Diagnosis Of HNSCC Research Articles

Extracellular Vesicle microRNAs as Possible Liquid Biopsy Markers in HNSCC-A Longitudinal, Monocentric Study.

Biomarkers for HNSCC are still lacking. Biomolecules obtained via liquid biopsy are being investigated for diagnosis, prognosis, and therapy monitoring, including extracellular vesicles (EVs) and EV-cargo, e.g., proteins, RNA, and microRNA. This study aims to understand localization-dependent EV-microRNA expression in blood sera, their dynamics over time (12 months FU), and insights into their potential in diagnostics and therapy monitoring. Via liquid biopsy, blood serum was taken from 50 patients with HNSCC and 16 controls. Extracellular vesicles were isolated from serum by precipitation, and the contained microRNA-21, -1246, -200c, -let-7a, -181a, and -26a were amplified by reverse transcription and determined with real-time PCR. Expression ratios (HNSCC to healthy controls) were collated with the patients' clinical parameters. A second liquid biopsy was carried out avg. 12 months later in the tumor aftercare. A sub-analysis with the Oropharynx subsite was implemented. EV-mir-21, -let-7a, and -181a were 2.5-3-fold higher expressed in HPV/p16+ than in HPV/p16- HNSCC. Different expressions of EV-mir-181a and -26a could be demonstrated depending on the therapy modality. EV-microRNA could be a promising biomarker in the diagnosis and therapy monitoring of HNSCC. A systematic comparison of EV- and tissue microRNA expression in different HNSCC-subsites is needed.

Assessing the use of the novel tool Claude 3 in comparison to ChatGPT 4.0 as an artificial intelligence tool in the diagnosis and therapy of primary head and neck cancer cases

ObjectivesHead and neck squamous cell carcinoma (HNSCC) is a complex malignancy that requires a multidisciplinary tumor board approach for individual treatment planning. In recent years, artificial intelligence tools have emerged to assist healthcare professionals in making informed treatment decisions. This study investigates the application of the newly published LLM Claude 3 Opus compared to the currently most advanced LLM ChatGPT 4.0 for the diagnosis and therapy planning of primary HNSCC. The results were compared to that of a conventional multidisciplinary tumor board; (2) Materials and Methods: We conducted a study in March 2024 on 50 consecutive primary head and neck cancer cases. The diagnostics and MDT recommendations were compared to the Claude 3 Opus and ChatGPT 4.0 recommendations for each patient and rated by two independent reviewers for the following parameters: clinical recommendation, explanation, and summarization in addition to the Artificial Intelligence Performance Instrument (AIPI); (3) Results: In this study, Claude 3 achieved better scores for the diagnostic workup of patients than ChatGPT 4.0 and provided treatment recommendations involving surgery, chemotherapy, and radiation therapy. In terms of clinical recommendations, explanation and summarization Claude 3 scored similar to ChatGPT 4.0, listing treatment recommendations which were congruent with the MDT, but failed to cite the source of the information; (4) Conclusion: This study is the first analysis of Claude 3 for primary head and neck cancer cases and demonstrates a superior performance in the diagnosis of HNSCC than ChatGPT 4.0 and similar results for therapy recommendations. This marks the advent of a newly launched advanced AI model that may be superior to ChatGPT 4.0 for the assessment of primary head and neck cancer cases and may assist in the clinical diagnostic and MDT setting.

CCDC71L as a novel prognostic marker and immunotherapy target via lipid metabolism in head and neck squamous cell carcinoma

