Pathogenesis Of Head And Neck Squamous Cell Carcinoma Research Articles

P.200 Immunological substrates of depressive symptoms in patients with severe obesity

While tobacco smoking is a well-known risk factor for head and neck squamous cell carcinoma (HNSCC), the molecular mechanisms underlying tobacco-induced HNSCC remain unclear. This study sought to comprehensively identify microRNA (miRNA) alterations and evaluate their clinical relevance in smoking-induced HNSCC pathogenesis and progression.Using small RNA-sequencing data and clinical data from 145 HNSCC patients, we performed a series of differential expression and correlation analyses to identify a panel of tobacco-dysregulated miRNAs associated with key clinical characteristics in HNSCC. We then examined the expression patterns of these miRNAs in normal epithelial cell lines following exposure to cigarette smoke extract.Our analyses revealed distinct panels of miRNAs to be dysregulated with smoking status and associated with additional clinical features, including tumor stage, metastasis, anatomic site, and patient survival. The differential expression of key miRNAs, including miR-101, miR-181b, miR-486, and miR-1301, was verified in cigarette-treated epithelial cell lines, suggesting their potential roles in the early development of smoking-related HNSCCs.Specific alterations in miRNA expression may be traced to tobacco use and are associated with important HNSCC clinical characteristics. Future studies of these miRNAs may be valuable for furthering the understanding and targeted treatment of smoking-associated HNSCC.

Mutation signatures in head and neck squamous cell carcinoma : Pathogenesis and therapeutic potential

The pathogenesis of head and neck squamous cell carcinoma (HNSCC) is acomplex and multistage process which results from the interaction of exogenous and endogenous cellular processes. Each of these processes leaves acharacteristic pattern of mutations on the tumor genome, aso-called mutational signature. The subject of current studies is to decipher specific signatures of mutational processes operating during HNSCC pathogenesis and to address their prognostic value. Computational analysis of genomic sequencing data by The Cancer Genome Atlas (TCGA) revealed mutational signatures1, 2, 4, 5,7, and 13 as the main players in HNSCC pathogenesis. Signature16 was first discovered in human papillomavirus (HPV)-negative oral and oropharyngeal tumors. In many studies, an association of signature16 with alcohol and tobacco consumption as well as with an unfavorable prognosis was described.

Regulation of NFκB Signalling by Ubiquitination: A Potential Therapeutic Target in Head and Neck Squamous Cell Carcinoma?

Simple SummaryHead and neck cancer is the sixth most common cancer worldwide and typically caused by smoking and alcohol consumption, or infection with Human Papillomavirus (HPV). Currently, treatment options include surgery, radiotherapy, and/or chemotherapy. However, whilst the survival rate in HPV+ cancer patients is better than those without HPV, survival rates have not improved greatly in recent years, and recurrence rates are high. Ubiquitination is a critical regulatory mechanism that can promote the activation or termination of signal cascades, such as the NFκB pathway, which plays an important role in the pathogenesis and therapeutic resistance of head and neck cancer. In this review, we discuss how NFκB signalling is regulated by ubiquitination and how the ubiquitin pathway is deregulated in head and neck cancer, highlighting how the this pathway may be targeted to inhibit NFκB signalling.Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide, with over 600,000 cases per year. The primary causes for HNSCC include smoking and alcohol consumption, with an increasing number of cases attributed to infection with Human Papillomavirus (HPV). The treatment options for HNSCC currently include surgery, radiotherapy, and/or platinum-based chemotherapeutics. Cetuximab (targeting EGFR) and Pembrolizumab (targeting PD-1) have been approved for advanced stage, recurrent, and/or metastatic HNSCC. Despite these advances, whilst HPV+ HNSCC has a 3-year overall survival (OS) rate of around 80%, the 3-year OS for HPV− HNSCC is still around 55%. Aberrant signal activation of transcription factor NFκB plays an important role in the pathogenesis and therapeutic resistance of HNSCC. As an important mediator of inflammatory signalling and the immune response to pathogens, the NFκB pathway is tightly regulated to prevent chronic inflammation, a key driver of tumorigenesis. Here, we discuss how NFκB signalling is regulated by the ubiquitin pathway and how this pathway is deregulated in HNSCC. Finally, we discuss the current strategies available to target the ubiquitin pathway and how this may offer a potential therapeutic benefit in HNSCC.

Traditional Herbal Medicine Mediated Regulations during Head and Neck Carcinogenesis.

