A subset of sinonasal undifferentiated carcinoma is associated with transcriptionally active high-risk human papillomavirus by in situ hybridization: a clinical and pathologic analysis.

Sinonasal undifferentiated carcinoma (SNUC) is an aggressive malignancy with a poor prognosis, and pathologically, it is a diagnosis of exclusion. Rendering this diagnosis can be challenging in practice because of the large number of diverse entities in the differential diagnosis. We encountered an index case of a sinonasal carcinoma otherwise diagnosable as SNUC which, on further investigation, demonstrated strong and diffuse P16 expression, as well as diffuse expression of high-risk human papillomavirus (hrHPV) RNA by in situ hybridization (ISH). We therefore hypothesized that a subset of cases previously diagnosed as SNUC may in fact harbor transcriptionally active hrHPV. We further investigated a cohort of 25 SNUC cases in our pathology archives and performed ISH for hrHPV RNA on cases that demonstrated >70% nuclear and cytoplasmic P16 expression, criteria which, in other anatomic sites, correlates strongly with the presence of hrHPV. Twelve of 25 SNUC cases were P16 positive, and of these, 5 were positive for hrHPV by ISH. Thus, 20% of all SNUC cases in this cohort harbored transcriptionally active hrHPV. Herein, we report a clinical and pathologic analysis of these cases, including differential diagnostic considerations and comparison of their clinical behavior with SNUC cases that are negative for hrHPV by ISH.

Consistent LEF-1 and MYB Immunohistochemical Expression in Human Papillomavirus-Related Multiphenotypic Sinonasal Carcinoma: A Potential Diagnostic Pitfall.

Human papillomavirus (HPV)-related multiphenotypic sinonasal carcinoma (HMSC) is a distinct, newly-described sinonasal tract neoplasm characterized by a salivary gland tumor-like appearance with myoepithelial and ductal cells, frequent surface squamous dysplasia, and relatively indolent behavior. When considering a diagnosis of HMSC, aggressive high-grade salivary gland carcinomas, particularly those with a basaloid morphology such as basal cell adenocarcinoma and adenoid cystic carcinoma, enter the differential diagnosis. The full morphologic and immunophenotypic profile of HMSC continues to be unraveled. In this series of ten cases, we demonstrate that this tumor has consistent, strong immunohistochemical expression of LEF-1 yet lacks nuclear expression of ß-catenin, and also has consistent yet variable expression of MYB protein. While LEF-1 expression may be a useful diagnostic adjunct, it can also be a pitfall, as other salivary tumors such as basal cell adenocarcinoma have been previously shown to express LEF-1. Additionally, MYB protein expression is not a discriminatory marker when trying to separate HMSC from adenoid cystic carcinoma.

HPV Virus Transcriptional Status Assessment in a Case of Sinonasal Carcinoma.

Human Papilloma Virus (HPV) can play a causative role in the development of sinonasal tract malignancies. In fact, HPV may be the most significant causative agent implicated in sinonasal tumorigenesis and is implicated in as many as 21% of sinonasal carcinomas. To date, there are no definitive, reliable and cost-effective, diagnostic tests approved by the FDA for the unequivocal determination of HPV status in head and neck cancers. We followed an exhaustive algorithm to correctly test HPV infection, including a sequential approach with p16INK4a IHC, viral DNA genotyping and in situ hybridization for E6/E7 mRNA. Here, we report a case of sinonasal carcinoma with discordant results using HPV test assays. The tumor we describe showed an irregular immunoreactivity for p16INK4a, and it tested positive for HPV DNA; nevertheless, it was negative for HR-HPV mRNA. We discuss the possible meaning of this discrepancy. It would be advisable to test HPV transcriptional status of sinonasal carcinoma on a diagnostic routine basis, not only by p16INK4a IHC assay, but also by HPV DNA genotyping and HR-HPV mRNA assessment.

Immunohistochemical Study EMT-Related Proteins in HPV-, and EBV-Negative Patients with Sinonasal Tumours.

