DNA methylation-based depiction of the immune microenvironment and immune-associated long non-coding RNAs in oral cavity squamous cell carcinomas.

Oral cavity squamous cell carcinoma (OSCC) is a complex and dynamic disease characterized by clinicopathological and molecular heterogeneity. Spatial and temporal heterogeneity of cell subpopulations has been associated with cancer progression and implicated in the prognosis and therapy response. Emerging evidence indicates that aberrant epigenetic profiles in OSCC may foster an immunosuppressive tumor microenvironment by modulating the expression of immune-related long non-coding RNAs (lncRNAs). DNA methylation analysis was performed in 46 matched OSCC and normal adjacent tissue samples using a genome-wide platform (Infinium HumanMethylation450 BeadChip). Reference-based computational deconvolution (MethylCIBERSORT) was applied to infer the immune cell composition of the bulk samples. The expression levels of genes encoding immune markers and differentially methylated lncRNAs were investigated using The Cancer Genome Atlas dataset. OSCC specimens presented distinct immune cell composition, including the enrichment of monocyte lineage cells, natural killer cells, cytotoxic T-lymphocytes, regulatory T-lymphocytes, and neutrophils. In contrast, B-lymphocytes, effector T-lymphocytes, and fibroblasts were diminished in tumor samples. The hypomethylation of three immune-associated lncRNAs (MEG3, MIR155HG, and WFDC21P) at individual CpG sites was confirmed by bisulfite-pyrosequencing. Also, the upregulation of a set of immune markers (FOXP3, GZMB, IL10, IL2RA, TGFB, IFNG, TDO2, IDO1, and HIF1A) was detected. The immune cell composition, immune markers alteration, and dysregulation of immune-associated lncRNAs reinforce the impact of the immune microenvironment in OSCC. These concurrent factors contribute to tumor heterogeneity, suggesting that epi-immunotherapy could be an efficient alternative to treat OSCC.

Decreased CD1a + and CD83 + cells in tonsillar squamous cell carcinoma regardless of HPV status.

BACKGROUND: Dendritic cells (DCs) are specialized antigen-presenting cells that play a critical role in the immune response against human papillomavirus (HPV) infection, and represent a therapeutic target in cancer. OBJECTIVE: To identify and quantify DCs in tonsillar squamous cell carcinoma (TSCC) under the influence of HPV infection. METHODOLOGY: CD1a and CD83 antibodies were used to identify immature dendritic cells and mature dendritic cells by immunohistochemistry in 33 primary TSCC and 10 normal tonsils (NTs), respectively. For the TSCC samples, the number of DCs per area was evaluated in the intra- and peritumoral compartments. For the NTs, the quantification of DCs was evaluated in the intra- and peritonsillar compartments. HPV detection methods were determined according to the ASCO Clinical Practice Guidelines from the College of American Pathologists Guideline (2018). RESULTS: There were fewer intratumoral CD1a+ DCs in the HPV-positive and HPV-negative TSCC groups than in the NT group (p<0.05). In the peritumoral compartment, there were fewer CD83+ DCs in the HPV-positive and HPV-negative TSCC groups than in the NT group (p<0.001). The quantification of DCs subtypes showed no statistical differences between HPV-positive and HPV-negative TSCC groups (p>0.137). Patients with HPV-positive TSCC had significantly better overall survival rate than those with HPV-negative TSCC (p=0.004). CONCLUSION: Tumor activity contributes to DC depletion regardless of intralesional HPV positivity. An improved prognosis has been reported in patients with HPV-positive TSCC.