Head and neck squamous cell carcinoma (HNSCC) is one of the most prevalent neoplasms worldwide. It is well recognized that environmental challenges such as smoking, viral infection and alcohol consumption are key factors underlying HNSCC pathogenesis. Other than major clinical interventions (e.g., surgical resection, chemical and radiotherapy) that have been routinely practiced over years, adjuvant anticancer agents from Traditional Herbal Medicine (THM) are proposed, either alone or together with conventional therapies, to be experimentally effective for improving treatment efficacy in different cancers including HNSCCs. At a cellular and molecular basis, THM extracts could modulate different malignant indices via distinct signaling pathways and provide better control in HNSCC malignancy and its clinical complications such as radiotherapy-induced xerostomia/oral mucositis. In this article, we aim to systemically review the impacts of THM in regulating HNSCC tumorous identities and its potential perspective for clinical use.

Determining the molecular landscape and impact on prognosis in HPV-associated head and neck cancer

BackgroundHuman papillomavirus (HPV) associated head and neck squamous cell carcinoma (HNSCC) has a better prognosis than HNSCC due to other risk factors. However, there is significant heterogeneity within HPV-associated HNSCC and 25% of these patients still do poorly despite receiving aggressive therapy. We currently have no good molecular tools to differentiate and exclude this “high-risk” sub-population and focus on “low-risk” patients for clinical trials. This has been a potential barrier to identifying successful de-escalation treatment strategies in HPV-associated HNSCC. We conducted an analysis of molecular markers with a well-known role in the pathogenesis of HPV-associated HNSCC and hypothesized that these markers could help independently predict recurrence and prognosis in these patients and therefore help identify at the molecular level “low-risk” patients suitable for de-escalation trials.MethodsWe analyzed 24 tumor specimens of patients with p16+ HNSCC who underwent definitive resection as primary treatment. Tissue microarray (TMA) was generated from the 24 pathology blocks and immunohistochemistry (IHC) was performed using highly specific antibodies for our chosen biomarkers (PI3K-PTEN, AKT pathway, mTOR, 4EBP1, S6, and pAMPK, ERCC-1). Transcriptome data was also obtained for 7 p16+ HNSCC patients from The Cancer Genome Atlas (TCGA). Data from the TMA and TCGA were analyzed for association of relapse-free survival (RFS) and overall survival (OS) with protein and gene expression of the chosen biomarkers.ResultsIncreased pAMPK protein activity by IHC and AMPK gene expression by TCGA gene expression data was correlated with improved RFS with a trend towards statistical significance.ConclusionsThis data suggests that increased pAMPK activity and expression may portend a better prognosis in HPV-associated HNSCC undergoing primary definitive resection. However, these findings require validation in larger studies.

Transient Receptor Potential (TRP) Channels in Head-and-Neck Squamous Cell Carcinomas: Diagnostic, Prognostic, and Therapeutic Potentials.

Head-and-neck squamous cell carcinomas (HNSCC) remain a leading cause of cancer morbidity and mortality worldwide. This is a largely preventable disease with smoking, alcohol abuse, and human papilloma virus (HPV) being the main risk factors. Yet, many patients are diagnosed with advanced disease, and no survival improvement has been seen for oral SCC in the past decade. Clearly, new diagnostic and prognostic markers are needed for early diagnosis and to guide therapy. Gene expression studies implied the involvement of transient receptor potential (TRP) channels in the pathogenesis of HNSCC. TRPs are expressed in normal epithelium where they play a key role in proliferation and differentiation. There is increasing evidence that the expression of TRP channels may change in HNSCC with important implications for diagnosis, prognosis, and therapy. In this review, we propose that TRP channel expression may afford a novel opportunity for early diagnosis of HNSCC and targeted molecular treatment.

Abstract 3711: Regulation of oncogenic targets by miR-99a-3p (the passenger strand of the miR-99a-duplex) in head and neck squamous cell carcinoma

