Daniel Weiss

Prevalence and impact on clinicopathological characteristics of human papillomavirus‐16 DNA in cervical lymph node metastases of head and neck squamous cell carcinoma

Human papillomavirus (HPV) is a basic risk factor for head and neck squamous cell carcinoma (HNSCC). Little knowledge exists about the impact of HPV on clinical diagnostic and therapy of patients with HNSCC.

Promoter methylation of cyclin A1 is associated with human papillomavirus 16 induced head and neck squamous cell carcinoma independently of p53 mutation

Aberrant promoter methylation of specific genes and infection with human papillomavirus 16 (HPV16) are known risk factors for the development of Head and Neck Squamous Cell Carcinoma (HNSCC). Little knowledge exists on the interaction of HPV16 infection and promoter methylation in HNSCC. The promoter methylation status of 12 genes (TIMP3, CDH1, CDKN2A, DAPK1, transcription factor 21 (TCF21), CD44, MLH1, MGMT, RASSF1, cyclin A1 (CCNA1), LARS2, and CEBPA) was evaluated by methylation‐specific polymerase chain reaction in 55 primary HNSCC and 31 controls. The results were correlated with HPV16 status and clinicopathological characteristics. CCNA1 and p53 protein expression were additionally determined by immunohistochemistry and compared with p53 mutation status. Methylation of DAPK1 (P = 0.043), CCNA1 (P = 0.016) and TCF21 (P = 0.0005) was significantly more present in HNSCC than in controls. The genes TIMP3 (P = 0.018) and CCNA1 (P = 0.015) showed higher methylation frequency in HPV16 positive HNSCC compared to HPV16 negative tumors. CCNA1 methylation did not correlate with CCNA1 protein expression and p53 mutation, respectively. Methylation of TCF21 was associated with higher age (P = 0.044) and nicotine abuse (P = 0.035). Methylation of CCNA1 was significantly more present in females (P = 0.003). Methylation of TCF21 and CCNA1 are important risk factors for HNSCC development. CCNA1 methylation may play a crucial role in HPV16‐induced carcinogenesis of HNSCC independently of p53.

Protein expression and promoter methylation of the candidate biomarker TCF21 in head and neck squamous cell carcinoma

PurposeEpigenetic alterations of the transcription factor 21 (TCF21) gene have been associated with head and neck squamous cell carcinoma (HNSCC) and other tumor entities. So far, however, no reports have appeared in the literature on TCF21 protein expression in HNSCC and its relevance as a putative biomarker.MethodsTCF21 protein expression was assessed in 74 HNSCCs and 31 benign tonsils by immunohistochemistry. Methylation analyses of the corresponding gene promoter were performed in 45 HNSCCs and 31 benign tonsils. The TCF21 expression levels in the tumors and controls were compared with each other and within each group and, in addition, with the TCF21 promoter methylation status and various clinicopathological characteristics.ResultsOverall, both the expression levels and methylation frequencies of TCF21 were significantly higher in the HNSCCs than in the benign controls (p < 0.001 each). Specifically, TCF21 promoter hypermethylation resulted in a reduced protein expression in a subgroup of the HNSCCs (p = 0.038), but not in the tonsils. In the tonsils, TCF21 protein expression positively correlated with that of CD31 (p = 0.039), a marker for blood vessels. Also, in the tonsils the TCF21 protein methylation frequency showed a positive correlation with age (p = 0.008). The HNSCCs of patients with a positive history for alcohol and nicotine abuse showed higher TCF21 protein expression levels than their respective counterparts (p = 0.028 and p = 0.062, respectively). The same was observed in human papilloma virus (HPV)-negative tumors (p = 0.042), tumors located in the oral cavity (p = 0.016) and early-stage tumors (p = 0.025). Interestingly, expression rates in tumors of the oropharynx, where HPV-positive tumors were most frequently found, tended to be lower (p = 0.065). The methylation frequencies of TCF21 were found to be significantly higher in tumors of patients without nicotine abuse (p = 0.030), in HPV-positive tumors (p = 0.014) and in tumors exhibiting over-expression of p16, a protein induced by HPV (p = 0.006).ConclusionsBoth over-expression and increased promoter methylation of TCF21 were frequently observed in HNSCCs. TCF21 promoter hypermethylation was found to lead to gene silencing in the HNSCCs, but not in the benign tonsils. These epigenetic, and possibly also genetic, alterations of the TCF21 gene in HNSCCs may be driven by HPV infection, nicotine and alcohol abuse, or both. These findings, together with its stage- and primary site-dependent expression, turn TCF21 into a promising candidate biomarker in HNSCC.