BackgroundHead and neck squamous cell carcinoma (HNSCC) is the sixth most widespread cancer globally with high rate and poor prognosis. Coiled-coil domain containing 71 like (CCDC71L) exerts an important role in cellular lipid metabolic process. However, its function in HNSCC remains unclear. To this end, we examined the CCDC71L implications for prognosis and tumor microenvironment in HNSCC. Materials and methodsFirst, CCDC71L expression was explored through The Cancer Genome Atlas (TCGA), Human Protein Atlas (HPA), the Gene Expression Profiling Interactive Analysis 2 (GEPIA2), Clinical Proteomic Tumor Analysis Consortium (CPTAC) and the Gene Expression Omnibus (GEO) databases. The clinicpathological information were obtained from the dataset of TCGA. The Kaplan–Meier Plotter databases and Cox model were performed for the determination of prognostic values of CCDC71L, including the overall survival (OS), progress free interval (PFI), recurrence-free survival (RFS) and disease specific survival (DSS). Then, the potential mechanism of CCDC71L in HNSCC development was elucidated by means of Gene set enrichment analysis (GSEA) and Metascape databases. Furthermore, the relevance of CCDC71L to immune cells infiltration and immune checkpoints was assessed. The correlations among CCDC71L expression, mutational landscape and genome heterogeneity [mutant-allele tumor heterogeneity (MATH) and tumor purity] were detected by the data in TCGA. ResultsCCDC71L expression was significantly upregulated in HNSCC, and positively associated with age, gender and N stage. Higher CCDC71L expression resulted in poor OS, RFS, DSS and PFI. Multivariate Cox regression analysis showed CCDC71L would be an independent prognostic marker in patients with HNSCC. Moreover, CCDC71L and the level of macrophage and neutrophil cells infiltration were significantly correlated in HNSCC. High expression of CCDC71L was related to immune checkpoint genes, oncogene mutations and genome heterogeneity markers. ConclusionThese results implied that CCDC71L plays vital roles in HNSCC progression, which could be used as a underlying biomarker for the diagnosis and prognosis of HNSCC. Meanwhile, CCDC71L participates in immune regulation, which has a potential value for the immunotherapy of HNSCC patients.

Abstract 6527: Fusion genes as novel putative biomarkers for head and neck squamous cell carcinomas

Abstract Head and neck squamous cell carcinomas (HNSCC) are highly aggressive types of cancers with a complex molecular etiology of disease and consequently result in delayed clinical detection, poor prognosis, and limited treatment strategies. Fusion genes, a hallmark of cancer, may be targeted for early identification and treatment of HNSCC; however, studies characterizing fusion genes in HNSCC remain limited. Oral Human Papillomavirus (HPV) infection is a major cause of HNSCC and can also promote the acquisition of fusion genes due to the genomic instability induced by its integration. Here, we retrieved RNA-sequencing data from 36 HNSCC tumors (18 HPV+ and 18 HPV-, GEO Accession: GSE74956) to identify fusion genes using the STAR fusion algorithm in each sample. Statistical analyses were performed in R statistical software. A total of 10 fusion genes were identified in at least 4 of the samples regardless of HPV status. We identified 3 fusion genes that were present in at least 4 of the HPV+ tumors: PMS2P9-CCDC146, MACC1-AC005062.1, AC096711.2-RPL7AP28, and one solely present in HPV- tumors: CTSC-RAB38. The remaining 6 fusion genes were present in both HPV+ and HPV- tumors: TVP23C-CDRT4, AC009271.1-LINC01905, MIR205HG-AOPEP, SEPTIN7P2-PSPH, KRT5-KRT84, KANSL1-ARL17A. While 4 of the identified fusion genes had been previously associated with cancer, there were 6 novel identified fusion genes, in which one or both partner genes have been associated with different cancers. Additionally, the identified fusion genes may be a product of transcription read-through of neighbor genes. The fusion genes associated with HPV infection might explain biological disparities in HPV+ versus HPV- HNSCC and may be targets for the prevention and diagnosis of HNSCC. Future studies need to determine the functional impact of the identified fusion genes in HPV+ tumors. Citation Format: Yabdiel A. Ramos Valerio, Hannah C. Van Wyk, Esther Peterson Peguero, Josué Pérez Santiago. Fusion genes as novel putative biomarkers for head and neck squamous cell carcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6527.

