Development Of HNSCC Research Articles

Validation of biomarkers and immunotherapy in head and neck squamous cell carcinoma using bioinformatics and Mendelian randomization

Background To promote carcinogenesis through diverse molecular pathways involving dysregulation of gene expression and abnormalities. Methods We employed Mendelian randomization (MR) to uncover causal relationships between genetic factors and HNSCC. We used the Inverse Variance Weighted (IVW) method as the primary MR analysis, and validated the results through complementary approaches like MR-Egger regression, weighted median, and mode analyses. Results Our analysis identified 2210 genes that are differentially expressed in head and neck cancer (HNSCC) compared to normal tissues. Within the protein interaction network, the genes IL1B, CXCL8, CXCL1, and CCL2 stood out as central hubs. Further investigation revealed that these key genes are involved in important biological processes like skin development, wound healing, and fat metabolism. Notably, our Mendelian randomization analysis provided evidence for a causal relationship between the expression of the IL1B gene and the development of HNSCC. Conclusions Our analysis identified 5 key genes - IL1B, CXCL8, CXCL1, CCL2, and IL1B - that show significant changes in expression in head and neck cancer. These genes could serve as important new biomarkers to help diagnose this disease and track how it progresses over time. Importantly, these genes are involved in regulating the immune system, suggesting that the body’s immune response plays a critical role in head and neck cancer. This provides new avenues for future research to better understand the complex gene expression patterns underlying this type of cancer. Further investigation of these key genes and their regulatory networks could lead to important insights and potential new treatment approaches.

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.

An Investigation on the Genetic Alterations of the ALG3 Gene and Their Possible Correlation with the Development of HNSCC and LUSC

An Investigation on the Genetic Alterations of the ALG3 Gene and Their Possible Correlation with the Development of HNSCC and LUSC

Prognostic and clinicopathological significance of MRC2 expression in head and neck squamous cell carcinoma

BackgroundHead and neck squamous cell carcinoma (HNSCC) is one of the most aggressive types of cancers worldwide, with metastasis being the major cause of death. Recent research suggests that changes in the expression of MRC2 (mannose receptor, C-type 2) may play a role in the development and progression of various cancers; however, its expression pattern in HNSCC/ OSCC is unknown. This study aimed to elucidate the clinicopathological significance and prognostic role of MRC2 expression in HNSCC, including OSCC. Materials and methodsIn the present study, we assessed the potential roles of MRC2 in expression, prognostic value, immune infiltration and functional enrichment analysis in HNSCC patients by using different bioinformatics databases. We then validated MRC2 gene expression in 30 OSCC and adjacent normal tissue samples using quantitative reverse transcription PCR (RT-qPCR). ResultsMRC2 mRNA and protein expression were significantly upregulated in OSCC and HNSCC patients compared to that in adjacent normal tissues. Upregulated MRC2 expression was associated with poor overall survival. Increased MRC2 expression has also been linked to an aggressive clinicopathological features including advanced stages, grade, metastasis and HPV status. Interestingly, our in silico results strongly suggest that the MRC2 gene and protein interaction networks are associated with HNSCC development. Moreover, the tumor infiltration level was significantly correlated with HPV-negative HNSCC patients. ConclusionOur results suggest that MRC2 could be used as a novel prognostic marker and therapeutic target for HNSCC.

Influence of pathogens on host genome and epigenome in development of head and neck cancer.

Head and neck cancer (HNSCC) is a heterogeneous group of cancers, affecting multiple regions such as oral cavity, pharynx, larynx, and nasal region, each showing a distinct molecular profile. HNSCC accounts for more than 6 million cases worldwide, soaring mainly in the developing countries. The aetiology of HNSCC is complex and multifactorial, involving both genetic and environmental factors. The critical role of microbiome, which includes bacteria, viruses, and fungi, is under spotlight due to the recent reports on their contribution in the development and progression of HNSCC. This review focuses on the effect of opportunistic pathogens on the host genome and epigenome, which contributes to the disease progression. Drawing parallels from the host-pathogen interactions observed in other tumour types arising from the epithelial tissue such as colorectal cancer, the review also calls attention to the potential explorations of the role of pathogens in HNSCC biology and discusses the clinical implications of microbiome research in detection and treatment of HNSCC. Our understanding of the genomic effects of the microbes on the disease progression and the mechanistic insights of the host-pathogen interaction will pave way to novel treatment and preventive approaches in HNSCC.