Platelet Glycoproteins and Fibrinogen in Recovery from Idiopathic Sudden Hearing Loss

Background The pathomechanism and location of idiopathic sudden sensorineural hearing loss (ISSHL) is unclear. In a previous case-control study, we found elevated fibrinogen concentrations and a higher prevalence of T allele carriers of the glycoprotein (Gp) Ia C807T polymorphism in ISSHL patients. Methodology 127 patients with ISSHL (mean age 53.3 years, 48.8% females), who underwent a standard therapy with high dose steroids, pentoxifyllin and sterofundine over 8 days were included. We examined the influence of GpIa genotype and fibrinogen (BclI-, A312-, HaeIII-) genotype and fibrinogen plasma levels on hearing recovery after 8 weeks (change from baseline: 0 dB  =  no recovery, >0 to 10 dB = moderate recovery, >10 dB = good recovery). In a subsample of 59 patients with ISSHL, we further studied the association of platelet glycoprotein GpIa, Ib and IIIa densities on hearing recovery as well as the possible effect-modification of platelet glycoproteins on hearing recovery by plasma fibrinogen. Results In univariate analysis, neither the GpIa genotype nor fibrinogen genotype (all p>0.1) but lower fibrinogen levels (p = 0.029), less vertigo (p = 0.002) and lower GpIIIa receptor density (p = 0.037, n = 59) were associated with hearing recovery. In multivariate analysis, fibrinogen significantly modified the effect of GPIa receptor density on good hearing recovery (effect-modification on multiplicative scale OR = 0.45 (95% confidence interval (0.21–0.94)), p = 0.03). GPIb receptor density below the mean was associated with a 2-fold increase in good hearing recovery both in patients with fibrinogen levels above (p = 0.04) as well as in patients with fibrinogen levels below the mean (p = 0.06). There was no indication for an effect-modification (p = 0.97). Conclusions The findings suggest a vascular/rheological origin of ISSHL with unique features of thrombosis in the inner ear artery that may include complex interrelationships among platelet glycoproteins and plasma fibrinogen.

Cyclin A1 shows age-related expression in benign tonsils, HPV16-dependent overexpression in HNSCC and predicts lower recurrence rate in HNSCC independently of HPV16

BackgroundPromoter methylation of the tumor suppressor gene Cyclin A1 could be associated with Human Papillomavirus 16 (HPV16) induced Head and Neck Squamous Cell Carcinoma (HNSCC) and Cervical Carcinoma. There is disagreement about the impact of this epigenetic event on protein expression of Cyclin A1 in malignant and non-malignant tissue and there hardly exists any information about possible relationships between Cyclin A1 expression and clinicopathological characteristics in HNSCC.MethodsWe analyzed protein expression of Cyclin A1 in 81 HNSCC and 74 benign tonsils by immunohistochemistry and correlated it to Cyclin A1 methylation status, presence of HPV16 infection and other clinicopathological characteristics.ResultsOverexpression of Cyclin A1 was more present in HNSCC than in tonsils (p < 0.001). In both entities, HNSCC and benign tonsils, expression of Cyclin A1 significantly correlated with the expression of Cyclin-dependent kinase-inhibitor p16 (p = 0.000672 and 0.00495). In tonsils, expression of Cyclin A1 was inversely proportional to age (p = 0.00000396), and further correlated with expression of tumor suppressor gene p53 (p = 0.000228). In HNSCC Cyclin A1 expression was associated with the presence of HPV16 DNA (p = 0.0014) and a lower recurrence rate in univariate and multivariate analysis (p = 0.002 and 0.013). Neither in HNSCC nor in tonsils Cyclin A1 expression correlated with promoter methylation.ConclusionsCyclin A1 is an important cell cycle regulator with age-related increased expression in tonsils of children. HPV16 induces overexpression of Cyclin A1 in HNSCC despite promoter methylation. Overexpression of Cyclin A1 predicts a lower recurrence rate in HNSCC independently of HPV16.

Exploratory study of long-term health-related quality of life in patients with surgically treated primary parotid gland cancer

Health‐related quality of life (HRQOL) has received more and more attention as an outcome in cancer therapy. In this exploratory study, we assessed the long‐term HRQOL among 77 surgically treated patients with parotid gland cancer.

Association Between Facial Nerve Monitoring With Postoperative Facial Paralysis in Parotidectomy.