Progress of Tumor-Derived Exosomes in Head and Neck Squamous Cell Carcinoma

Exosomes are extracellular vesicles 40 to 160 nm in diameter that mainly mediate information transmission and substance exchange between cells, and increasing evidence suggests that exosomes are involved in the development of tumors and have great potential in cancer diagnosis and treatment. Squamous cell carcinoma of head and neck is one of the common malignant tumors in the whole body. Although various treatments continue to develop, the 5-year survival rate remains only 50%, which may be related to its characteristics such as easy lymphatic metastasis, chemotherapy resistance and poor radiotherapy sensitivity. Researchers are paying more and more attention to TDE, and studies connected to it have significantly improved our understanding of the causes, development, diagnosis, and prognosis of HNSCC. Head and neck squamous cell carcinoma-derived exosomes involve tumor initiation, progression, immune regulation, diagnosis and treatment application, and are anticipated to serve as therapeutic targets and biomarkers for early tumor diagnosis, and new ideas for improving the survival rate and prognosis of patients with head and neck squamous cell carcinoma are provided by their related studies. Here, in order to bring our knowledge of exosomes derived from head and neck squamous cell carcinoma up to date, we examine the development of research on tumor-derived exosomes in the disease.

Addictions, Social Deprivation and Cessation Failure in Head and Neck Squamous Cell Carcinoma Survivors

To evaluate the evolution of addictions (tobacco and alcohol) and social precarity in head and neck squamous cell carcinoma survivors when these factors are addressed from the time of diagnosis. Addictions and social precarity in patients with a new diagnosis of HNSCC were assessed through the EPICES score, the Fagerström score, and the CAGE questionnaire. When identified as precarious/dependent, patients were referred to relevant addiction/social services. One hundred and eighty-two patients were included. At the time of diagnosis, an active tobacco consumption was associated with alcohol drinking (Fisher's exact test, p < 0.001). Active smokers were more socially deprived (mean EPICES score = mES = 36.2 [±22.1]) than former smokers (mES = 22.8 [±17.8]) and never smokers (mES = 18.9 [±14.5]; Kruskal-Wallis, p < 0.001). The EPICES score was correlated to the Fagerström score (Kruskal-Wallis, p < 0.001). Active drinkers (mES = 34.1 [±21.9]) and former drinkers (mES = 32.7 [±21]) were more likely to be socially deprived than those who never drank (mES = 20.8 [±17.1]; Krukal-Wallis, p < 0.001). A Fagerström score improvement at one year was associated to a CAGE score improvement (Fisher's exact test, p < 0.001). Tobacco and alcohol consumption were more than halved one year after treatment. Patients who continued to smoke one year after diagnosis were significantly more likely to continue to drink (Fisher's exact test, p < 0.001) and had a significantly higher initial EPICES score (Kruskal-Wallis, p < 0.001). At one year, addictions and social deprivation tend to improve when taken care of from the diagnosis. The most dependent patients and those with multiple frailties are at highest risk of cessation failure.

Identification of novel hub genes associated with lymph node metastasis of head and neck squamous cell carcinoma by completive bioinformatics analysis.

BackgroundHead and neck squamous cell carcinoma (HNSCC) is one of the most serious diseases affecting populations worldwide and lymph node metastasis is a key pathological feature of HNSCC which predicts poor survival. However, the molecular mechanisms associated with the development of lymph node metastasis in HNSCC have not been fully elucidated.MethodsDifferentially expressed genes (DEGs) were identified in two HNSCC datasets (GES6631 and GES58911). Functional annotation analysis was constructed via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Meanwhile, the protein-protein interaction (PPI) network and module analysis using Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) and Cytoscape were carried out to identify the hub genes. The expression differences, overall survival (OS), and disease-free survival (DFS) of hub genes were analyzed by Gene Expression Profiling Interactive Analysis 2 (GEPIA2) and verified by immunohistochemistry (IHC) from Human Protein Atlas (HPA). Moreover, receiver operating characteristic (ROC) curve analysis was conducted to judge whether those hub genes had good diagnostic and prognostic ability, and the web tool Tumor Immune Estimation Resource (TIMER) was used to analyze the correlation of hub genes’ expression and immune infiltration.ResultsIn total, 913 DEGs including 476 upregulated and 437 downregulated genes were identified. The genes Aurora kinase A (AURKA), CyclinB1 (CCNB1), Cyclin-dependent kinase regulatory subunit 1B (CKS1B), Serpin Family H Member 1(SERPINH1), and Transforming growth factor-beta-induced protein (TGFBI) were screened out as hub genes and were associated with lymph node metastasis, showing notably abnormal expression in HNSCC samples, and the high expression of all the hub genes in HNSCC patients was related to worse overall survival.ConclusionsThe genes AURKA, CCNB1, CKS1B, SERPINH1, and TGFBI may be involved in the lymph node metastasis of HNSCC and reveal the potential to serve as molecular biomarkers in the diagnosis of HNSCC. This study may help to elucidate the molecular mechanisms of the development of lymph node metastasis and facilitate the selection of targets for the treatment and diagnosis of HNSCC.