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.

Roles of E2F family members in the diagnosis and prognosis of head and neck squamous cell carcinoma

BackgroundHead and neck squamous cell carcinoma (HNSCC) is the sixth most prevalent cancer worldwide. E2Fs are a group of transcription factors involved in the carcinogenesis and progression of various cancers. However, the exact roles of each member of E2F family in the development and progression of HNSCC are still unknown.MethodsRNASeq and clinical follow-up information were extracted from The Cancer Genome Atlas (TCGA). The expressions of E2Fs and their roles in HNSCC progression were explored using the R software and the cBioPortal database.ResultsOur results showed that the mRNA levels of E2Fs were significantly higher in HNSCC tumors than in normal tissues. E2F1, E2F3, E2F4, E2F6, and E2F7 were identified as reliable diagnostic markers. E2Fs (except for E2F3) expressions were closely related to the clinical features (excluding metastasis) of HNSCC. High E2F6 mRNA expression was an independent risk factor for the OS of female HNSCC patients. In addition, high E2F4 expression could lead to poor prognosis in HNSCC in both males and females, high expressions of E2F5, E2F6, and E2F7 were associated with poor OS of female HNSCC patients, while high E2F2 and E2F8 expressions were positively correlated with the OS of male HNSCC patients. Interestingly, E2Fs expressions had stronger associations with immune cell infiltrations in male HNSCC patients than in female HNSCC patients.ConclusionThe expressions of E2Fs were found to be correlated with the progression of HNSCC. E2F1, E2F3, E2F4, E2F6, and E2F7 could be good diagnostic molecules for HNSCC. In addition, E2F6 was an independent risk factor for the prognosis of female HNSCC patients.

STAT1 is regulated by TRIM24 and promotes immunosuppression in head and neck squamous carcinoma cells, but enhances T cell antitumour immunity in the tumour microenvironment

BackgroundHead and neck squamous cell carcinoma (HNSCC) is a significant problem and is frequently resistant to current treatments. STAT1 is important in anti-tumour immune responses against HNSCC. However, the role of STAT1 expression by tumour cells and its regulation during HNSCC is unclear.MethodsWe determined the effects of STAT1 inhibition on tumour development and immunity in CAL27 and UMSCC22A HNSCC cell lines in vitro and in a HNSCC carcinogen-induced model in vivo.ResultsSTAT1 siRNA knockdown in human HNSCC cells impaired their proliferation and expression of the immunosuppressive marker PD-L1. Stat1-deficient mice displayed increased oral lesion incidence and multiplicity during tumour carcinogenesis in vivo. Immunosuppressive markers PD-1 in CD8+ T cells and PD-L1 in monocytic MDSCs and macrophages were reduced in oral tumours and draining lymph nodes of tumour-bearing Stat1-deficient mice. However, STAT1 was required for anti-tumour functions of T cells during HNSCC in vivo. Finally, we identified TRIM24 to be a negative regulator of STAT1 that plays a similar tumorigenic function to STAT1 in vitro and thus may be a potential target when treating HNSCC.ConclusionOur findings indicate that STAT1 activity plays an important role in tumorigenicity and immunosuppression during HNSCC development.

Head and Neck Cancer Susceptibility and Metabolism in Fanconi Anemia.