Abstract Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer, with approximately 650,000 new cases diagnosed annually and 400,000 HNSCC-related deaths worldwide each year. HNSCC is typically diagnosed when already at an advanced stage. Despite advancements in surgery, radiation therapy, and chemotherapy, patients with advanced HNSCC have a poor prognosis. The median overall survival time for patients with recurrence and metastasis is 10-13 months in the setting of first-line chemotherapy and 6 months in the second-line setting. Many studies have shown that noncoding RNAs encoded by the human genome are functional and play critical roles in various cellular processes, e.g., cell growth, migration, invasion, and apoptosis. Among noncoding RNAs, microRNAs (miRNAs) are endogenous single-stranded RNA molecules comprising 19-22 nucleotides that function as fine-tuners of RNA expression. A single miRNA regulates many RNA transcripts, and bioinformatics studies have shown that more than half of protein-coding genes are controlled by miRNAs. Aberrantly expressed miRNAs are closely associated with cancer pathogenesis via the disruption of RNA networks within cancer cells. To identify novel oncogenic targets in head and neck squamous cell carcinoma (HNSCC), we have analyzed antitumor microRNAs (miRNAs) and their controlled molecular networks in HNSCC cells. Based on our miRNA signature in HNSCC, both strands of the miR-99a-duplex (miR-99a-5p: the guide strand, and miR-99a-3p: the passenger strand) are downregulated in cancer tissues. Moreover, low expression of miR-99a-5p and miR-99a-3p significantly predicts poor prognosis in HNSCC, and these miRNAs regulate cancer cell migration and invasion. We previously showed that passenger strands of miRNAs have antitumor functions. Here, we screened miR-99a-3p-controlled oncogenes involved in HNSCC pathogenesis. Thirty-two genes were identified as miR-99a-3p-regulated genes, and 10 genes (STAMBP, TIMP4, TMEM14C, CANX, SUV420H1, HSP90B1, PDIA3, MTHFD2, BCAT1, and SLC22A15) significantly predicted 5-year overall survival. Notably, among these genes, STAMBP, TIMP4, TMEM14C, CANX, and SUV420H1 were independent prognostic markers of HNSCC by multivariate analyses. We further investigated the oncogenic function of STAMBP in HNSCC cells using knockdown assays. Our data demonstrated that the aggressiveness of phenotypes in HNSCC cells was attenuated by siSTAMBP transfection. Moreover, aberrant STAMBP expression was detected in HNSCC clinical specimens by immunochemistry. Our strategy, i.e., identification of antitumor miRNAs and their targets, maybe an attractive tool to reveal novel prognostic and therapeutic targets in HNSCC. Citation Format: Reona Okada, Keiichi Koshizuka, Yasutaka Yamada, Shogo Moriya, Naoko Kikkawa, Takashi Kinoshita, Toyoyuki Hanazawa, Naohiko Seki. Regulation of oncogenic targets by miR-99a-3p (the passenger strand of the miR-99a-duplex) in head and neck squamous cell carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3711.

Abstract 5118: Proteogenomics characterization of HPV-negative head and neck squamous cell carcinomas