Epithelial to mesenchymal transition (EMT) is a biological process in which the epithelial cells, transform to mesenchymal cells via multiple biochemical modifications. Immunohistochemical method was used to examine the expression of EMT-related proteins: Slug, E-cadherin and fibronectin, in 41 cases of sinonasal inverted papilloma (SIP), 33 cases of sinonasal squamous cell carcinoma (SNC), and 22 cases of normal mucosa as a control. In all cases negative viral status was previously confirmed using both in situ hybridization and immunohistochemical method. The immunoexpression of Slug and fibronectin were significantly increased in the SNC group as compared to SIPs and control cases. The immunoexpresssion of Slug was also higher in SIPs as compared to controls. The immunoexpression of E-cadherin was significantly lower in SNCs group as compared with SIPs and controls, but no statistically significant difference in E-cadherin immunoexpression was noted between SIPs and control cases. There were statistically significant negative correlations between immunoexpression of Slug vs E-cadherin, E-cadherin vs fibronectin and positive correlation between Slug vs fibronectin in SNC. Statistically significant correlation between Slug and fibronectin immunoexpression in SIPs was also found. In conclusion, our findings suggest that relationships between Slug, E-cadherin and fibronectin could potentially point to EMT in the sinonasal cancer. Lack of correlation between EMT-related proteins in tested SIPs could reflect a benign nature of those cases.

Treatment outcomes and prognostic factors, including human papillomavirus, for sinonasal undifferentiated carcinoma: a retrospective review.

BACKGROUND: Sinonasal undifferentiated carcinoma (SNUC) is a high-grade, aggressive neoplasm. Low incidence and poor outcomes make identification of prognostic factors and treatment standardization difficult. Similarly, little is known regarding the association of human papillomavirus (HPV) with SNUC. METHODS: A retrospective review was conducted. Extracted information included treatment received, tumor recurrence, patient survival, p16 expression, and HPV status. The Kaplan-Meier method was used to estimate overall survival (OS) and disease-free survival (DFS). Survival trends were compared using the log-rank test. RESULTS: Nineteen patients received multimodality treatment for SNUC. Five-year OS and DFS rates were 45.2% and 50.7%, respectively, with no significant difference between treatment types. Tumors from 11 patients were p16-positive and 9 of these were also HPV-positive. Kaplan-Meier analysis demonstrated improved survival. CONCLUSION: Our series demonstrates a higher prevalence of HPV in SNUC than previously reported. HPV-positive SNUCs may benefit from improved survival and should be investigated further in future studies.

Human papillomavirus load and physical status in sinonasal inverted papilloma and squamous cell carcinoma.

BACKGROUND: This study investigated prospectively the role of human papillomavirus (HPV) in paranasal inverted papilloma (IP). METHODS: HPV presence and viral load and physical status of HPV-16 were examined by polymerase chain reaction-based methods using fresh frozen samples obtained from 13 patients with IP (IP group), 11 with squamous cell carcinoma in the maxillary sinus (SCC group) and 39 with chronic inflammatory lesions (inflammatory group). RESULTS: The presence of the HPV genome was detected in 46.1%, 27.3% and 7.6% of patients in the IP, SCC and inflammatory groups, respectively. The IP group showed significantly higher HPV-positive rates than the inflammatory group. All types of HPV detected were high-risk HPV, especially HPV-16. The relative HPV-16 copy numbers varied from 2.5 to 1524.1 per 50 ng genomic DNA. The viral load was higher in the IP and SCC groups than in the inflammatory group. In the IP group, no significant relationship was found between HPV-16 viral load and clinical characteristics, or between physical status and clinical characteristics. One patient with IP and concomitant squamous cell carcinoma, however, showed high viral load and integration. CONCLUSIONS: HPV infection is involved in the pathogenesis of IP, and high viral load and integration of HPV have an important role in malignant lesion in association with IP.

Significance of human papillomavirus in sinonasal papillomas.

Recent investigations have suggested human papillomavirus (HPV) to be involved in the development of sinonasal papillomas (SNP). Forty-three patients operated for SNP were studied to determine the prevalence of HPV-DNA sequences in these tumours and to evaluate their value as a prognostic parameter. The original sections of all cases were reviewed and reclassified according to the WHO. Paraffin blocks available from 37 patients were subjected to in situ hybridization (ISH) and polymerase chain reaction (PCR). Histology revealed 34 cases of inverted papilloma (IP) (79 per cent), five cases of exophytic papilloma (EP) (12 per cent) and four cases of columnar cell papilloma (CCP) (nine per cent). Recurrences developed in seven of 41 patients (17 per cent), and malignancy occurred in four of 43 patients (nine per cent). HPV was detected in four of 37 specimens (11 per cent) both by ISH and PCR. In particular, HPV-11 was found in three lesions (two EP, one IP) (eight per cent), and HPV-6b was detected in one lesion (one EP) (three per cent). Our findings suggest a possible role for HPV in the pathogenesis of exophytic papillomas. As no correlation was found to malignancy and recurrence of disease, screening for HPV seems not to be useful as a prognostic parameter.