IMPORTANCE Electrophysiologic facial nerve monitoring is becoming an established intraoperative aid to assist the surgeon in facial nerve trunk or branch location and dissection. Limited studies have addressed the postoperative outcomes of parotid surgery with and without monitoring. OBJECTIVE To examine the influence of intraoperative facial nerve monitoring on postoperative facial nerve function and procedure duration in parotid surgery. DESIGN, SETTING, AND PARTICIPANTS An 8-year retrospective review of parotidectomies performed at the Department of Otorhinolaryngology-Head and Neck Surgery, University of Münster. The study analyzed 120 patients undergoing parotidectomy without monitoring from January 1, 1988, to December 31, 1991, and 147 patients undergoing parotidectomy with monitoring from January 1, 2003, to December 31, 2006. The patients were further subdivided in partial parotidectomy (PP) (n = 222) and total parotidectomy (TP) (n = 45) groups. An evaluation of operative time was performed to test the hypothesis of shorter duration of surgery with facial nerve monitoring. Final follow-up was completed on December 31, 2008, and data were analyzed from June 1 to December 31, 2013. MAIN OUTCOMES AND MEASURES Comparison of the incidence of facial nerve dysfunction and operative time between the PP and TP subgroups with and without monitoring. RESULTS A total of 267 patients (127 men [47.6%] and 140 women [52.4%]; mean [SD] age, 51.3 [17.6] years; range, 3-90 years) were included in the analysis. A significant reduction in postoperative facial nerve dysfunction with the use of nerve monitoring could be seen in the PP group (46 of 99 without monitoring [46.5%] vs 18 of 123 with monitoring [14.6%]; P = .001). A similar finding was evident in the TP group when comparing moderate and severe nerve dysfunction (9 of 21 without monitoring [42.9%] vs 2 of 24 with monitoring [8.3%]; P = .01). The mean (SD) operative time in the PP subgroup without nerve monitoring was 115.3 (37.8) minutes; with nerve monitoring, 110.1 (33.6) minutes. The mean (SD) operative time in the TP subgroup without nerve monitoring was 134.5 (50.4) minutes; with nerve monitoring, 158.3 (56.3) minutes. There was no statistical difference between these groups. CONCLUSIONS AND RELEVANCE Facial nerve monitoring in primary parotid surgery for benign and malignant disease does not necessarily reduce the operative time, but the rate of transient postoperative facial nerve dysfunction or the grade of palsy is reduced.

Reliable detection of Human papillomavirus in recurrent laryngeal papillomatosis and associated carcinoma of archival tissue

Recurrent laryngeal papillomatosis (RLP) is, although benign, a challenging disease for both, the patient and the treating physician. Maximum disease control with minimum intervention is considered to be the gold standard. However, patients have to undergo repeating surgical interventions. Human papillomavirus (HPV), mainly so called low risk types, are thought to be responsible for the development of RLP. But, there is still some controversy over the true prevalence of HPV and the virus‐specific molecular diagnostic of choice. Therefore archival tissue samples from 44 patients with RLP at laryngeal site, out of which eight developed laryngeal cancer, was screened for presence of HPV through various molecular approaches. Results from these different methodologies were compared between each other and with patients characteristics. The overall detection rates of HPV with the various methods used in this study were: HPV16 E6/E7 PCR: 0%; GP5+/6+ PCR: 4.5%; CDKN2A/p16 immunohistochemistry: 6.8%; in‐situ hybridization for low and high risk HPV types: 52.3%; HPV6/11 L1 PCR: 72.7% and HPV6/11 E6 PCR: 79.5%. Disease progression showed no apparent dependence of the detected HPV type or clinical variables like age at diagnosis, sex, or additional drug application (Cidofovir and Bevacizumab). In conclusion, the broad‐spectrum PCRs alone or in combination with immunohistochemistry of CDKN2A/p16 and in‐situ hybridization are unsuitable for HPV detection in RLP. Based on the findings presented in this study the type specific PCRs targeting the E6 open reading frame are clearly superior in detection of HPV in this tumor entity. J. Med. Virol. 87:860–870, 2015.

Coexistence of sarcoidosis and metastatic lesions: A diagnostic and therapeutic dilemma (Review)

Sarcoidosis, a chronic, inflammatory disease that affects various different organs, is characterized by noncaseating epitheloid granulomas. This systemic inflammatory process is associated with an increased risk of cancer. Several cases of sarcoidosis that mimic metastatic tumor progression in radiological findings have been reported so far. However, there are also cases that have presented a coexistence of sarcoidosis and metastasis, which have caused a diagnostic and therapeutic dilemma. Due to inadequate current therapies, a reliable differentiation between benign and malignant lesions is crucial. This review focuses on the residual risk of the coexistence of metastases within radiological suspicious lesions in patients with a history of solid tumors and sarcoidosis, as well as immunological findings, in order to explain the potential associations. Sarcoidosis has the potential to promote metastasis as it includes tumor-promoting and immune-regulating cell subsets. Notably, myeloid derived suppressor cells may serve a pivotal role in metastatic progression in patients with sarcoidosis. In addition, the present review also evaluates the potential novel diagnostic approaches, which may be able to differentiate between metastatic lesions and sarcoidosis. The risk of coexistent metastasis in sarcoidosis lesions must be considered by clinical practitioners, and a multidisciplinary approach may be required to avoid misdiagnosis and the subsequent unnecessary surgery or insufficient treatments.