(INK4) expression and high-risk HPV. All the tumors were stained by immunohistochemistry for p16(INK4). Tumors having p16(INK4) nuclear and/or nuclear and cytoplasmic expression were considered positive. Tumors positive for p16(INK4) expression were tested for high-risk HPV by in situ hybridization (ISH). In all, 20 cases of ESCC (63%) showed only cytoplasmic staining for p16(INK4), and 11 cases (34%) showed both cytoplasmic and nuclear staining for p16(INK4); 4 cases (13%) showed no staining for p16(INK4). None of the p16(INK4) -positive cases were positive for high-risk HPV by ISH. These results indicate that p16(INK4) expression in ESCC does not correlate with the presence of high-risk HPV DNA by ISH. High-risk HPV does not seem to play a major role in the carcinogenesis of ESCC in low-risk areas.

Head and Neck Pathology: New Developments in the Diagnosis and Pathogenesis of Head and Neck Tumors

In this issue manuscripts are presented that touch on some of the new developments in the diagnosis and pathogenesis of head and neck tumors. The first article by T. Tanaka et al. summarizes our understanding of the development of oral squamous cell carcinoma and discusses possible chemoprevention. As discussed there, the diagnosis of head and neck cancer can be limited as the entirety of the upper aerodigestive tract can be affected by smoking. P. Pujary et al. discuss the use of spectroscopy for the identification of malignant squamous mucosa. As the incidence of smoking decreases in many parts of the world, there has been a noted decline in the incidence of smoking-related malignancies, including head and neck squamous cell carcinoma. Unfortunately, with this decline, many parts of the world have witnessed a concomitant increase in oropharyngeal squamous cell carcinomas related to high-risk human papillomavirus (HPV) infection. In this issue, Dr. F. Farshadpour et al. show, in a case control study, that oropharyngeal squamous cell carcinomas that develop in nonsmokers who do not abuse alcohol are more likely to be related to HPV infection than those that develop in smokers who do consume alcohol and confirm that the HPV-related tumors have a better prognosis. R. L. Cantley et al. provide a nice review of the various immunohistochemical, in situ hybridization, and molecular studies that can be used to demonstrate HPV infection. Over the past decade there have been many exciting developments in our understanding of the classification of salivary gland neoplasms and the associated genetic abnormalities. In this issue, M. Shishegar et al. describe a series of salivary gland tumors seen at their institution over a 6-year period. A. F. Costa et al. review the clinicopathologic changes associated with high-grade transformation that can be seen with adenoid cystic carcinomas, acinic cell carcinomas, epithelial-myoepithelial carcinomas, and polymorphous low-grade adenocarcinomas. Sinonasal intestinal-type adenocarcinomas remain a pathologic enigma, sometimes associated with heavy wood-dust exposure. In the final article in this issue, B. Vivanco et al. review a large series of these tumors and attempt to identify possible precursor lesions for these rare malignancies. Stefan Pambuccian Edward B. Stelow Ioannis G. Koutlas Michael J. Thrall

Randomized Clinical Trial on Postoperative Continuous 7-days-a-week Radiotherapy vs. Postoperative Radiochemotherapy in Squamous-cell Head and Neck Cancer: A Report on Acute Normal Tissue Reactions

