Neck Squamous Cell Carcinoma Cases Research Articles

MAPK Signaling Pathway in Oral Squamous Cell Carcinoma: Biological Function and Targeted Therapy.

Simple SummaryThe aim of the present review was to summarize our studies on the signaling pathways in cancer types that are involved in the development of oral cancer and several mitogen-activated protein kinase-related molecular targeting technologies combined with immune checkpoint therapies to provide new therapeutic strategies for oral squamous cell carcinoma. In addition, we have provided a reasonable outlook, with a systematic basis, for future diagnosis and accurate treatment.Oral squamous cell carcinoma accounts for 95% of human head and neck squamous cell carcinoma cases. It is highly malignant and aggressive, with a poor prognosis and a 5-year survival rate of <50%. In recent years, basic and clinical studies have been performed on the role of the mitogen-activated protein kinase (MAPK) signaling pathway in oral cancer. The MAPK signaling pathway is activated in over 50% of human oral cancer cases. Herein, we review research progress on the MAPK signaling pathway and its potential therapeutic mechanisms and discuss its molecular targeting to explore its potential as a therapeutic strategy for oral squamous cell carcinoma.

Genetic Alterations in HSPA Family of Genes and their Putative Association with HNSCC

Introduction: Head and neck squamous cell carcinoma (HNSCC) is an aggressive life-threatening disease associated with high mortality rates. Several genes related to stress response have been studied extensively to reveal their possible association with HNSCC. Members of the HSPA (heat shock protein family A) family are exclusively expressed under stress conditions suggesting that these are specialised to function in the stress response.&#x0D; Aim: The aim of the present study is to demonstrate the genetic alterations in the HSPA gene family and their putative association with HNSCC.&#x0D; Materials and Methods: The source of the patient's data was procured from the cBioportal database. The TCGA Firehose Legacy comprised 528 head and neck squamous cell carcinoma cases. Oncoprint data analysis can be used to derive a putative association between the disease phenotype and genotype, to identify the variations and to identify any novel variations which can be associated with the disease phenotype. The gnomAD data analysis was used to compare between the observed variants documented in the present study with that of reported variants deposited in the gnomAD repository.&#x0D; Results and Discussion: The HSPA1A&amp;HSPA1B genes harboured high frequency of amplification and deep deletions. The expression of two genes HSPA8 and HSPA13 was found to be up-regulated in the primary tumor sample in comparison to normal tissues. In Kaplan Meier analysis, HSPA8 gene expression was compared with gender, here, high expression in females shows low survival rate. Similarly, HSPA13 gene expression was compared with race, here low expression in africanamerican population shows low survival rate.&#x0D; Conclusion: The present study provides preliminary data on the involvement of the HSPA family of genes with HNSCC, which has to be validated using experimental evidence in other populations.

Correlation of p16 immunostaining in cell-blocks with corresponding tissue specimens for squamous cell carcinomas of the oropharynx.

The goal of this study was to evaluate the performance of p16 staining in cell-blocks vs tissue specimens as a surrogate marker for human papillomavirus (HPV) status in oropharyngeal squamous cell carcinomas. Head and neck squamous cell carcinoma cases presenting as a neck mass with a p16 result on cytology and corresponding tissue specimens (1 January 2014 to 30 June 1920) were included in the study. The following were assessed from cell-block material: number of tumour clusters, percentage of tumour cells with p16 staining, and presence of staining in clusters vs single cells. Results were compared to tissue p16 status. Results of any other ancillary HPV testing were also noted. Forty-two head and neck squamous cell carcinoma neck metastases (35 oropharyngeal, five non-oropharyngeal, and 2 unknown primaries) were identified. The p16 staining pattern in cell-blocks was seen in single cells (27.6%), clusters (44.8%), or both (27.6%). The percentage of tumour cells staining for p16 in cell-blocks was much lower than in corresponding tissue specimens. There were four false negatives and one false positive (concurrent HPV DNA polymerase chain reaction testing was positive in cytology and surgical material). Compared to tissue, the cut-off for p16 interpretation in cell-blocks is substantially lower and staining may be present in single cells or clusters. In 96.9% of cases, any p16 staining in cell-blocks correlated with positive p16 staining in surgical specimens. However, a negative or discrepant p16 result on cell-block should prompt confirmatory HPV studies, as false negative p16 staining in cell-blocks is high.