Development and Validation of an Autophagy-Related Signature for Head and Neck Squamous Cell Carcinoma.

Introduction HNSCC is the sixth most frequent type of malignant carcinoma with a low prognosis rate. In addition, autophagy is important in cancer development and progression. The purpose of this study is to investigate the potential significance of ARGs in the diagnosis and treatment of HNSCC. Materials and Methods Expression data and clinical information of HNSCC samples were collected from the TCGA database, and a list of ARGs was obtained from the MSigDB. Then, we used R software to perform differential expression analysis and functional enrichment analysis. Further analysis was also performed to find out the survival-related ARGs in HNSCC, and two prognosis-related ARGs, FADD and NKX2-3, were selected to construct a prognosis prediction model. Moreover, some methods were applied to validate the prognosis prediction model. Finally, we used cell lines and clinical tissue samples of HNSCC to analyze the importance of FADD and NKX2-3. Results We screened a total of 38 differentially expressed ARGs, and enrichment analysis showed that these genes were mainly involved in autophagy. Then, we selected FADD and NKX2-3 to construct a prognosis model and the risk score calculated by the model was proved to be effective in predicting the survival of HNSCC patients. Additionally, significant differences of the clinicopathological parameters could also be observed in the risk scores and the expression of NKX2-3 and FADD. The expression of FADD and NKX2-3 in cell lines and HNSCC tissue samples also showed the same trends. Conclusions ARGs may be a potential biomarker for HNSCC prognosis, and targeted therapies for FADD and NKX2-3 are possible to be a new strategy of HNSCC treatment.

The impact of smoking habit alteration on prognosis of head and neck cancer patients

The influence of smoking on survival in patients with HNSCC is well documented in the literature. There is little data on changes in smoking habits after diagnosis. Here, the effect on survival of the reduction of smoking compared to full smoking cessation is investigated. Patient records and tumor documentation of 643 consecutive HNSCC cases of the Head and Neck Tumor Center of the University Hospital Kiel are evaluated retrospectively: smoking habits before and after treatment and survival are evaluated. Change in smoking behavior at the initial diagnosis of HNSCC leads to a significant positive effect on the prognosis compared to continued smoking. There is no difference between smoke reduction and weaning. This effect is based solely on those patients who are treated exclusively by surgery. Lifelong non-smokers have a significant survival advantage over active and ex-smokers, with no difference between the latter two groups. The positive influence of changed smoking habits on the prognosis runs parallel to the negative direct effect of active smoking on therapy, which is attributed to peritumoral hypoxia with a negative effect on the effectiveness of R(C)T. The positive effect of the change in smoking behaviour during surgery alone is most likely due to reduced peri-operative complications. Patients should be encouraged to at least minimize smoking with the cancer diagnosis. In addition, former smokers should be considered active smokers for survival estimates and therapy planning.

MicroRNA as a Novel Biomarker in the Diagnosis of Head and Neck Cancer.

Head and neck squamous cell carcinoma is the sixth most common cancer worldwide, with 890,000 new cases and 450,000 deaths in 2018, and although the survival statistics for some patient groups are improving, there is still an urgent need to find a fast and reliable biomarker that allows early diagnosis. This niche can be filled by microRNA, small single-stranded non-coding RNA molecules, which are expressed in response to specific events in the body. This article presents the potential use of microRNAs in the diagnosis of HNSCC, compares the advances in this field to other diseases, especially other cancers, and discusses the detailed use of miRNA as a biomarker in profiling and predicting the treatment outcome with radiotherapy and immunotherapy. Potential problems and difficulties related to the development of this promising technology, and areas on which future research should be focused in order to overcome these difficulties, were also indicated.