Conclusions: This study identified a population of patients with multiple potentially HPV-related tumors and demonstrates differ- ent outcomes based on multiple factors, including the site of first diagnosis and duration between diagnoses. Interval between diagnoses was also found to depend upon the site of the initial tumor. In patients presenting with a potentially HPV-related ma- lignancy,anopportunityforsecondarycancerpreventionmayexistthroughbehaviormodificationandalteredscreening, butfuture work is needed to establish true HPV-positivity in this cohort and to understand a potential role for vaccine and chemoprevention strategies. Results: All patients in p-CAIR completed radiotherapy as prescribed, with no need for unplanned interruptions. By contrast, only 59% of the patients in p-CRT received 3 courses of chemotherapy, while cisplatin was discontinued after 2 courses in 37%, and after thefirst coursein4%. Two patientsinp-CRT hadradiation treatmentinterruptions of7 days ormore.Severeadverseeventsin p-CRT included one patient that died due to sepsis (2.4%), one patient with pulmonary embolism (2.4%) and 5 cases of neurotox- icity (12.1%). In addition, 32% of the patients in p-CRT had grade 3 neutropenia, compared to 0% in p-CAIR (p\0.001). The average hemoglobin concentration at the end of radiotherapy was significantly lower in p-CRT compared to p-CAIR (13.5 g/dl vs. 12.0 g/dl, p = 0.02. The platelets count at the end of radiotherapy was lower in p-CRT vs. p-CAIR, but the difference was not sig- nificant (250*10 5 vs. 210*10 5 , p = 0.12). The average maximum total severity score for mucosal reactions was significantly higher in patients treated with p-CAIR compared to p-CRT (15.3 vs. 13.0, p = 0.03). The average duration of severe mucosal reactions (score $10) did not, however, differ significantly in the respective arms (2.9 weeks p-CAIR vs. 2.6 weeks p-CRT). Conclusions: The early tolerance of p-RCT was considerably affected by hematological and neurological reactions that necessi- tated discontinuation of chemotherapy in 41% of the patients. The maximal intensity of acute mucosal reactions was higher in p- CAIR compared to p-RCT, but the duration of severe reactions did not differ significantly between the arms.

Human papillomavirus 16 E6 increases the radiosensitivity of p53-mutated cervical cancer cells, associated with up-regulation of aurora A

Purpose: To examine the effect of the human papillomavirus (HPV) type 16-E6 (HPV ‘early’ gene) oncoprotein on in vitro radiosensitivity of HPV-negative/p53 mutant C33a cervical cancer cells.Methods and materials: The human cervical cancer cell line C33a was stably transfected with either the HPV16 E6 cDNA cloned into the vector pcDNA™3.0 (C33aE6) or the empty-vector control (C33aV). Radiosensitivity, DNA damage, and cell cycle measurements were made using standard clonogenic assays, immunofluorescent assessment of nuclear histone H2AX phosphorylated on serine-139 (γ-H2AX) foci, and flow cytometry. Western immunoblotting and fluorescence confocal microscopy were used to analyse the changes in cellular proteins. Real-time polymerase chain reaction (PCR) was used to compare levels of aurora A mRNA.Results: Compared to C33aV cells, C33aE6 cells showed enhanced radiation cell killing. This was associated with a large amount of polyploidy which was followed by late cell death in C33aE6 cells. Aurora A was highly expressed in C33aE6 cells at pre- and post-irradiation times compared to C33aV cells. Silencing aurora A resulted in a reduced amount of residual γ-H2AX foci in C33aE6 cells, and diminished the difference in radiosensitivity between the C33aE6 and C33aV cells.Conclusion: Our in vitro results indicate that genetic instability could be augmented in the HPV-infected cancer cells by up-regulation of aurora A, especially against a background of dysfunctional p53. Further studies are needed to examine whether aurora A could be a viable therapeutic target in HPV-related tumours.

Plasma human papillomavirus (HPV) DNA as a potential tool for tumor detection and monitoring response in HPV-related oropharyngeal carcinoma (OP).