Decreased CD1a + and CD83 + cells in tonsillar squamous cell carcinoma regardless of HPV status

Abstract Dendritic cells (DCs) are specialized antigen-presenting cells that play a critical role in the immune response against human papillomavirus (HPV) infection, and represent a therapeutic target in cancer. Objective: To identify and quantify DCs in tonsillar squamous cell carcinoma (TSCC) under the influence of HPV infection. Methodology: CD1a and CD83 antibodies were used to identify immature dendritic cells and mature dendritic cells by immunohistochemistry in 33 primary TSCC and 10 normal tonsils (NTs), respectively. For the TSCC samples, the number of DCs per area was evaluated in the intra- and peritumoral compartments. For the NTs, the quantification of DCs was evaluated in the intra- and peritonsillar compartments. HPV detection methods were determined according to the ASCO Clinical Practice Guidelines from the College of American Pathologists Guideline (2018). Results: There were fewer intratumoral CD1a+ DCs in the HPV-positive and HPV-negative TSCC groups than in the NT group (p<0.05). In the peritumoral compartment, there were fewer CD83+ DCs in the HPV-positive and HPV-negative TSCC groups than in the NT group (p<0.001). The quantification of DCs subtypes showed no statistical differences between HPV-positive and HPV-negative TSCC groups (p>0.137). Patients with HPV-positive TSCC had significantly better overall survival rate than those with HPV-negative TSCC (p=0.004). Conclusion: Tumor activity contributes to DC depletion regardless of intralesional HPV positivity. An improved prognosis has been reported in patients with HPV-positive TSCC.

Impact of smoking on dendritic cells in patients with oral squamous cell carcinoma.

Smoking has been shown to alter innate and adaptive immune responses and is directly associated with the onset of oral squamous cell carcinoma (OSCC). The purpose of this study was to evaluate the effect of cigarette smoke exposure on dendritic cells (DCs) from OSCC patients. CD1a and CD83 antibodies were used to identify immature and mature DCs, respectively, by immunohistochemistry in OSCC samples of 24 smokers and 24 non-smokers. Density of DCs was calculated in intra and peritumoral areas. Clinical and microscopic findings were reviewed and analyzed for all patients. Smokers with OSCC had a lower density of intra and peritumoral DCs when compared to non-smokers. Tumors classified as moderately/poorly differentiated had lower peritumoral CD1a+ DCs than well-differentiated tumors (p < 0.001). Smoking contributed to a depletion of immature and mature DCs in the OSCC.

Impact of smoking on dendritic cells in patients with oral squamous cell carcinoma

Abstract Smoking has been shown to alter innate and adaptive immune responses and is directly associated with the onset of oral squamous cell carcinoma (OSCC). The purpose of this study was to evaluate the effect of cigarette smoke exposure on dendritic cells (DCs) from OSCC patients. CD1a and CD83 antibodies were used to identify immature and mature DCs, respectively, by immunohistochemistry in OSCC samples of 24 smokers and 24 non-smokers. Density of DCs was calculated in intra and peritumoral areas. Clinical and microscopic findings were reviewed and analyzed for all patients. Smokers with OSCC had a lower density of intra and peritumoral DCs when compared to non-smokers. Tumors classified as moderately/poorly differentiated had lower peritumoral CD1a+ DCs than well-differentiated tumors (p < 0.001). Smoking contributed to a depletion of immature and mature DCs in the OSCC.

Clinicopathologic analysis of 14 cases of odontogenic myxoma and review of the literature.

BACKGROUND: Odontogenic myxoma is a rare benign neoplasm that originates from odontogenic ectomesenchyme. There is no standard of care and recurrences are frequent after conservative surgical procedures. MATERIAL AND METHODS: A retrospective study conducted at a single cancer center, with analysis of medical records of all patients diagnosed with odontogenic myxoma from 1980 to 2010, along with a literature review. RESULTS: There were 14 patients with diagnosis of odontogenic myxoma (OM). Most patients were female (78.6%) and Caucasian (100%), with ages ranging from 7 to 51 years (21.6 ± 11.6 years). The time period between the first symptom and first consultation ranged from 0 to 60 months (19.4 ± 19.97 months). The most frequent complaints were increased local volume or failure to tooth eruption. The most common tumor site was the mandible (11 cases, 78.5%). About radiological findings, most lesions were multilocular (9 cases, 64.3%) and with imprecise limits (12 cases, 85.7%). Surgery was performed in all cases and curettage was the most applied technique (10 cases, 71.4%). Three patients underwent mandibulectomy and complex reconstructions including iliac crest microvascular flap. Three patients had postoperative complications and 4 had local recurrences of the tumor. The follow up time ranged from 12 to 216 months (112 ± 70.8 months). All patients are without clinical and radiographic evidence of disease. CONCLUSIONS: OM is a locally aggressive and rare tumor. There is no gold standard surgical management and the therapeutic decision should be individualized taking into account the characteristics and extension of the tumor. Key words:Mandible, myxoma, odontogenic, odontogenic tumor.