Abstract 1536: Tumor heterogeneity and primary versus metastatic evaluation of PD-L1

Abstract Immunotherapy with checkpoint inhibitors allowing recovery of effector cell function, has been demonstrated to be highly effective in many tumor types and represents a true revolution in oncology. However, current assays for the prognostic and/or predictive role of tumor PD-L1 expression are not fully standardized with respect to either quantity or distribution of expression, or regarding the concordance of expression in primary tumor versus metastasis. To assess tumor heterogeneity, we evaluated PD-L1 expression using the Ventana SP-263 assay in paired formalin fixed paraffin embedded (FFPE) blocks from the same primary tumor for each of 22 lung squamous cell carcinoma (SCC), 6 lung adenocarcinoma (ADC), 53 gastric ADC, 55 colorectal ADC, 60 urothelial bladder carcinoma, 3 pancreatic ductal ADC and 4 head and neck SCC. In addition, PD-L1 expression in primary tumor and synchronous metastasis was evaluated in 32 cases of lung neoplasm (10 ADC +22 SCC), 30 cases of gastric adenocarcinoma, 55 cases of colorectal adenocarcinoma, and 16 head and neck SCC, each with corresponding synchronous metastases. PD-L1 expression was determined by pathologist evaluation of the percentage of positive tumor cells and cases were grouped into categories representing &amp;lt;1%, 2-9%, 10-25%, 26-49%, 50-75% and 76-100%, with those samples expressing more than 25% tumor positivity considered as expressing “high” PD-L1 levels. In lung carcinoma, just 1 case of SCC exhibited a change in PD-L1 classification when two blocks from the same tumor were evaluated (0% vs 5%), and also for 1 case of ADC (2% vs 0%) . In gastric ADC we recorded a change in classification in 1 out of 53 cases (5% vs 0%), and for CRC ADC we found a difference between the two blocks evaluated in just 1 of the 55 cases (0% vs 50%). For the remaining tumor cohorts (60 bladder carcinoma, 4 head and neck SCC and 3 pancreatic ADC) 100% concordance was observed between two blocks from the same tumor. Evaluation of primary tumors and synchronous metastasis revealed no differences in PD-L1 scores for the 32 cases of lung carcinoma, or for the 16 head and neck SCC cases. In the 30 gastric ADC cases, there was a discordant PD-L1 expression score for just one pair of tumors (primary tumor 3% and metastasis 0%). Similarly, 1 of 55 CRC ADC examined revealed a primary tumor PD-L1 score of 50% while the metastatic tissue was negative. Taken together, these data demonstrate that PD-L1 expression between two sites in the same tumour, and between primary versus synchronous metastases is remarkably consistent. The conclusions drawn are that a representative PD-L1 score may be achieved from a single FFPE tumour block, and that for cases where obtaining tissue from a primary tumor may be challenging, PD-L1 could be evaluated in metastatic tissue. Citation Format: Lorenzo Colarossi, Eleonora Aiello, Marie Cumberbatch, Cristina Colarossi, Chris Womack, Lorenzo Memeo. Tumor heterogeneity and primary versus metastatic evaluation of PD-L1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1536.

Cell-free mitochondrial DNA copy number variation in head and neck squamous cell carcinoma: A study of non-invasive biomarker from Northeast India.