5529 Background: There is a rising incidence of HPV-related OP, which has a distinct prognosis. Previous studies in other viral-related cancers such as nasopharyngeal carcinoma have shown that the level of circulating instigating virus DNA in the blood can be used to monitor treatment response and tumor relapse. The main objective of this study is to develop a similar assay for HPV(+) OP. Methods: We designed a semiquantitative PCR approach to detect circulating HPV DNA using the L1 Gp5/6 degenerate primer as well as HPV16/18 E6 and E7 nested primers. We used this approach to detect HPV DNA in culture media of SCC90, a HPV(+) OP cell line as well as plasma samples of SCC90 bearing SCID mice. Control mice were those implanted with non-HPV tumors. HPV DNA was also assessed in pretreatment plasma samples of 32 HPV(+) OP patients and 4 HPV(-) control patients with other head and neck squamous cell carcinoma. HPV status within the tumor was confirmed by both p16 staining and HPV16/18 PCR (N = 30) or in-situ hybridization (n = 2). Plasma HPV DNA levels were also quantified by a fluorescent probe multiplex real-time PCR (QRTPCR) assay, targeting the HPV16/18 E6 region, in 6 patients with HPV(+) OP and serial plasma samples obtained during radiotherapy (RT). Results: Cell-free HPV DNA was detected in the culture media of SCC90 and the plasma of SCC90 bearing mice but not in control mice. It was detected in pretreatment plasma of 84% HPV(+) patients and in none of the HPV(-) controls using the L1 primers and conventional PCR. However using less sensitive primer sets (nested E6 and E7), plasma HPV DNA was positive in only 50% of patients with detectable E6/E7 DNA in the tumors. QRTPCR was able to detect as low as 10 copies of HPV DNA in each reaction. In 6 patients with serial plasma samples during RT, QRTPCR measurement showed a rapid decline in HPV DNA level, which became undetectable at RT completion. Conclusions: The xenograft studies indicate that plasma HPV DNA is released directly from HPV-related tumors. The high sensitivity of this method allows detection of HPV DNA in plasma even with low viral load. Plasma HPV DNA may be a valuable tool for monitoring therapeutic response and disease progression in HPV(+) OP. No significant financial relationships to disclose.

Comparison of Subtypes of Penile Squamous Cell Carcinoma From High and Low Incidence Geographical Regions

There is a worldwide geographical variation in the incidence of penile squamous cell carcinoma (PSCC); some subtypes are HPV-related (warty, basaloid) while others (keratinizing variants) are not. The aims of this study were to compare the distribution of different histological subtypes of PSCC from regions of low (Paraguay, 144 cases) and high (USA, 157 cases) incidence and to identify and compare tumors with and without warty and/or basaloid morphology. The distribution of subtypes in the Paraguayan and the American series was: usual, 49.3 and 46.5%; verrucous, 8.3 and 7.6%; papillary NOS, 7.6 and 5.7%; warty, 6.9 and 8.3%; basaloid, 4.2 and 7.0%; sarcomatoid, 0.7 and 0.6%; adenosquamous, 3.5 and 0.6%; and mixed, 19.4 and 23.6%, respectively. The distribution of mixed PSCC was: warty-basaloid, 50.0 and 59.5%; usual-verrucous, 21.4 and 21.6%; usual-warty, 14.3 and 8.1%; usual-basaloid, 7.1 and 0.0%; usual-papillary, 3.6 and 5.4%; and others, 3.6 and 5.4%, respectively. In conclusion, we found no geographical difference in the incidence of histological subtypes (p = 0.6501), mixed PSCC (p = 0.5937) or HPV-related tumors (p = 0.2505). Geographical variation may be the result of staging variation at clinical presentation or of pathological diagnosis. The identification of similar histological subtypes in both series validates this classification approach for penile cancer. The tendency for typical SCC to mix with verrucous and papillary carcinomas and of the basaloid to preferentially mix with benign condyloma and condylomatous (warty) carcinomas would support the hypothesis of the existence of an etiologically different dual population of penile tumors.

Synchronous multifocal HPV-related neoplasm involving both the genital tract and the head-and-neck area: A case report of Fanconi anemia

We report the case of a 32-year-old woman who presented with multiple squamous cell neoplasms synchronously arising in the cervix, vulva, oral cavity and oropharynx. During the period when the patient was treated by radiotherapy, she developed severe radiation mucositis and pancytopenia. Immunofluorescence staining for foci of the monoubiquitinated form of FANCD2 revealed a complete absence of such foci in all cancer tissues that were examined.

Controversies surrounding human papilloma virus infection, head &amp; neck vs oral cancer, implications for prophylaxis and treatment

Head & Neck Cancer (HNC) represents the sixth most common malignancy worldwide and it is historically linked to well-known behavioural risk factors, i.e., tobacco smoking and/or the alcohol consumption. Recently, substantial evidence has been mounting that Human Papillomavirus (HPV) infection is playing an increasing important role in oral cancer. Because of the attention and clamor surrounding oral HPV infection and related cancers, as well as the use of HPV prophylactic vaccines, in this invited perspective the authors raise some questions and review some controversial issues on HPV infection and its role in HNC, with a particular focus on oral squamous cell carcinoma.The problematic definition and classification of HNC will be discussed, together with the characteristics of oral infection with oncogenic HPV types, the frequency of HPV DNA detection in HNC, the location of HPV-related tumours, the severity and prognosis of HPV-positive HNC, the diagnosis of oral HPV infection, common routes of oral infection and the likelihood of oro-genital HPV transmission, the prevention of HPV infection and novel therapeutic approaches.