Abstract Patients with head and neck squamous cell carcinomas (HNSCCs) are treated with surgery, radiation, chemotherapy, and limited targeted therapies. Compared to human papillomavirus (HPV)-positive HNSCCs, HPV-negative cases have worse treatment response and prognosis and represent an unmet clinical need. We performed comprehensive proteogenomic characterization of tumor specimens, matched normal adjacent tissues (NATs), and blood samples from 109 HPV-negative HNSCC patients. This cohort is dominated by tumors from oral cavity (45, 41%) and larynx (49, 45%). Somatic mutation and somatic copy number analyses validated previously reported genomic aberrations in HPV-negative HNSCC. Proteomics analysis linked p53 loss of heterozygosity to increased expression of EPCAM, a stemness marker. Additionally, FAT1 truncation mutations were associated with increased expression of proteins involved in keratinization, a key feature of SCC differentiation. Deletions of 3p and 9p led to the loss of genes encoding p16, chemokine receptors, and interferon/JAK/STAT signaling pathway proteins, whereas amplifications of 3q and 11q led to overexpression of proteins involved in cell proliferation and anti-apoptosis pathways. Comparative analysis of tumor and NAT proteomes and phosphoproteomes identified putative diagnostic biomarkers and druggable targets, and proteogenomic integration further identified putative neoantigens. Tumor site-specific characterization associated epigenetic silencing of neurofilaments with laryngeal but not oral cavity SCC. Protein targets of FDA approved or investigational drugs for HNSCC treatment showed high inter-tumor heterogeneity in their protein abundances. DNA copy number and RNA expression level were good surrogates of protein abundance for some targets, such as EGFR and PD-L1, but they failed to reflect protein levels or kinase activities for other targets, such as MMP9 and MTOR. Thus, there is a critical need for protein biomarker-driven treatment stratification. Deconvolution of bulk tumor gene expression data revealed an immune-hot subgroup and an immune-cold subgroup. Immune-hot tumors broadly overexpressed multiple immune checkpoints including PD-L1, IDO1, and CTLA4, underscoring the necessity of combination immune checkpoint inhibition to improve treatment efficacy. Immune-cold tumors were characterized by smoking, chromosomal instability, and activation of the CDK4/6-pRb axis, suggesting they could be targeted by CDK4/6 inhibitors. We also noted that EGFR-amplified tumors frequently harbor copy number aberrations of downstream signaling components of the EGFR pathway. This may explain the low response rate of EGFR-amplified tumors to EGFR inhibitors, and targeting multiple pathway components, including EGFR, PIK3CA and STAT3, may be required for these tumors. In summary, our integrative proteogenomic characterization revealed multiple novel insights into the pathogenesis and treatment of HPV-negative HNSCCs. Citation Format: Chen Huang, Lijun Chen, Yize Li, Sara Savage, Michael Schnaubelt, Felipe V. Leprevost, Marcin Cieslik, Yongchao Dou, Bo Wen, Jonathan T. Lei, Kai Li, Eric Jaehnig, Zhiao Shi, Meenakshi Anurag, Jianbo Pan, Yingwei Hu, Rodrigo V. Eguez, David J. Clark, Matthew Wyczalkowski, Saravana M. Dhanasekaran, Chandan Kumar, Antonio Colaprico, Karsten Krug, Michael Gillette, D. R. Mani, Seungyeul Yoo, Jiayi Ji, Xiaoyu Song, Weiping Ma, Xi Steven Chen, Alex Pico, Nathan J. Edwards, Scott D. Jewell, Mathangi Thiagarajan, Emily S. Boja, Henry Rodriguez, Andrew Sikora, Pei Wang, Matthew Ellis, Gilbert S. Omenn, Li Ding, Alexey I. Nesvizhskii, Adel K. EI-Naggar, Daniel W. Chan, Hui Zhang, Bing Zhang, Clinical Proteomic Tumor Analysis Consortium. Proteogenomics characterization of HPV-negative head and neck squamous cell carcinomas [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5118.

Effects of Porphyromonas gingivalis and Fusobacterium nucleatum on inflammasomes and their regulators in H400 cells.

Inflammasomes are multiprotein complexes that regulate immune processes in response to infections and tissue damage. They modulate Interleukin-1beta (IL-1β) expression, a major proinflammatory cytokine. The inflammasome/IL-1β pathway is involved in head and neck squamous cell carcinoma (HNSCC) progression and the periodontal pathogens Fusobacterium nucleatum (Fn) and Porphyromonas gingivalis (Pg) have been reported to cause chronic inflammation in HNSCC. The aim of this study was to characterise the role of these pathogens in regulating inflammasome activity and the IL-1β response in HNSCC in vitro. An HNSCC cell line (H400) was exposed to Fn and Pg individually or in combination for 24h, ± incubation for 30min with 5mM adenosine triphosphate (ATP). Transcript levels of inflammasomes, NLRP3 and AIM2; inflammasome-regulatory proteins, POP1, CARD16 and TRIM16; and inflammasome-component, ASC and caspase 1 and IL-1β, were assayed by RT-PCR. Expression of IL-1β was by immunocytochemistry and ELISA. NLRP3 expression was significantly upregulated in response to Pg, Fn+Pg, Pg+ATP and Fn+Pg+ATP. AIM2 was significantly upregulated by Fn, Pg and Fn+Pg+ATP exposure. All conditions significantly upregulated IL-1β gene expression. POP1 expression was significantly downregulated by Pg or Fn exposure but not by Fn+Pg. Intracellular pro- and mature IL-1β were significantly higher following Fn and Pg+ATP exposure. Pg alone increased IL-1β by upregulating AIM2, NLRP3 and downregulating POP1. Fn promoted IL-1β by increasing AIM2 and downregulating POP1. Pg+ATP with or without Fn upregulated NLRP3, IL-1β by downregulating POP1. Periodontal pathogens may contribute to HNSCC pathogenesis by increasing the IL-1β response due to inflammasome dysregulation.