Droplet digital PCR of tumor suppressor gene methylation in serial oral rinses of patients with head and neck squamous cell carcinoma

BackgroundHead and neck squamous cell carcinoma (HNSCC) currently lacks sensitive approaches to detect cancer‐related traits in body fluid.MethodsMethylation of tumor suppressor genes (TSGs) (PAX5, EDNRB, and DCC) were measured in the oral rinses from 50 HNSCC and 58 control subjects using droplet digital PCR (ddPCR). Diagnostic accuracies in detecting HNSCC and the detection rate of recurrence in the post‐treatment monitoring were analyzed.ResultsddPCR TSG methylation detection in oral rinses for diagnosis of HNSCC had an AUC of 0.892 for PAX5, 0.753 for EDNRB, and 0.729 for DCC. Significant drop of TSG methylation was observed after completion of surgery (p < 0.01). 76.9% of the relapse cases had a pre‐emptive rebound of methylation above presurgery levels in at least one of the tested markers before confirmed recurrence.ConclusionsUtilizing ddPCR for TSG methylation detection in oral rinses shows potential for detection and monitoring of HNSCC.

Abstract PO-089: Profiling of salivary proteome specific for the diagnosis of head and neck cancer among African Americans

Abstract Head and Neck Squamous Cell Carcinoma (HNSCC) is one the types of Cancer in The United States with inequality in the mortality rate among Black patients when compared to White patients. Patients of European dissent (Whites) are more likely to be diagnosed with any type of HNSCC but patients of African dissents (Blacks) are more likely to die from ant type of HNSCC when all patients are HPV negative. The difference in survival based on race and ethnicity prompted the need for a better and early diagnostic marker for HNSCC among Blacks and other groups with poor prognosis of HNSCC. The present research seeks to identify a specific and safe salivary proteome marker for the early diagnosis of HNSCC. The hypothesis of this article is that there is a significant difference in the salivary proteome of AA patients with HNSCC when compared to WA patients which is specific to cancer site. Eight patient samples with confirmed diagnosis of Laryngeal cancer at Nashville General hospital (IRB Protocol Number 14-03-172) were collected with appropriate informed consent. Patient samples were pooled together and analyzed using IHC for the presence of shared protein expression. Each protein sample was later analyzed individually using IHC in order to obtain specific proteins expressed by each patient’s salivary proteome. A Multidimensional Protein Identification Technology (MUDPIT) was used to analyze salivary samples to increase resolution for identifying proteins and peptides. MUDPIT was used to increase resolution of analyzed proteins and was compared with other proteins associated with LaCa using a known online protein database called WebGestalt. The most highly expressed proteins in each of the samples were further analyzed using The Human Protein Atlas in order to study the location of the salivary proteome and its function when expressed. 20 unique protein genes obtained from Label Free Quantification (LFQ) analysis was further analyzed for the function of the genes and gene co-localization in the body using an online database known as GeneMANIA. Finally, PEAKS was used to ascertain the effect of proteins. Result shows differences in the salivary proteome of AA patients compared to WA’s. Specifically, proteins involved in transcriptional mis-regulation were over- represented in AA&amp;gt;WA patients which included some proteins associated with metastasis (JUP), dis-functional immune cells (CD14), over-expressed tumor- suppressor proteins (DMBT1 and DRB2) and elevated level of antimicrobial innate immunity (His 1 &amp; 3) are all higher than the normal values in healthy patients without LaCa both among AA and WA. Future research is required to expand on these identified proteins to help provide a safe and specific biomarker for the diagnosis of LaCa among AA patients. Future research can also provide direction as to where therapeutics can be directed and how they can be safely administered to generate the most effective treatment that can improve the prognosis of the disease among AA patients and others with groups with poor prognosis. Citation Format: Oyinloye A. Jose, Oladipupo Anibire, Derek Wagner, Dana Marshall, Billy Ballard, Siddharth Pratap, Victor Paramov. Profiling of salivary proteome specific for the diagnosis of head and neck cancer among African Americans [abstract]. In: Proceedings of the AACR Virtual Conference: Thirteenth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2020 Oct 2-4. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(12 Suppl):Abstract nr PO-089.