Gamma-rays enhance rAAV-mediated transgene expression and cytocidal effect of AAV-HSVtk/ganciclovir on cancer cells.

Adeno-associated virus (AAV) vector has several unique properties suited for gene therapy applications. However, relatively low efficiency of transgene expression, which is mainly due to a limited second-strand synthesis from the single-stranded AAV genome, can be a problem in some applications that require potent gene expression such as antitumor applications. Recently, gamma-ray irradiation has been reported to enhance the second-strand synthesis of the AAV genome, and consequently transgene expression. We demonstrate here that an AAV vector harboring the herpes simplex virus type-1 thymidine kinase (HSVtk) is able to kill cancer cells more efficiently when used in combination with gamma-ray irradiation. A human maxillary sinus cancer cell line, NKO-1, was efficiently killed in combination with HSVtk transduction and ganciclovir (GCV), as expected. More importantly, gamma-ray irradiation of practical dosages augmented the cytocidal effect of the HSVtk/GCV system. Southern analysis indicated that gamma-rays enhanced the double-strand synthesis of the rAAV genome in NKO-1 cells. These findings suggest that the combination of rAAVtk/GCV suicide gene therapy with radiotherapy has synergistic effects in the treatment of cancers and may lead to a reduction of the potential toxicity of both rAAVtk/GCV and gamma-ray irradiation.

p53 alteration and human papilloma virus infection in paranasal sinus cancer.

BACKGROUND: Inverted papilloma (IP) of the paranasal sinus is a benign neoplastic condition that can be associated with squamous cell carcinoma (SCC). To understand the etiology of the disease better, paranasal sinus tumor specimens were examined for alterations in either p53 protein expression or genomic DNA sequence, and for infection by human papilloma virus (HPV). METHODS: Tumor specimens were categorized as follows: benign, nondysplastic IP; IP with dysplasia; SCC arising within IP; or SCC without IP. Sections of each tumor specimen were stained for p53 protein overexpression, and mutations in exons 5-9 of the p53 gene were determined in DNA purified from all tumor samples. HPV infection was screened by degenerate polymerase chain reaction (PCR) amplification and typed by multiplex PCR and direct DNA sequencing of PCR-amplified HPV sequences. RESULTS: Altered p53, either in genetic sequence or protein overexpression, was observed in 0 of 7 benign, nondysplastic IP specimens. A significantly higher p53 alteration incidence was observed for IP specimens exhibiting dysplasia (57%; P < 0.05) and IP specimens that were associated with SCC (75%; P < 0.025). HPV sequences were detected in 9 of 24 (38%) tumor specimens, 78% of which were of the oncogenic HPV16 strain. A significantly higher incidence (P < 0.05) of HPV infection was observed in IP tumors exhibiting dysplasia or containing SCC than in nondysplastic IPs. None of the p53-mutated tumors were infected with oncogenic HPV16. CONCLUSIONS: These data suggest that p53 alterations and/or HPV infection are associated predominantly with IPs exhibiting evidence of dysplasia or IPs associated with SCC, but not in nondysplastic, benign IPs. In addition, an inverse correlation may exist between oncogenic HPV infection and p53 alterations in paranasal sinus tumors. The authors postulate that patients with IPs containing altered p53 may be at increased risk for SCC of the paranasal sinus.

An expansive papilloma of the nasolachrymal drainage system harbouring human papilloma virus.

We report a case of an expansive tumour extending from the lachrymal sac into the adjacent maxillary sinus. Histology showed a benign exophytic papilloma. By means of the in situ (DNA) hybridization technique, human papilloma virus (HPV) 6/11 were demonstrated, indicating a viral aetiology, similar to exophytic papillomas of the nose and larynx.