Abstract A31: Deregulation of NOTCH 1/NR4A2 signaling axis in head and neck cancer pathogenesis

Abstract Background: Notch1 and its intracellular cytoplasmic domain (NICD) are dysregulated in many solid tumors, including head and neck squamous cell carcinoma (HNSCC), and associated with disease initiation and progression. NICD-mediated upregulation of nuclear receptor related 1 (NURR1/NR4A2) is linked to differentiation/development, homeostasis, and cellular metabolism. However, the role and molecular mechanism(s) of Notch1/NICD- NR4A2 axis in HNSCC pathogenesis are still unknown and need to be investigated. Methods: The clinicopathologic importance of Notch1 and NR4A2 was investigated using HNSCC patient samples and publicly available databases. NR4A2 was stably knocked out (KO) and Notch1 signaling was blocked using small-molecule inhibitor (PF3084014) in HNSCC cell lines and analyzed for tumorigenic and metastatic potential. We also examined the whole-genome NR4A2 target genes using ChIP sequencing. Results: Both Notch1 and NR4A2 were highly overexpressed in HNSCC patients. Inhibition of Notch1 signaling or NR4A2 KO significantly decreased the clonogenicity and migratory potential of HNSCC cells. Our ChIP analysis revealed that NR4A2 regulates genes involved in metabolism, hypoxia, cell cycle progression, integrin and Wnt signaling. Interestingly, we observed that NR4A2 regulates stem cell markers including Sox2, Nanog, and CD44 and their expression was drastically reduced upon Notch 1 inhibition. Conclusion: Targeting Notch1-NR4A2 signaling axis may be crucial for inhibition of cancer stem cell-mediated HNSCC pathogenesis. Citation Format: Sanjib Chaudhary, Ramesh Pothuraju, Zafar Sayed, Dwight T. Jones, Surinder K. Batra, Muzafar A. Macha. Deregulation of NOTCH 1/NR4A2 signaling axis in head and neck cancer pathogenesis [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Optimizing Survival and Quality of Life through Basic, Clinical, and Translational Research; 2019 Apr 29-30; Austin, TX. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(12_Suppl_2):Abstract nr A31.

MiR-501-5p acts as an energetic regulator in head and neck squamous cell carcinoma cells growth and aggressiveness via reducing CLCA4.

Recent research have proved that miR-501-5p acted as a potent tumor biomarker in several cancers, excluding head and neck squamous cell carcinoma (HNSCC). The study intends to discover the potential function and mechanism of miR-501-5p in HNSCC. Data from TCGA database and qRT-PCR estimated the expression of miR-501-5p and Calcium activated Chloride Channel A4 (CLCA4). Cell proliferation, clone formation and transwell assays were performed to explore HNSCC cells biological behaviors. Luciferase assay was carried out to identify the interaction between miR-501-5p and CLCA4. miR-501-5p was profoundly up-regulated in HNSCC samples and promoted cells proliferation and metastasis. CLCA4, as a target of miR-501-5p, was connected with worse outcomes in HNSCC patients. Co-transfection assay proved that miR-501-5p/CLCA4 functioned as crucial regulators to affect HNSCC cells biological behaviors. Our study illustrated that miR-501-5p exhibited a tumor-promoting role on HNSCC by targeting CLCA4, providing a new insight for revealing the pathogenesis and treatment of HNSCC.