Simple SummaryGermline loss-of-function mutations in any of over 20 known Fanconi anemia (FA) genes cause the disorder FA. The FA pathway plays a key role in the repair of DNA interstrand crosslinks (ICLs) and stabilization of stalled replication fork. Epidemiological studies have shown that persons with FA have a uniquely increased susceptibility to squamous cell carcinomas (SCCs) and increased human papilloma virus (HPV) prevalence. Genomic and transcriptomic studies are informative, but there is a paucity of data regarding metabolic deregulation in FA deficient systems. Metabolism is an essential component of cancer development and has significant impact on therapeutic outcomes. Therefore, a deeper understanding of the metabolic architecture of FA may aid in our understanding of SCC development, and allow for the discovery of metabolism-based diagnostic biomarkers and new therapeutic targets. In this review, we discuss known metabolic consequences of FA pathway loss on cancer pathogenesis, and in particular in normal and transformed keratinocytes—the cells of origin for SCC.Fanconi anemia (FA) is a rare inherited, generally autosomal recessive syndrome, but it displays X-linked or dominant negative inheritance for certain genes. FA is characterized by a deficiency in DNA damage repair that results in bone marrow failure, and in an increased risk for various epithelial tumors, most commonly squamous cell carcinomas of the head and neck (HNSCC) and of the esophagus, anogenital tract and skin. Individuals with FA exhibit increased human papilloma virus (HPV) prevalence. Furthermore, a subset of anogenital squamous cell carcinomas (SCCs) in FA harbor HPV sequences and FA-deficient laboratory models reveal molecular crosstalk between HPV and FA proteins. However, a definitive role for HPV in HNSCC development in the FA patient population is unproven. Cellular metabolism plays an integral role in tissue homeostasis, and metabolic deregulation is a known hallmark of cancer progression that supports uncontrolled proliferation, tumor development and metastatic dissemination. The metabolic consequences of FA deficiency in keratinocytes and associated impact on the development of SCC in the FA population is poorly understood. Herein, we review the current literature on the metabolic consequences of FA deficiency and potential effects of resulting metabolic reprogramming on FA cancer phenotypes.

First-degree family history of cancer can be a potential risk factor among head and neck cancer patients in an isolated Mizo tribal population, northeast India

BackgroundHead and Neck Squamous Cell Carcinoma (HNSCC) is commonly associated with tobacco and alcohol consumption. The aim of this study is to find the association between the risk factors with HNSCC in a small tribal population of Mizoram, North-East India. MethodsData were obtained through consented questionnaires and Logistic Regression was used to calculate the Odds Ratio (OR) between the parameters and HNSCC. ResultSignificant association was observed for smoking and alcohol with an OR of 6.703 and 4.527, respectively. The OR was found to increase with increase in smoking and alcohol consumption. Majority of the patients consumed local made alcohol and smoked the local made cigarettes known as Zozial. Moreover, the First-Degree Family History of Cancer showed a significant OR of 1.921 (95% CI: 1.040–3.547). ConclusionRegardless of the duration of smoking or alcohol consumption, Family History of Cancer might influence the risk of HNSCC. Further screening is essential to evaluate the potential role of germline mutational effect on development of HNSCC in the population.

MicroRNAs: Their Role in Metabolism, Tumor Microenvironment, and Therapeutic Implications in Head and Neck Squamous Cell Carcinoma.