Correlation between swallowing-related quality of life and videofluoroscopy after head and neck cancer treatment. / Associação entre os achados do questionário de disfagia M. D. Anderson e a videofluoroscopia da deglutição após tratamento do câncer de cabeça e pescoço.

INTRODUCTION: The use of symptom-specific questionnaires on head and neck cancer (HNC), together with objective swallowing measures, can be sensitive to changes in quality of life (QoL) resulting from dysphagia, but this tool is not broadly used as a complement to clinical evaluations. PURPOSE: To analyze the correlation between the M. D. Anderson Dysphagia Inventory (MDADI) questionnaire and videofluoroscopy (VF) in patients treated for head and neck cancer. METHODS: This is a retrospective study with review of clinical data, VF and MDADI results. The study sample was composed of adult patients (>18 y.o.) treated for tumors at the oral cavity, oropharynx, hypopharynx, and larynx, regardless of treatment type. For the VF examination, swallowing of 5 and 20 ml of nectar-thick liquids were considered. The Mann-Whitney nonparametric test was applied to evaluate the correlations between the MDADI and VF. RESULTS: Thirty-nine patients, mostly men (87.18%), with mean age of 61 years participated in the study. Most patients (16) presented oral cavity tumors (41.03%). Twenty-two patients were in advanced clinical stage (IV). Surgery was the most prevalent treatment (41.03%). Approximately half of the participants (20) received oral feeding. The total mean (TM) on the MDADI was 63.36. Comparison between VF and MDADI data showed significant correlation between TM, emotional domain (ED), and physical domain (PD) with penetration during the swallowing of 5 ml. Penetration and aspiration with 20 ml determined worse QoL on the global (p=0.018 and p=0.0053), emotional (p=0.0012 and p=0.027) and physical (p=0.0002 and p=0.0051) domains, and TM (p=0.0023 and p=0.0299), respectively. The presence of stasis did not determine worse QoL. CONCLUSION: Patients treated for HNC who presented penetration/aspiration showed worse QoL on the emotional and physical domains of the MDADI.

Associação entre os achados do questionário de disfagia M. D. Anderson e a videofluoroscopia da deglutição após tratamento do câncer de cabeça e pescoço / Correlation between swallowing-related quality of life and videofluoroscopy after head and neck cancer treatment