EXPRESSION OF TETRASPANINS CD9 AND CD63 IN HEAD AND NECK SQUAMOUS CELL CARCINOMA: AN IMMUNOHISTOCHEMICAL STUDY

Objective Tetraspanins are proteins involved in several cellular processes, including association with tumor progression. In this study, the immunoexpression of tetraspanins in head and neck squamous cell carcinoma (HNSCC) were assessed, aiming to evaluate the participation of these molecules in carcinogenesis. Study Design An immunohistochemical study was carried out using the tissue microarray technique to evaluate the tetraspanins CD9 and CD63. One hundred and twenty-one HNSCCs were analyzed (male, n = 97; female, n = 24; mean age, 60.4 years). The anatomic sites were 84 oral cavity, 23 oropharynx, 3 skin, 7 paranasal sinuses, 2 larynx, 1 nasopharynx, and 1 pharynx. Results Only 103 HNSCCs were available for analysis. For CD9, 69 (67%) HNSCCs were positive (46 oral cavity, 17 oropharynx, 5 paranasal sinuses, and 1 skin). For CD63, 11 (10.7%) HNSCCs were positive (7 oral cavity, 2 oropharynx, 1 paranasal sinuses, and 1 skin). Conclusions Our results suggest participation of tetraspanins CD9 and CD63 in the HNSCC pathogenesis. Further studies are necessary to determinate their prognostic effect with therapeutic implications. FAPESP grant 2016/11419-0. Tetraspanins are proteins involved in several cellular processes, including association with tumor progression. In this study, the immunoexpression of tetraspanins in head and neck squamous cell carcinoma (HNSCC) were assessed, aiming to evaluate the participation of these molecules in carcinogenesis. An immunohistochemical study was carried out using the tissue microarray technique to evaluate the tetraspanins CD9 and CD63. One hundred and twenty-one HNSCCs were analyzed (male, n = 97; female, n = 24; mean age, 60.4 years). The anatomic sites were 84 oral cavity, 23 oropharynx, 3 skin, 7 paranasal sinuses, 2 larynx, 1 nasopharynx, and 1 pharynx. Only 103 HNSCCs were available for analysis. For CD9, 69 (67%) HNSCCs were positive (46 oral cavity, 17 oropharynx, 5 paranasal sinuses, and 1 skin). For CD63, 11 (10.7%) HNSCCs were positive (7 oral cavity, 2 oropharynx, 1 paranasal sinuses, and 1 skin). Our results suggest participation of tetraspanins CD9 and CD63 in the HNSCC pathogenesis. Further studies are necessary to determinate their prognostic effect with therapeutic implications. FAPESP grant 2016/11419-0.

Regulation of Oncogenic Targets by miR-99a-3p (Passenger Strand of miR-99a-Duplex) in Head and Neck Squamous Cell Carcinoma.

To identify novel oncogenic targets in head and neck squamous cell carcinoma (HNSCC), we have analyzed antitumor microRNAs (miRNAs) and their controlled molecular networks in HNSCC cells. Based on our miRNA signature in HNSCC, both strands of the miR-99a-duplex (miR-99a-5p: the guide strand, and miR-99a-3p: the passenger strand) are downregulated in cancer tissues. Moreover, low expression of miR-99a-5p and miR-99a-3p significantly predicts poor prognosis in HNSCC, and these miRNAs regulate cancer cell migration and invasion. We previously showed that passenger strands of miRNAs have antitumor functions. Here, we screened miR-99a-3p-controlled oncogenes involved in HNSCC pathogenesis. Thirty-two genes were identified as miR-99a-3p-regulated genes, and 10 genes (STAMBP, TIMP4, TMEM14C, CANX, SUV420H1, HSP90B1, PDIA3, MTHFD2, BCAT1, and SLC22A15) significantly predicted 5-year overall survival. Notably, among these genes, STAMBP, TIMP4, TMEM14C, CANX, and SUV420H1 were independent prognostic markers of HNSCC by multivariate analyses. We further investigated the oncogenic function of STAMBP in HNSCC cells using knockdown assays. Our data demonstrated that the aggressiveness of phenotypes in HNSCC cells was attenuated by siSTAMBP transfection. Moreover, aberrant STAMBP expression was detected in HNSCC clinical specimens by immunohistochemistry. This strategy may contribute to the clarification of the molecular pathogenesis of this disease.

HBD3 Induces PD-L1 Expression on Head and Neck Squamous Cell Carcinoma Cell Lines.

Human β-defensin 3 (HBD3) is an antimicrobial peptide up-regulated in the oral tissues of individuals with head and neck squamous cell carcinomas (HNSCC) and oral squamous cell carcinomas (SCC) and present in high concentrations in their saliva. In this study, we determined if HBD3 contributes to HNSCC pathogenesis by inducing programmed death-ligand 1 (PD-L1) expression on HNSCC cell lines. For this, SCC cell lines SCC4, SCC15, SCC19, SCC25, and SCC99 (5.0 × 104 viable cells) were used. Cells were incubated with IFNγ (0.6 µM) and HBD3 (0.2, 2.0, or 20.0 µM) for 24 h. Cells alone served as controls. Cells were then treated with anti-human APC-CD274 (PD-L1) and Live/Dead Fixable Green Dead Cell Stain. Cells treated with an isotype antibody and cells alone served as controls. All cell suspensions were analyzed in a LSR II Violet Flow Cytometer. Cytometric data was analyzed using FlowJo software. Treatment with IFNγ (0.6 µM) increased the number of cells expressing PD-L1 (p < 0.05) with respect to controls. Treatment with HBD3 (20.0 µM) also increased the number of cells expressing PD-L1 (p < 0.05) with respect to controls. However, treatment with IFNγ (0.6 µM) was not significantly different from treatment with HBD3 (20.0 µM) and the numbers of cells expressing PD-L1 were similar (p = 1). Thus, HBD3 increases the number of cells expressing PD-L1. This is a novel concept, but the role HBD3 contributes to HNSCC pathogenesis by inducing PD-L1 expression in tumors will have to be determined.