Alteration of smoking habit at time of first diagnosis influences survival of patients with HNSCC.

The impact of smoking on survival in patients with squamous cell carcinoma of the head and neck is well established, despite some conflicting data in the literature. However, data on alterations of smoking habit following cancer diagnosis is sparse. In the present study, the effect of reduction of smoking compared with cessation on the course of disease was studied. Data from 643 patients with HNSCC from the tumor documentation registry of the Department of Otorhinolaryngology, Head and Neck Surgery of the Christian-Albrechts-University Kiel were collected and statistically analyzed, looking at pre- and post-treatment smoking habit and survival. Alteration of smoking at the first diagnosis of HNSCC led to a significantly beneficial effect on survival outcomes compared with continued smoking, without significant differences between reduction and cessation of smoking. Detailed analysis revealed that this effect was solely dependent on patients treated by surgery only. Lifelong non-smokers exhibited a significant survival advantage compared with active and former smokers, with no difference in survival between these last two groups. The positive influence of altered smoking habit following first time diagnosis on disease-specific survival paralleled the negative direct effect of active smoking on therapy, which is predominantly attributed to peritumoral tissue hypoxia leading to impaired efficacy of radiochemotherapy (RCT). In the present study cohort, the positive effect of smoking habit alterations were primarily observed in patients treated by surgery only instead of RCT, possibly due to fewer perioperative complications. These findings indicated that patients should be encouraged to at least minimize smoking following cancer diagnosis. Furthermore, for survival estimates and therapy planning, former smokers should be considered as active smokers.

LINC00958 and HOXC13-AS as key candidate biomarkers in head and neck squamous cell carcinoma by integrated bioinformatics analysis.

BackgroundHead and neck squamous cell carcinoma (HNSCC) is a malignant tumor with a strong tendency for metastasis and recurrence. Finding effective biomarkers for the early diagnosis of HNSCC is critical for the early treatment and prognosis of patients.MethodsRNA sequencing data including long non-coding RNAs (lncRNAs), messenger RNA (mRNAs) and microRNAs (miRNAs) of 141 HNSCC and 44 adjacent normal tissues were obtained from the TCGA. Differentially expressed genes were analyzed using the R package DESeq. GO terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted. A competing endogenous RNAs (ceRNA) network was constructed. The most differentially expressed genes in the main ceRNA network were chosen for nasopharyngeal carcinoma (NPC) cell lines and NPEC2 Bmi-1 cell line verification. A receiver operating characteristic (ROC) curve was constructed for 141 specimens of HNSCC tissues from 44 control samples.ResultsIn our study, 79 HNSCC-associated abnormally expressed lncRNAs , 86 abnormally expressed miRNAs and 324 abnormally expressed mRNAs were identified. The public microarray results showed that LINC00958 and HOXC13-AS expression levels were upregulated in HNSCC tissues compared with the adjacent normal tissues in this study (p < 0.0001). LINC00958 and HOXC13-AS expression levels in NPC cell lines were higher than those in the NPEC2 Bmi-1 cell line (p < 0.05). The results showed that the area under the ROC curve (AUC) of LINC00958 reached up to 0.906 at a cutoff value of 7.96, with a sensitivity and specificity of 80.85% and 90.91%, respectively. The AUC of HOXC13-AS reached up to 0.898 at a cutoff value of 0.695, with sensitivity and specificity values of 86.23% and 83.78%, respectively.ConclusionThe current study indicates that LINC00958 and HOXC13-AS are new candidate diagnostic biomarkers for HNSCC patients.

Logistic regression prediction model identify type 2 diabetes mellitus as a prognostic factor for human papillomavirus-16 associated head and neck squamous cell carcinoma.