Simple SummaryHead and neck squamous cell carcinoma (HNSCC), which arises from the oral epithelium, is one of the most common cancers worldwide. Despite excellent diagnosis and treatment improvements, the mortality rate associated with HNSCC is still extremely high. Current data suggest that dysregulation of exosomes and metabolic abnormalities are involved in the initiation and progression of HNSCC. Thus, approaches for targeting exosomes in the tumor microenvironment and metabolic reprogramming pathways represent potential therapeutic strategies. Moreover, some miRNAs are thought to have significant functions in regulating the progression of HNSCC. The present article aims to summarize the current knowledge concerning the important miRNAs in both exosomes and cancer metabolism, as well as discuss future perspectives regarding their future diagnostic potential and treatment recommendations.MicroRNAs (miRNAs) are endogenous small non-coding RNA molecules that negatively regulate gene expression by binding to target mRNAs. Deregulated miRNAs can act as either oncogenic miRNAs or tumor suppressor miRNAs in controlling proliferation, differentiation, apoptosis, metastasis, epithelial–mesenchymal transition, and immune responses, which are all involved in the carcinogenesis process of HNSCC. Recent findings have shown that metabolic reprogramming is an important hallmark of cancer, which is necessary for malignant transformation and tumor development. Some reprogrammed metabolisms are believed to be required for HNSCC against an unfavorable tumor microenvironment (TME). The TME is composed of various cell types embedded in the altered extracellular matrix, among which exosomes, secreted by cancer cells, are one of the most important factors. Tumor-derived exosomes reshape the tumor microenvironment and play a crucial role in cell-to-cell communication during HNSCC development. Exosomes encapsulate many biomolecules, including miRNAs, circulate in body fluids, and can transmit intercellular regulatory messages to nearby and distant sites, which indicates that exosomal miRNAs have the potential to become non-invasive biomarkers. This review aims to clarify the functions of diverse miRNAs in HNSCC metabolic reprogramming and tumor-derived exosomes. In addition, it also emphasizes the potential role of miRNA as a biomarker in the diagnosis, prognosis, and treatment of HNSCC cancer.

Prognostic potential of Akt, pAkt(Ser473) and pAkt(Thr308) immunoreactivity in relation to HPV prevalence in head and neck squamous cell carcinoma patients

BackgroundThe prognosis of squamous cell carcinoma of head and neck (HNSCC) patients remains relatively poor over the last years. Tobacco, alcohol and active human papillomavirus (HPV) infection are involved in HNSCC development. Akt is a serine-threonine protein kinase with main phosphorylation sites at Thr308 and Ser473, which are critical to generate a high level of Akt activity. Materials and methodsThe aim of the study was to compare the expression and prognostic potential of total Akt and its 2 phosphorylated forms – pAkt(Ser473) and pAkt(Thr308) in relation to HPV status in HNSCC patients. The expression levels of proteins were assessed immunohistochemically. To select independent prognostic factors univariate and multivariate analyses with Cox proportional regression model were performed. ResultsAmong HNSCC with active HPV16 infection significantly more tumors with high Akt (67.86%, p = 0.026) and low pAkt(Ser473) (64.29%, p = 0.000) expressions were found as compared to those with HPV negativity, while there was no significant difference in the pAkt(Thr308) expression level between HPV positive and negative tumors (p = 0.359). In the whole group of HNSCC patients independent favorable prognostic factors were low T stage, low pAkt(Thr308) expression, HPV16 active infection presence (for OS and DFS) and female gender (for OS only). ConclusionsOur results indicate an important role of pAkt(Thr308) as prognostic biomarker for HNSCC patients. There is a high probability that using Akt inhibitors would improve therapeutical benefits and treatment effectiveness, especially in HNSCC patients with high expression of pAkt.