Construction and characterization of recombinant fowlpox viruses expressing human papilloma virus E6 and E7 oncoproteins

Human papilloma virus (HPV)-16 is the most prevalent high-risk mucosal genotype and the expression of the E6 and E7 proteins, which can bind to the p53 and p105Rb host cell-cycle regulatory proteins, is related to its tumorigenicity. Virus-like-particle (VLP)-based immunogens developed recently are successful as prophylactic HPV vaccines. However, given the high number of individuals infected already with HPV and the absence of expression of the L1 structural protein in HPV-infected or HPV-transformed cells, an efficient therapeutic vaccine targeting the non-structural E6 and E7 oncoproteins is required. In this study, two new fowlpox virus (FPV) recombinants encoding the HPV-16 E6 and E7 proteins were engineered and evaluated for their correct expression in vitro, with the final aim of developing a therapeutic vaccine against HPV-related cervical tumors. Although vaccinia viruses expressing the HPV-16 and HPV-18 E6 and E7 oncoproteins have already been studied, due to their natural host-range restriction to avian species and their ability to elicit a complete immune response, FPV recombinants may represent efficient and safer vectors also for immunocompromised hosts. The results indicate that FPV recombinants can express correctly the E6 and E7 oncoproteins, and they should represent appropriate vectors for the expression of these oncoproteins in human cells.

Detection of human papillomavirus-related squamous cell carcinoma cytologically and by in situ hybridization in fine-needle aspiration biopsies of cervical metastasis

Fine-needle aspiration (FNA) biopsy often is the first diagnostic procedure performed in patients with head and neck masses. When squamous cell carcinoma (SCC) is diagnosed, proper management and improved prognosis depends on identification of the primary tumor. Recent studies have indicated that human papillomavirus (HPV) infection is associated closely with oropharyngeal SCC and that these tumors have a distinct nonkeratinizing morphology. In this study, the authors explored the value of identifying HPV-related tumors in neck metastases to determine the origin of occult primary head and neck squamous cell carcinoma (HNSCC). Thirty FNA biopsies of neck metastases from patients with HNSCC were recovered from the authors' files from 2004 to 2005. The primary sites included 13 oropharynx, 13 oral cavity, and 4 larynx/hypopharynx. All patients had corresponding tissue samples from the neck mass and the primary carcinoma. The FNA specimens and corresponding tissue samples were classified as either nonkeratinizing SCC (NKSCC) or keratinizing SCC (KSCC). In situ hybridization for HPV (HPV-ISH) was performed using ethanol-fixed, Papanicolaou-stained smears. A positive signal was defined as dark blue or black nuclear dots. Corresponding formalin-fixed, paraffin-embedded tissue sections also were processed for HPV-ISH. Twenty of the 30 FNA specimens were KSCC, and 10 were NKSCC. Eight of the 10 NKSCCs originated in the oropharynx, and 2 had nonoropharyngeal origin. HPV was detected in 7 of 10 NKSCCs. Ten of 30 (33%) FNA biopsies were positive for HPV, and 9 of those biopsies were metastatic from the oropharynx. Nonkeratinzing morphology or HPV-positive ISH in FNA samples significantly predicted oropharyngeal origin (P < .0069 and P < .0004, respectively). NKSCC in metastatic cervical lymph nodes predicted positive HPV-ISH and was strongly suggestive of an oropharyngeal primary tumor.