Head and neck squamous cell carcinoma is the most commonly diagnosed cancer worldwide. The lifestyle, food habits, and customary practices manifest the Northeast Indian population toward higher susceptibility to develop head and neck squamous cell carcinoma. Here, we have investigated the association of smoke and smokeless tobacco, and alcohol with copy number variation of cell-free mitochondrial DNA and cell-free nuclear DNA in cases and controls. Cell-free DNA from plasma was isolated from 50 head and neck squamous cell carcinoma cases and 50 controls with informed written consent using QIAamp Circulating Nucleic Acid Kit. Real-time polymerase chain reaction was done for copy number variation in cell-free mitochondrial DNA and cell-free nuclear DNA. Receiver operating characteristic curve analysis was performed to evaluate the diagnostic application between the two study groups using clinicopathological parameters. The levels of cell-free nuclear DNA and cell-free mitochondrial DNA of cases in association with smoke and smokeless tobacco, alcohol with smoking (p < 0.05) were significantly higher (p < 0.01 and p < 0.001, respectively) than controls. Using receiver operating characteristic curve analysis between head and neck squamous cell carcinoma cases and controls, we distinguished cell-free mitochondrial DNA (cutoff: 19.84 raw Ct; sensitivity: 84%; specificity: 100%; p < 0.001) and cell-free nuclear DNA (cutoff: 463,282 genomic equivalent/mL; sensitivity: 53%; specificity: 87%; p < 0.001). The copy number variation in cases (cell-free nuclear DNA: 5451.66 genomic equivalent/mL and cell-free mitochondrial DNA: 29,103,476.15 genomic equivalent/mL) and controls (cell-free nuclear DNA: 1650.9 genomic equivalent/mL and cell-free mitochondrial DNA: 9,189,312.54 genomic equivalent/mL), respectively. Our result indicates that the cell-free mitochondrial DNA content is highly associated with smoke and smokeless tobacco, betel quid chewing, and alcohol which shows greater promises, holding the key characteristics of diagnostic biomarkers, that is, minimal invasiveness, high specificity, and sensitivity.

Immunohistochemical and molecular imaging biomarker signature for the prediction of failure site after chemoradiation for head and neck squamous cell carcinoma

Objective: To identify a failure site-specific prognostic model by combining immunohistochemistry (IHC) and molecular imaging information to predict long-term failure type in squamous cell carcinoma of the head and neck.Patient and methods: Tissue microarray blocks of 196 head and neck squamous cell carcinoma cases were stained for a panel of biomarkers using IHC. Gross tumor volume (GTV) from the PET/CT radiation treatment planning CT scan, maximal Standard Uptake Value (SUVmax) of fludeoxyglucose (FDG) and clinical information were included in the model building using Cox proportional hazards models, stratified for p16 status in oropharyngeal carcinomas. Separate models were built for time to locoregional failure and time to distant metastasis.Results: Higher than median p53 expression on IHC tended toward a risk factor for locoregional failure but was protective for distant metastasis, χ2 for difference p = .003. The final model for locoregional failure included p53 (HR: 1.9; p: .055), concomitant cisplatin (HR: 0.41; p: .008), β-tubulin-1 (HR: 1.8; p: .08), β-tubulin-2 (HR: 0.49; p: .057) and SUVmax (HR: 2.1; p: .046). The final model for distant metastasis included p53 (HR: 0.23; p: .025), Bcl-2 (HR: 2.6; p: .08), SUVmax (HR: 3.5; p: .095) and GTV (HR: 1.7; p: .063).Conclusions: The models successfully distinguished between risk of locoregional failure and risk of distant metastasis, which is important information for clinical decision-making. High p53 expression has opposite prognostic effects for the two endpoints; increasing risk of locoregional failure, but decreasing the risk of metastatic failure, but external validation of this finding is needed.

RNA in-situ hybridization is a practical and effective method for determining HPV status of oropharyngeal squamous cell carcinoma including discordant cases that are p16 positive by immunohistochemistry but HPV negative by DNA in-situ hybridization