BackgroundHPV-16–positive HNSCC and HPV-16–negative HNSCC have different clinical factors, representing distinct forms of cancers. The study aimed to identify patient-specific factors for HPV-16-positive HNSCC based on baseline clinical data.MethodFactors associated with HPV-16-positive HNSCC were identified using the data from 210 patients diagnosed with HNSCC at University College of London Hospital between January 1, 2003, and April 30, 2015, inclusive. A series of models were developed using logistic regression methods, and the overall model fit was compared using Akaike Information Criterion. Survival analysis was carried with Cox proportional hazards model for survival-time outcomes. The survival time for individual patients was defined as the time from diagnosis of HNSCC to the date of death from any cause. For patients who did not die, they were censored at the end of study on April 30, 2015.ResultsOf the 210 patients, 151 (72%) were found to have HPV-16-positive HNSCC. The logistic regression model showed that the prevalence of developing HPV-16-positive HNSCC was 3.79 times higher in patients with Type 2 Diabetes Mellitus (T2DM) (odd ratio [OR], 3.79; 95% CI, 1.70–8.44) than in those without T2DM, and 8.84 times higher in patients with history of primary HNSCC (OR, 8.84; 95% CI, 2.30–33.88) than in those without a history of primary HNSCC. HPV-16–positive HNSCC was also observed more in tonsils (OR, 4.02; 95% CL, 1.56–10.36) and less in non-alcohol drinker’s oral cavity (OR, 0.14; 95% CI, 0.03–0.56). Furthermore, individual patients were followed-up for 1 to 13 years (median of 1 year). Patients with HPV-positive HNSCC had a median survival of 5 years (95% CI, 2.6–7.3 years). Among HPV-16–positive HNSCC cohort, T2DM was a risk for poorer prognosis (hazard ratio, 2.57; 95% Cl, 1.09–6.07), and had lower median survival of 3 years (95% CI, 1.8–4.1 years), as compared to 6 years (95% CI, 2.8–9.1 years) in non-T2DM.ConclusionsPatient-specific factors for HPV-positive HNSCC are T2DM, history of primary HNSCC and tonsillar site. T2DM is associated with poorer prognosis. These findings suggest that it might be beneficial if routine HPV-16 screening is carried out in T2DM patients which can provide better therapeutic and management strategies.

Identification of SERPINE1, PLAU and ACTA1 as biomarkers of head and neck squamous cell carcinoma based on integrated bioinformatics analysis

BackgroundHead and neck squamous cell carcinoma (HNSCC) is the six leading cancer by incidence worldwide. The 5-year survival rate of HNSCC patients remains less than 65% due to lack of symptoms in the early stage. Hence, biomarkers which can improve detection of HNSCC should improve clinical outcome.MethodsGene expression profiles (GSE6631, GSE58911) and the Cancer Genome Atlas (TCGA) HNSCC data were used for integrated bioinformatics analysis; the differentially expressed genes (DEGs) were then subjected to functional and pathway enrichment analysis, protein–protein interaction (PPI) network construction. Subsequently, module analysis of the PPI network was performed and overall survival (OS) analysis of hub genes in subnetwork was studied. Finally, immunohistochemistry was used to verify the selected markers.ResultsA total of 52 up-regulated and 80 down-regulated DEGs were identified, which were mainly associated with ECM–receptor interaction and focal adhesion signaling pathways. Importantly, a set of prognostic signatures including SERPINE1, PLAU and ACTA1 were screened from DEGs, which could predict OS in HNSCC patients from TCGA cohort. Experiment of clinical samples further successfully validated that these three signature genes were aberrantly expressed in the oral epithelial dysplasia and HNSCC, and correlated with aggressiveness of HNSCC patients.ConclusionsSERPINE1, PLAU and ACTA1 played important roles in regulating the initiation and progression of HNSCC, and could be identified as key biomarkers for precise diagnosis and prognosis of HNSCC, which will provide potential targets for clinical therapies.