Human papillomavirus seropositivity and risks of head and neck cancer

We examined antibody response to VLP HPV-16, HPV-16 E6 and E7 antibodies as potential seromarkers of HPV-related head and neck cancer (HNC). The study included 204 HNC cases and 326 controls evaluated for HPV presence in sera using ELISAs for anti-HPV VLP antibodies and HPV-16 E6 and/or E7 antibodies, and in tumor tissue using PCR and DNA sequencing. Anti-HPV-16 VLP was detected in 33.8% of cases and 22.4% of controls, anti-E6 in 20.6% of cases and 0.9% of controls and anti-E7 in 18.6% of cases and 0.6% of controls. HPV-16 DNA was detected in 26.1% of tumors. The adjusted risk of HNC was elevated among those seropositive for HPV-16 VLP (odds ratio (OR) = 1.7, 1.1-2.5), E6 (OR = 32.8, 9.7-110.8) or E7 (OR = 37.5, 8.7-161.2). Compared to HPV DNA-negative/seronegative cases, tumor HPV-16 cases had increased risk of detection with anti-VLP antibodies (OR = 6.8, 3.1-14.9). The odds were more pronounced among cases seropositive for E6 (OR = 69.0, 19.3-247) or E7 (OR = 50.1, 14.7-171). Antibodies against E6 or E7 were associated with risk of cancer in the oral cavity (OR = 5.1, 1.2-22.4) and oropharynx (OR = 72.8, 16.0-330), and with disease characteristics: stage, grade and nodal status. Anti-E6 and/or E7 antibodies were found in 74% of tumor HPV-16 positive cases but in only 5% of tumor HPV-negative cases (K =0.7, 0.6-0.8) suggesting good correlation between the serologic marker and HPV tumor status. Antibodies to HPV-16 E6 and/or E7 represent a more specific biomarker than anti-HPV-16 VLP of an HPV-related HNC. Because of the survival advantage of HPV-related HNC, HPV-16 E6/E7 detection may be useful in therapy targeted for HPV-related tumors.

Estimating the long-term impact of a prophylactic human papillomavirus 16/18 vaccine on the burden of cervical cancer in the UK

To predict the public health impact on cervical disease by introducing human papillomavirus (HPV) vaccination in the United Kingdom, we developed a mathematical model that can be used to reflect the impact of vaccination in different countries with existing screening programmes. Its use is discussed in the context of the United Kingdom. The model was calibrated with published data. The impact of vaccination on cervical cancer and deaths, precancerous lesions and screening outcomes were estimated for a vaccinated cohort of 12-year-old girls, among which it is estimated that there would be a reduction of 66% in the prevalence of high-grade precancerous lesions and a 76% reduction in cervical cancer deaths. Estimates for various other measures of the population effects of vaccination are also presented. We concluded that it is feasible to forecast the potential effects of HPV vaccination in the context of an existing national screening programme. Results suggest a sizable reduction in the incidence of cervical cancer and related deaths. Areas for future research include investigation of the beneficial effects of HPV vaccination on infection transmission and epidemic dynamics, as well as HPV-related neoplasms in other sites.

Carcinomas of the Female Genital Tract Occurring After Pelvic Irradiation

The concept of postradiation sarcoma is widely appreciated, however carcinomas arising in previously irradiated fields, the putative "postradiation carcinoma," are less well understood. Fifteen patients who developed gynecological malignancies after pelvic radiation therapy were studied. Five of these patients had HPV-related tumors both pre- and post- irradiation. Ten were irradiated for cervical cancer, one for endometrial carcinoma, one for vulvar carcinoma, one for colon cancer and 2 for benign conditions. The mean and median latent periods from the initiation of radiation therapy to the development of the second malignancy were 22.8 and 19 years, respectively (22.4 and 19.5 years, respectively, for non-HPV-related cancers; 24 and 18 years for HPV-related cancers). The "postradiation" malignancies included 2 ovarian carcinomas, 5 vaginal carcinomas (3 invasive, 2 in situ), 4 endometrial carcinomas, one cervical carcinoma, one vulvar carcinoma, one distal urethral carcinoma, and one pelvic carcinoma of unclear primary site. Gynecological carcinomas may occur many years after pelvic irradiation. Although the evidence for a causative role is circumstantial, these tumors appear to have a similar latent period as postradiation sarcomas.