Evaluation of human papillomavirus (HPV) status in oropharyngeal squamous cell carcinoma (OPSCC) has become increasingly important for prognostication and clinical trial enrollment. This assessment is confounded in OPSCCs that are p16 positive by immunohistochemistry (IHC) but HPV negative by DNA in situ hybridization (DISH). This study evaluates whether E6/E7 mRNA in situ hybridization (RISH) can detect transcriptionally active HPV in these problematic cases. Eighty-two head and neck squamous cell carcinoma cases that had previously undergone p16 IHC and HPV DISH were evaluated with two RISH platforms and a second-generation DISH probe. The study included 21 p16+/DISH+ concordant cases, 19 p16-/DISH- concordant cases, and 42 p16+/DISH- discordant cases. RISH identified E6/E7 mRNA in 37 (88%) p16+/DISH- cases, 21 (100%) p16+/DISH+ cases, and 0 (0%) p16-/DISH- cases. RISH signals were clearly visible at low to medium magnification in 97% of positive cases, facilitating almost-perfect inter-observer reproducibility. The performance of the manual and automated RISH platforms were equivalent (kappa=0.915). Only 29% of carcinomas that demonstrated E6/E7 mRNA transcriptional activity were positive using the 2nd generation DISH probe. HPV RISH is a highly sensitive and specific platform that can clarify the HPV status of those perplexing OPSCCs that are p16 positive by IHC but HPV negative by DISH. Moreover, it is easy to interpret, readily adaptable to the clinical laboratory, and provides direct evidence of HPV transcriptional activity. E6/E7 RISH should be considered as a first-line platform for determination of HPV status in OPSCCs.

Alteration of serum lipid profile and its prognostic value in head and neck squamous cell carcinoma.

Several serum lipid components have been implicated in the development of cancer. However, the prognostic significance of serum lipid components in head and neck squamous cell carcinoma is unknown. Here, we investigated the predictive value of serum lipid profile at diagnosis and in the overall survival of the patients. The study population consists of 136 pathologically confirmed head and neck squamous cell carcinoma cases diagnosed between years 2009 and 2014 at a tertiary medical center. Levels of preoperative serum lipid component's total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein, apolipoprotein A, apolipoprotein B, and lipoprotein (a) were compared between patients and normal controls matched for age and gender. Serum lipid profiles and their association with clinical parameters were analyzed. The effects of the serum lipid components on survival were examined using the proportional hazards regression model to estimate hazard ratio. Significant lower levels of cholesterol, low-density lipoprotein, apolipoprotein A, and apolipoprotein B were found in patients with oral cancer (P < 0.0001). However, a significantly higher level of lipoprotein (a) was found in the cancer group (P < 0.0001). Patients with higher lipoprotein (a) had significantly shorter overall survival than those with lower lipoprotein (a) (P = 0.0042). Multivariate analysis showed that both higher lipoprotein (a) and lymph node metastasis are independent prognostic factors in the patient population (P < 0.01). A higher lipoprotein (a) was associated with poorer prognosis and might be a novel marker in patients with head and neck squamous cell carcinoma.

Sarcomatoid Carcinoma in Head and Neck

Sarcomatoid carcinoma (SaCa) is a rare variant of squamous cell carcinoma (SCC) with sarcomatoid features. This study investigated the clinical presentation and outcomes of head and neck SaCa. In addition, reconstructive outcome for a subset of patients was also evaluated. Seventy-eight SaCa cases including 72 men and 6 women were identified from 13,777 head and neck SCC cases. Clinical outcomes were evaluated based on locoregional control, distant metastases, and multivariate analyses. Reconstructive outcome was evaluated by flap survival rate. Of the 78 cases, 71% (55) of cases were located in the oral mucosa; 64% (50) of patients were classified as T3 or T4 at the time of diagnosis. The 5-year survival was only 16%. Multivariate analysis revealed better outcomes only when the patient had a history of previous SCC. Forty-five patients underwent flap reconstruction, with 98% flap survival rate but the functional result varied because of the inevitable adjuvant radiotherapy and advanced stage of tumor. Sarcomatoid carcinoma is a different entity from the conventional SCC of the head and neck. Sarcomatoid carcinoma carries a poorer prognosis despite aggressive surgical intervention and concurrent adjuvant therapies. It remains a great challenge for clinical oncologists, and the optimal treatment strategy requires further studies. Free flap is still preferred for defect reconstruction but the design should be simplified to avoid complications.