Abstract LB194: Analysis of Hippo/YAP abnormalities in head and neck cancer

Abstract Introduction: Squamous cell carcinomas (HNSCC) comprise more than 95% of all head and neck cancers. Over the past decades the incidence and mortality rates of HNSCC have been steadily increasing worldwide. HNSCCs account for 6% of all human cancers and for 3% of all cancers in the United States. Leukoplakia, also known as smoker's keratosis are described as non-cancerous, pre-malignant inflammatory patches is a major risk factor in the development of HNSCC. In fact, up to 31% of all leukoplakias are believed to transform into HNSCC. Our interest into the molecular contributions to head and neck carcinogenesis has led us to follow up intriguing findings on the contributions of Yes Associated Protein (YAP) pathways that were found to upregulate proliferation, invasion/migration, and survival in head and neck cancer. More recent studies have found that Hippo/Yap activation occurs in many human cancers as well. Materials and Methods: We performed several in silico analyses of available human cancer genetic data. We interrogated data from various Head and Neck Cancer studies. Specifically, we included data from the Recurrent and Metastatic Head & Neck Cancer (MSKCC, JAMA Oncol 2016), Head and Neck Squamous Cell Carcinoma (Broad, Science 2011), Head and Neck Squamous Cell Carcinoma (Johns Hopkins, Science 2011), Head and Neck Squamous Cell Carcinoma (TCGA, PanCancer Atlas), and the Oral Squamous Cell Carcinoma (MD Anderson, Cancer Discov 2013) using CBioPortal web tool. We explored factors responsible for Hippo pathway activation via loss of negative regulators or amplification of Hippo drivers. Results: We found many negative regulators, such as TAOK1, LATS1/2, among others to be mutated, exclusive of FAT1 loss, a known event in HNSCC. We also found amplification of YAP1, a driver of the Hippo pathway. We saw a loss of FAT1 in 22% of tumors, mutations in TAOK1 and LATS1/2 were seen in over 5% of tumors, exclusive of other mutations or gene expression derangements. YAP1 amplification was observed in up to 5% of tumors. Conclusions: These findings substantiate earlier findings where the YAP pathway was found to be upregulated in head and neck cancer subsets. Intriguingly, the loss of negative regulators or amplification of oncogenes, such as YAP1, likely contribute to Hippo/YAP upregulation. Since they are important pathways that may be activated at the crux of oral carcinoma in situ/invasion, findings from our in silico interrogation shed light on genetic abnormalities that probably occur earlier in the carcinogenesis spectrum. Further interrogation of leukoplakia preclinical models or data sets could shed additional light on contributions of Hippo/YAP to oral carcinogenesis. Citation Format: German Velez Reyes, Beverly Wuertz, Frank G. Ondrey. Analysis of Hippo/YAP abnormalities in head and neck cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB194.

The m6A RNA Modification Quantity and mRNA Expression Level of RNA Methylation-Related Genes in Head and Neck Squamous Cell Carcinoma Cell Lines and Patients.

RNA methylation at the nitrogen sixth of adenosine (m6A, N6-methyladenosine) is the most abundant RNA modification which plays a crucial role in all RNA metabolic aspects. Recently, m6A modification has been assigned to mediate the biological processes of cancer cells, but their significance in HNSCC development is still poorly described. Thus, the main aim of this study was to globally quantify m6A modification by the mass spectrometry approach and determine the mRNA expression level of selected m6A RNA methyltransferase (METTL3), demethylase (FTO), and m6A readers (YTHDF2, YTHDC2) in 45 HNSCC patients and 4 cell lines (FaDu, Detroit 562, A-253 and SCC-15) using qPCR. In the results, we have not observed differences in the global amount of m6A modification and the mRNA level of the selected genes between the cancerous and paired-matched histopathologically unchanged tissues from 45 HNSCC patients. However, we have found a positive correlation between selected RNA methylation machinery genes expression and m6A abundance on total RNA and characterized the transcript level of those genes in the HNSCC cell lines. Moreover, the lack of global m6A differences between cancerous and histopathologically unchanged tissues suggests that m6A alterations in specific RNA sites may specifically influence HNSCC tumorigenesis.

Bioinformatics analysis of the expression and role of microRNA-221-3p in head and neck squamous cell carcinoma