Somatic mutations and promotor methylation of the ryanodine receptor 2 is a common event in the pathogenesis of head and neck cancer.

Genomic sequencing projects unraveled the mutational landscape of head and neck squamous cell carcinoma (HNSCC) and provided a comprehensive catalog of somatic mutations. However, the limited number of significant cancer-related genes obtained so far only partially explains the biological complexity of HNSCC and hampers the development of novel diagnostic biomarkers and therapeutic targets. We pursued a multiscale omics approach based on whole-exome sequencing, global DNA methylation and gene expression profiling data derived from tumor samples of the HIPO-HNC cohort (n = 87), and confirmed new findings with datasets from The Cancer Genome Atlas (TCGA). Promoter methylation was confirmed by MassARRAY analysis and protein expression was assessed by immunohistochemistry and immunofluorescence staining. We discovered a set of cancer-related genes with frequent somatic mutations and high frequency of promoter methylation. This included the ryanodine receptor 2 (RYR2), which showed variable promoter methylation and expression in both tumor samples and cell lines. Immunohistochemical staining of tissue sections unraveled a gradual loss of RYR2 expression from normal mucosa via dysplastic lesion to invasive cancer and indicated that reduced RYR2 expression in adjacent tissue and precancerous lesions might serve as risk factor for unfavorable prognosis and upcoming malignant conversion. In summary, our data indicate that impaired RYR2 function by either somatic mutation or epigenetic silencing is a common event in HNSCC pathogenesis. Detection of RYR2 expression and/or promoter methylation might enable risk assessment for malignant conversion of dysplastic lesions.

Rola układu immunologicznego w patogenezie oraz progresji płaskonabłonkowego raka głowy i szyi

Płaskonabłonkowy rak głowy i szyi (HNSCC, ang. Head and Neck Squamous Cell Carcinoma) to nowotwór wywodzący się z komórek płaskonabłonkowych błon śluzowych jamy ustnej, nosa lub gardła. Pomimo znacznych postępów konwencjonalnych metod leczenia, 5-letni wskaźnik przeżycia bez progresji u pacjentów z miejscowo zaawansowanym HNSCC HPV(-) nadal nie przekracza 50%. Niskie wskaźniki przeżycia w połączeniu z wysoką toksycznością stosowanych terapii, podkreślają konieczność wprowadzenia nowych strategii terapeutycznych. Obecne badania potwierdzają, że układ immunologiczny odgrywa kluczową rolę w patogenezie HNSCC. Lepsze zrozumienie mechanizmów molekularnych odpowiedzialnych za ucieczkę komórek nowotworowych spod kontroli układu immunologicznego oraz dokładne poznanie szlaków molekularnych odpowiedzialnych za rozwój płaskonabłonkowego raka głowy i szyi umożliwi opracowanie bardziej efektywnych terapii. Celem pracy jest przegląd wiedzy dotyczącej roli układu odpornościowego w ucieczce komórek nowotworowych HNSCC spod nadzoru immunologicznego.

HOTTIP Functions as a Key Candidate Biomarker in Head and Neck Squamous Cell Carcinoma by Integrated Bioinformatic Analysis.

Background Accumulating evidence has demonstrated the pivotal role of long noncoding RNAs (lncRNAs) in competing endogenous RNA (ceRNA) networks for predicting survival and evaluating prognosis in cancer patients. However, the pathogenesis of head and neck squamous cell carcinoma (HNSCC) remains unclear, and prognostic biomarkers for HNSCC are still lacking. Methods A total of 546 RNA sequencing profiles of HNSCC patients with clinical outcome data were obtained from the Cancer Genome Atlas (TCGA) database, providing a large sample of RNA sequencing data. From these, 71 Long noncoding RNAs lncRNAs, 8 microRNAs (miRNAs), and 16 messenger RNAs (mRNAs) were identified to construct a HNSCC-specific ceRNA network (fold change >2, P < 0.05). Univariate and multivariate Cox proportional regression models were used to assess independent indicators of prognosis. Then the expression of lncRNAs harboring prognostic value was validated in human HNSCC cell lines and tumor samples from our cohort and another two datasets from GEO (Gene Expression Omnibus) databases. Results As a result, a 3-mRNA signature and 6-lncRNA signature were identified. The six-lncRNA signature exhibited the highest prognostic value. Notably, in the six lncRNAs, HOTTIP showed the greatest prognostic value and was significantly correlated with clinical stage and histological grade of HNSCC patients. Furthermore, it was proved that HOTTIP was upregulated in HNSCC cell lines and cancerous tissues compared with corresponding normal cell lines and normal tissues. Functional assessment analysis revealed that HOTTIP might play a key role in the oncogenesis and progression of HNSCC. Conclusion The present study deepened our understanding of the ceRNA-related regulatory mechanism in the pathogenesis of HNSCC and identified candidate prognostic biomarkers for clinical outcome prediction in HNSCC. HOTTIP may function as a key candidate biomarker in HNSCC and serve as a prognostic marker for HNSCC patients.