Value of Postoperative Radiation Therapy for Regional Control after Dissection in Head and Neck Squamous Cell Carcinoma Cases

We aimed to define clinicopathologic risk factors associated with regional recurrence (RR) and thus the effectiveness of postoperative radiotherapy (PORT) for neck control for head and neck squamous cell carcinomas (HNSCCs) with differing cervical lymph node status. A retrospective study was performed in 196 HNSCC patients with pathologically positive neck node (N+) to evaluate the high-risk factors for RR and to define the role of PORT in control after neck dissection and postoperative radiotherapy (PORT). Overall, the RR rate after neck dissection and PORT was 29%. Extracapsular spread (ECS) was confirmed to be the only independent risk factor for RR. There were no significant risk factors associated with RR in the ECS- group. The 5-year disease-specific survival rate was 45%, which descended to 10% with the emergence of RR. ECS remains a determined risk factor for RR after neck dissection and PORT in patients with N+. PORT alone is not adequate for preventing RR in the neck with ECS after neck dissection. More intensive postoperative adjuvant therapies, especially combined chemotherapy and radiotherapy, are needed to prevent regional failure in HNSCC patients with ECS.

Immunotherapy for Head and Neck Cancer Patients: Shifting the Balance

Head and neck squamous cell carcinoma is the sixth most common cancer in the western world. Over the last few decades little improvement has been made to increase the relatively low 5-year survival rate. This calls for novel and improved therapies. Here, we describe opportunities in immunotherapy for head and neck cancer patients and hurdles yet to be overcome. Viruses are involved in a subset of head and neck squamous cell carcinoma cases. The incidence of HPV-related head and neck cancer is increasing and is a distinctly different disease from other head and neck carcinomas. Virus-induced tumors express viral antigens that are good targets for immunotherapeutic treatment options. The type of immunotherapeutic treatment, either active or passive, should be selected depending on the HPV status of the tumor and the immune status of the patient.

Highlights of This Issue

Weight change after a breast cancer diagnosis has been linked to lower survival. To investigate this relationship, Caan and colleagues analyzed postdiagnostic weight variation and survival in breast cancer patients from the United States and China. They report that weight stability in the early years following diagnosis was associated with lower overall mortality risk. In the United States, lower survival was seen for women who lost weight after breast cancer diagnosis and had comorbid conditions. This report illuminates the importance of personalized weight control strategies for breast cancer survivors.The field of cancer epidemiology has been criticized for producing false-positive associations. Demetriou and colleagues investigated the frequency of and factors contributing to false-positive findings in cancer epidemiology studies. The International Agency for Research on Cancer Monographs Group 3 agents were examined to identify potential false-positive findings, and frequency estimates for false-positive occurrences were calculated. Among 509 agents, only 37 were found to have potential false-positive associations. These results suggest that cancer epidemiology is not overwhelmed with false-positive findings. This study should help restore faith in epidemiologic procedures and validate epidemiologic findings as important tools to guide public health measures.Studies on the use of fertility drugs and ovarian cancer risk have produced conflicting results. Kurta and colleagues used data from the Hormones and Ovarian Cancer Prediction (HOPE) study to determine if fertility drug use has an impact on ovarian cancer risk, taking into account parity, gravidity, and cause of infertility. The authors report that ever use of fertility drugs was not significantly associated with ovarian cancer within the total HOPE population or among women who sought attention for infertility. These results suggest that fertility drug use does not significantly contribute to overall risk of ovarian cancer when adjustment is made for known confounding factors.Blood leukocytes from patients with solid tumors exhibit complex and distinct cancer-associated patterns of DNA methylation. Koestler and colleagues examined this by first identifying differentially methylated regions (DMR) in leukocytes from healthy control subjects. They then evaluated these leukocyte DMRs in patients with different cancers. They report that a substantial proportion of the most common leukocyte DMRs were differentially methylated to a statistically significant extent among head and neck squamous cell carcinoma cases and ovarian cancer cases, compared with cancer-free control subjects. These results suggest that leukocyte DMRs could represent powerful new diagnostic tools, applicable to a range of solid tumors.