BackgroundHead and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide, associated with a high rate of morbidity and mortality. However, the target genes of miR-221-3p and the underlying mechanism involved in HNSCC are still not clear. Therefore, in the current study, we studied the role of miR-221-3p in the HNSCC.MethodsTissues collected from 48 control and 21 HNSCC patients were processed to check the differential expression of miR-221-3p by RT-qPCR. Overexpression of microRNA-221-3p (miR-221-3p) is significantly correlated to the onset and progression of HNSCC. We also conducted the meta-analysis of the cancer literature from the cancer genome atlas (TCGA) and the Gene Expression Omnibus (GEO) database to estimate the expression of miR-221-3p in HNSCC. The miR-221-3p target genes in the HNSCC were predicted with the miRWalk and TCGA databases, and functionally annotated via the Gene Ontology. Finally, Spearman’s analysis was used to determine the role of the related target genes in important pathways involved in the development of HNSCC.ResultsWe observed a significantly higher expression of miR-221-3p in HNSCC compared to the normal with a summary receiver operating characteristic (sROC) of 0.86(95% Cl: 0.83,0.89). The KEGG and GO comprehensive analysis predicted that miR-221-3p might be involved in the development of HNSCC through the following metabolic pathways, viz. Drug metabolism - cytochrome P450 UGT1A7 and MAOB may be important genes for the role of miR-221-3p.ConclusionBased on bioinformatics analysis, our results indicate that miR-221-3p may be used as a non-invasive and hypersensitive biomarker in the diagnosis. Thus, it can be concluded that miR-221-3p may be an extremely important gene locus involved in the process of the deterioration and eventual tumorigenesis of HNSCC. Hopefully, additional work will validate its usefulness as a target for future clinical research.

Development and validation of a RNA binding protein-associated prognostic model for head and neck squamous cell carcinoma.

Evidence shows that defects in RNA-binding proteins (RBPs) are closely related to the occurrence and development of HNSCC. We obtained 502 tumors and 44 normal samples from the TCGA database, among which 190 differentially expressed RBPs were screened. Finally, a prognostic model containing nine RBPs (CELF2, CPEB1, DDX39B, EIF3L, EZH2, KHDRBS3, RNASE10, RNASE3 and SIDT1) was produced. Further analysis showed that the overall survival rate in the high-risk group was lower than that in the low-risk group. The area under the ROC curve (AUC) in the training and testing groups was significant (3-year AUC, 0.735 vs 0.796; 5-year AUC, 0.821 vs 0.804). In addition, a comprehensive analysis of nine identified RBPs showed that most of them were related to the OS of HNSCC patients, and three of them (CELF2, EZH2, and SIDT1) were differentially expressed in HNSCC and control tissues at the protein level. In addition, our data revealed that the identified RBPs are highly interconnected, with high frequency copy number changes in HNSCC samples. GSEA indicated that the abnormal biological processes related to RNA and the activation of some classical tumor signaling pathways were important driving forces for the development of HNSCC. Our results provide novel insights into the pathogenesis of HNSCC, among which nine RBP markers have potential application value in clinical decision-making and individualized treatment of HNSCC.

Evaluation of Pharyngeal Background Mucosa in Patients with Superficial Hypopharyngeal Carcinoma

Transoral surgery (TOS) has become increasingly popular for patients with superficial hypopharyngeal squamous cell carcinoma (SCC). However, the number of patients in whom metachronous multiple SCC of the head and neck (HNSCC) occurs has also increased. In this study, we investigated whether multiple lugol-voiding lesions (LVLs) in the pharyngeal background mucosa observed during TOS would be a biomarker of metachronous HNSCC. Retrospective study. We examined 362 patients who underwent TOS for superficial hypopharyngeal carcinoma. Endoscopic images were reviewed in a blinded fashion by two endoscopists. LVLs in the pharyngeal mucosa were graded as follows: A, no lesions; B, 1 to 4 lesions; and C, ≥5 lesions per endoscopic view. Cumulative incidence curves of secondary HNSCC in the groups of grades A, B, and C revealed 3-year incidence rates of 14.4%, 18.8%, and 29.3%, respectively (P=.001 for A vs. C and P=.002 for B vs. C). Cumulative incidence curves of third HNSCC in the groups of grades A, B. and C revealed 5-year incidence rates of 3.9%, 9.8%, and 19.6%, respectively (P=.001 for A vs. C and P=.006 for B vs. C). Cumulative incidence curves of fourth HNSCC in the groups of grades A, B, and C revealed 7-year incidence rates of 0%, 2.3%, and 13.2%, respectively (P=.025 for A vs. C and P=.009 for B vs. C). Multiple LVLs in the pharyngeal mucosa increase the risk of development of metachronous multiple HNSCC. 3 (nonrandomized, controlled cohort/follow-up study) Laryngoscope, 131:2036-2040, 2021.