PO-080 Prevalence of human papilloma virus and its phylogenetic analysis in patients with head and neck squamous cell carcinoma in pakistan

IntroductionHNSCCs are characterised by distinct phenotypic, aetiological, biological and clinical heterogeneity. It is a heterogeneous disease that can be roughly divided into human papillomavirus (HPV) positive and negative. HPV is a recently identified causative agent for a subset of head and neck cancers, primarily in the oropharynx but very little is known about the prevalence of specific HPV types in Pakistan.Material and methodsThis cross sectional study was carried out in the Department of Immunology, University of Health Sciences, Lahore, Pakistan after approval from the Ethical Review Committee and Advanced Studies and Research Board, UHS Lahore. After an informed consent, tissue sections from a total of 72 patients with HNSCC were collected. The diagnosis of HNSCC was confirmed again on H and E staining and DNA was extracted from formalin fixed paraffin embedded tissue sections using DNA extraction kit. For the detection of HPV DNA, two universal primer sets GP5/GP6 and MY9/MY11 were used in the polymerase chain reaction. The samples, found to be positive for HPV on PCR, were sequenced for the phylogenetic analysis of HPV.Results and discussionsAmong 72 cases of HNSCC, only 5 (14.4%) cases were found to be positive for HPV by using universal primers sets. The prevalence of HPV in HNSCC in many international studies from Europe and USA have documented a prevalence of 5%–50% in HNSCC cases. After confirming the presence of HPV in HNSCC cases, positive DNA samples have been transported to a well-known international laboratory for gene sequencing and molecular diagnosis. As soon as possible, we will receive the sequencing data, we will sketch a phylogenetic tree and further subtyping of HPV and the further results will be presented.ConclusionThe findings of current study will be helpful in launching public health awareness and future vaccination programs against specific HPV types in Pakistan to lessen the burden of HPV related HNSCC.

Brain metastasis from squamous cell carcinoma of the head and neck: a review of the literature in the genomic era.

Squamous cell carcinoma of the head and neck (HNSCC) affects nearly 500,000 individuals globally each year. With the rise of human papillomavirus (HPV) in the general population, clinicians are seeing a concomitant rise in HPV-related HNSCC. Notably, a hallmark of HPV-related HNSCC is a predilection for unique biological and clinical features, which portend a tendency for hematogenous metastasis to distant locations, such as the brain. Despite the classic belief that HNSCC is restricted to local spread via passive lymphatic drainage, brain metastases (BMs) are a rare complication that occurs in less than 1% of all HNSCC cases. Time between initial diagnosis of HNSCC and BM development can vary considerably. Some patients experience more than a decade of disease-free survival, whereas others present with definitive neurological symptoms that precede primary tumor detection. The authors systematically review the current literature on HNSCC BMs and discuss the current understanding of the effect of HPV status on the risk of developing BMs in the modern genomic era.

Ectonucleotidase CD39 expression in regional metastases in head and neck cancer

Introduction: CD39 is the rate-limiting enzyme in the generation of immunosuppressive adenosine and its expression and activity are significant in tumor progression. Squamous cell carcinoma of the head and neck (HNSCC) shows an overall poor prognosis due to high local recurrence rates and early metastatic spread.Material and methods: Primary tumor specimens and lymph node specimens harvested during neck dissection of 65 patients with a diagnosis of HNSCC were subjected to immunohistochemical and H-score analysis of CD39 expression. Demographics, histopathology and subsequent outcome were analyzed.Results: The primary cancer was squamous cell carcinoma in all patients (male/female 55:10). H-score for CD39 expression in the primary lesion and metastatic lymph nodes was significantly higher in advanced compared to early stages with no significant differences among different tumor locations. High intratumoral and intrametastatic CD39 expression was associated with an inferior patients’ overall survival at a mean follow-up of 83.4 months (6–204 months).Conclusion: CD39 expression in HNSCC correlated positively with tumor stage and appears to predict poor prognosis. Therefore, CD39 expression in primary lesions and metastatic lymph nodes seems to identify patients at high risk in HNSCC of all tumor sites. Immunotherapeutic approaches targeting CD39 might be promising for this patient population.

Recent advances in early diagnosis of head and neck cancer in precision medicine era

Head and neck squamous cell carcinoma, HNSCC, has high morbidity and mortality. Even in America, more than 1/2 to 2/3 patients have been diagnosed at advanced stage. So, it's urgent to find ways to diagnose HNSCC earlier and foresee the curative effect. With the achievement of Next-Generation Sequencing, researchers are trying to develop early diagnostic technology from a new perspective, such as personal genomics, proteomics, metabolomics and other related personal information. Here we reviewed the recent research in early diagnosis of HNSCC.