Analysis of Microsatellite Instability in Laryngeal Squamous Cell Carcinoma

Introduction and objectivesThe literature on the involvement of microsatellite instability in head and neck squamous cell carcinoma shows great variability, probably due to differences in the testing methods. Using a consensus detection system, we aimed to reach a reliable estimate of microsatellite instability prevalence in a subset of head and neck squamous cell carcinoma cases. MethodsThe microsatellite instability status of 43 patients with previously untreated primary laryngeal squamous cell carcinomas was analysed by a multiplex polymerase chain reaction assay including 5 mononucleotide repeat markers. ResultsThirty-six cases showed a stable phenotype or a microsatellite stable phenotype (83.7%) and 7 cases (16.3%) showed a microsatellite instability-positive phenotype. One case showed instability in 3 of 5 markers, 1 case in 2 markers and 5 cases in 1 marker. The microsatellite instability-positive and stable cases did not differ with respect to age, tumour stage, lymph node or distant metastases. ConclusionsOur data showed that a proportion of laryngeal squamous cell carcinomas are microsatellite instability positive. Knowledge of microsatellite instability patient status will allow adjusting anticancer therapy at an individual level.

Análisis de la inestabilidad de microsatélites en el carcinoma escamoso de laringe

Introduction and objectivesThe literature on the involvement of microsatellite instability in head and neck squamous cell carcinoma shows great variability, probably due to differences in the testing methods. Using a consensus detection system, we aimed to reach a reliable estimate of microsatellite instability prevalence in a subset of head and neck squamous cell carcinoma cases. MethodsThe microsatellite instabilityI status of 43 patients with previously untreated primary laryngeal squamous cell carcinomas was analyzed by a multiplex polymerase chain reaction assay including 5 mononucleotide repeat markers. ResultsThirty-six cases showed a stable phenotype or a microsatellite stable phenotype (83.7%) and 7 cases (16.3%) showed an microsatellite instability-positive phenotype. One case showed instability in 3 of 5 markers, 1 case in 2 markers and 5 cases in 1 marker. The microsatellite instability-positive and stable cases did not differ with respect to age, tumour stage, lymph node or distant metastases. ConclusionsOur data showed that a proportion of laryngeal squamous cell carcinomas are microsatellite instability positive. Knowledge of microsatellite instability patient status will allow adjusting anticancer therapy at an individual level.

Gene Expression Profiling Identifies Genes Predictive of Oral Squamous Cell Carcinoma

Oral squamous cell carcinoma (OSCC) is associated with substantial mortality and morbidity. To identify potential biomarkers for the early detection of invasive OSCC, we compared the gene expressions of incident primary OSCC, oral dysplasia, and clinically normal oral tissue from surgical patients without head and neck cancer or preneoplastic oral lesions (controls), using Affymetrix U133 2.0 Plus arrays. We identified 131 differentially expressed probe sets using a training set of 119 OSCC patients and 35 controls. Forward and stepwise logistic regression analyses identified 10 successive combinations of genes which expression differentiated OSCC from controls. The best model included LAMC2, encoding laminin-gamma2 chain, and COL4A1, encoding collagen, type IV alpha1 chain. Subsequent modeling without these two markers showed that COL1A1, encoding collagen, type I alpha1 chain, and PADI1, encoding peptidyl arginine deiminase, type 1, could also distinguish OSCC from controls. We validated these two models using an internal independent testing set of 48 invasive OSCC and 10 controls and an external testing set of 42 head and neck squamous cell carcinoma cases and 14 controls (GEO GSE6791), with sensitivity and specificity above 95%. These two models were also able to distinguish dysplasia (n = 17) from control (n = 35) tissue. Differential expression of these four genes was confirmed by quantitative reverse transcription-PCR. If confirmed in larger studies, the proposed models may hold promise for monitoring local recurrence at surgical margins and the development of second primary oral cancer in patients with OSCC.