Detection of Field Cancerization in The Clinically Normal Oral Mucosa of Cannabis and Cigarette Smokers

Background: Head and neck cancer represents one of the most common cancer types. Numerous riskfactors are implicated in the development of HNSCC. In our study we aimed to investigate the changes thatoccur in the oral mucosa of cannabis smokers and compare them to cigarette smokers and non-smokers.Methods: Three groups of subjects were included in our study (33 in each group); cannabis and cigarettesmokers, cigarette smokers and non-smokers. The biopsies were examined by routine H&E techniques andimmunohistochemical expression of p53.Conclusion: The present work demonstrated evident dysplastic and pre-dysplastic changes in H&E stainedsections. The changes were detected histologically in both smoker groups. These changes were morepronounced in cannabis smokers’ group than cigarette smokers’ group. Moreover, p53 immunostaining washigher for cigarette smokers’ group than cannabis smokers’ group. It is evident that cigarette smoking andcannabis smoking results in field changes in the oral mucosa which are detectable in tissue sections. Thesechanges, such as hyperplasia and dysplasia are more detectable in H&E sections of cannabis smokers thancigarette smokers. However, positive p53 immunoexpression was lower in cannabis smokers than cigarettesmokers.

Differentiation and TNM staging of Oral Squamous Cell Carcinoma

Head and neck carcinoma has great regional distribution in sub sites with distinct anatomical and physiological variation. Alcohol consumption, excessive smoking and hematopoetic stem cell transplantation are common risk factors for head and neck carcinoma. Alcohol consumption & tobacco chewing has a strong relationship with the development of oral, larynx, pharynx cancers. A typical HNSCC has a superficial changes such as scaling, crusting, presence of cutaneous horn and ulceration. The stage of disease is apparent in the form of symptoms of head and neck carcinoma. Genetic alterations accumulation is source of initiation and development of carcinogenic cells. The development of HNSCC is based on many factors. Its underlying mechanism is based on the genetic and molecular changes along with repetitive injury to the epithelial layer by external stimuli. Different types of smoking such as Cigar, Pipe, Huqqa etc exist in sub-continent and smoke from these sources contain toxic agents like tar and carbon monoxide. Mouth cavity and lungs are the places where tar being sticky substance deposits and cause many respiratory and oral diseases.

Overexpression of wild type or a Q311E mutant MB21D2 promotes a pro-oncogenic phenotype in HNSCC.

Cadherin-mediated cell-cell contacts regulated by intracellular binders play critical roles in tissue homeostasis and tumorigenesis. Here, we screened mutational profiles of 312 annotated genes involved in cadherin binding in human squamous cell carcinomas and found MB21D2 to carry a unique recurrent Q311E mutation. MB21D2 overexpression was also frequently found in head and neck cancer (HNSCC) and was associated with poor clinical outcomes. Cell-based characterizations revealed pro-oncogenic roles for MB21D2 wild-type (WT) and its Q311E mutant (Q311E) in cell proliferation, colony formation, sphere growth, and migration/invasion by promoting epithelial-mesenchymal transition. Conversely, MB21D2 knockdown in MB21D2-overexpressing cells resulted in cell growth arrest and apoptosis. Xenograft tumor models with Q311E-expressing cells formed larger and more aggressive lesions, compared to models with WT-MB21D2-expressing cells or an empty vector. Transcriptome and protein interactome analyses revealed enrichment of KRAS signaling by MB21D2 expression. Immunoblotting confirmed RAS elevation, along with upregulation/phosphorylation of PI3K, AKT, and CREB. Blocking RAS signaling in MB21D2-expressing cells by manumycin significantly reduced cell growth and survival. Our study thus defined RAS signaling-dependent pro-oncogenic roles for MB21D2 overexpression and Q311E MB21D2 expression in HNSCC development.