Impact of Epigenetic Regulation on Head and Neck Squamous Cell Carcinoma

The most common type of head and neck cancer, head and neck squamous cell carcinoma (HNSCC), can develop therapeutic resistance that complicates its treatment. The 5-y survival rate for HNSCC remains at ~50%, and improving these outcomes requires a better understanding of the pathogenesis of HNSCC. Studies of HNSCC using in vitro, ex vivo, and in vivo approaches provide a novel conceptual framework based on epigenetic mechanisms for developing future clinical applications. Normal oral tissues are influenced by environmental factors that induce pathological changes affecting the network of epigenetic enzymes and signaling pathways to induce HNSCC growth and metastasis. Although various epigenetic regulator families, such as DNA methyltransferases, ten-eleven translocation proteins, histone acetyltransferases, histone deacetylases, BET bromodomain proteins, protein arginine methyltransferases, histone lysine methyltransferases, and histone lysine demethylases, have a role in diverse cancers, specific members have a function in HNSCC. Recently, lysine-specific demethylases have been identified as a potential, attractive, and novel target of HNSCC. Lysine-specific demethylase 1 (LSD1) expression is inappropriately upregulated in HNSCC and an orthotopic HNSCC mouse model. LSD1 can demethylate lysine at specific histone positions to repress gene expression or stimulate transcription, indicating a dual and context-dependent role in transcriptional regulation. Our study showed that LSD1 promotes HNSCC growth and metastasis. Pharmacological attenuation of LSD1 inhibits orthotopic and patient-derived HNSCC xenograft growth-specific target genes and signaling pathways. This review provides recent evidence demonstrating the function of epigenetic regulator enzymes in HNSCC progression, including potential therapeutic applications for such enzymes in combination and immunotherapy.

Muscle-Specific FXR1 Isoforms in Squamous Cell Cancer

The RNA-binding protein fragile-X mental retardation autosomal 1 (FXR1) is upregulated in head and neck squamous cell carcinomas (HNSCCs) and expressed as at least seven isoforms in humans. Only two of these isoforms are capable of binding to RNA containing G-quadruplex structures. We suggest that these unique isoforms play a role in the pathogenesis of HNSCC.

A Review of HPV-Related Head and Neck Cancer.

Head and neck squamous cell carcinomas (HNSCCs) arise in the mucosal lining of the upper aerodigestive tract. Tobacco and alcohol use have been reported to be associated with HNSCC. Infection with high-risk human papillomaviruses (HPVs) has recently been implicated in the pathogenesis of HNSCCs. It is now widely accepted that high-risk HPV is a cause of almost all cervical cancers as well as some forms of HNSCCs. HPV-related HNSCCs are increasing. HPV-related HNSCCs and HPV-unrelated HNSCCs differ with respect to the molecular mechanisms underlying their oncogenic processes. HPV-related HNSCCs are known to have a better prognosis response to treatment as compared with HPV-unrelated HNSCCs. Therefore, in recent years, it has been required to accurately discriminate between HPV-related and HPV-unrelated HNSCCs. To diagnose the HPV-related HNSCCs, various methods including P16 immunohistochemistry, FISH, and genetic analyses of the HPV gene from histopathological and liquid biopsy specimens have been employed. Based on the results of the differential diagnosis, various treatments employing EGFR TKI and low-dose radiation have been employed. Here, we review the involvement of the HPV virus in HNSCCs as well as the molecular mechanism of carcinogenesis, classification, prognosis, diagnostic procedures, and therapy of the disease.