RESUMO Introdução A utilização de questionários sintoma-específicos no câncer de cabeça e pescoço (CCP) em conjunto com avaliações objetivas da deglutição pode ser sensível às mudanças na qualidade de vida (QV) decorrentes da disfagia, porém é uma ferramenta pouco utilizada como complemento de avaliações clínicas. Objetivo analisar a associação entre o questionário de disfagia M. D. Anderson (MDADI) com a videofluoroscopia (VF) da deglutição em pacientes tratados do CCP. Método Estudo retrospectivo, com revisão de prontuários, dados da VF e do questionário de disfagia MDADI. Foram incluídos indivíduos maiores de 18 anos, tratados do câncer de cavidade oral, orofaringe, hipofaringe e laringe, independentemente do tratamento curativo. Para o exame de VF, foram consideradas as deglutições de 5 e 20 ml na consistência néctar. O teste não paramétrico de Mann-Whitney foi utilizado para avaliar a associação entre o questionário MDADI e a VF. Resultados Casuística de 39 indivíduos, predomínio de homens, 34 (87,18%), e média de idade de 61 anos. Prevalência de câncer de cavidade oral, 16 (41,03%). Vinte e dois (56,4%) possuíam estádio clínico IV. Cirurgia isolada foi o tratamento mais prevalente, 16 (41,03%). Vinte indivíduos (51,28%) se alimentavam por via oral. A média total (MT) do MDADI foi de 63,36. Na correlação da VF com o MDADI, observou-se associação significante entre MT, domínio emocional (DE) e domínio físico (DFis) com penetração para 5 ml. Penetração e aspiração com 20 ml determinou prejuízo para questão global (p=0,018 e p=0,0053), DE (p=0,0012 e p=0,027), DFis (p=0,0002 e p=0,0051) e MT (p=0,0023 e p=0,0299), respectivamente. A presença de estase não determinou piora da QV. Conclusão Pacientes tratados do CCP que apresentam penetração/aspiração demonstram impacto na qualidade de vida nos DE e DFis.

ABSTRACT Introduction The use of symptom-specific questionnaires on head and neck cancer (HNC), together with objective swallowing measures, can be sensitive to changes in quality of life (QoL) resulting from dysphagia, but this tool is not broadly used as a complement to clinical evaluations. Purpose To analyze the correlation between the M. D. Anderson Dysphagia Inventory (MDADI) questionnaire and videofluoroscopy (VF) in patients treated for head and neck cancer. Methods This is a retrospective study with review of clinical data, VF and MDADI results. The study sample was composed of adult patients (>18 y.o.) treated for tumors at the oral cavity, oropharynx, hypopharynx, and larynx, regardless of treatment type. For the VF examination, swallowing of 5 and 20 ml of nectar-thick liquids were considered. The Mann-Whitney nonparametric test was applied to evaluate the correlations between the MDADI and VF. Results Thirty-nine patients, mostly men (87.18%), with mean age of 61 years participated in the study. Most patients (16) presented oral cavity tumors (41.03%). Twenty-two patients were in advanced clinical stage (IV). Surgery was the most prevalent treatment (41.03%). Approximately half of the participants (20) received oral feeding. The total mean (TM) on the MDADI was 63.36. Comparison between VF and MDADI data showed significant correlation between TM, emotional domain (ED), and physical domain (PD) with penetration during the swallowing of 5 ml. Penetration and aspiration with 20 ml determined worse QoL on the global (p=0.018 and p=0.0053), emotional (p=0.0012 and p=0.027) and physical (p=0.0002 and p=0.0051) domains, and TM (p=0.0023 and p=0.0299), respectively. The presence of stasis did not determine worse QoL. Conclusion Patients treated for HNC who presented penetration/aspiration showed worse QoL on the emotional and physical domains of the MDADI.

Alcohol and tobacco consumption affects bacterial richness in oral cavity mucosa biofilms.

BACKGROUND: Today there are more than 2 billion alcohol users and about 1.3 billion tobacco users worldwide. The chronic and heavy use of these two substances is at the heart of numerous diseases and may wreak havoc on the human oral microbiome. This study delves into the changes that alcohol and tobacco may cause on biofilms of the human oral microbiome. To do so, we used swabs to sample the oral biofilm of 22 subjects; including 9 control-individuals with no or very low consumption of alcohol and no consumption of tobacco, 7 who were chronic and heavy users of both substances and 6 active smokers that reported no significant alcohol consumption. DNA was extracted from swabs and the V1 region of the 16S rRNA gene was PCR amplified and sequenced using the Ion Torrent PGM platform, generating 3.7 million high quality reads. DNA sequences were clustered and OTUs were assigned using the ARB SILVA database and Qiime. RESULTS: We found no differences in species diversity and evenness among the groups. However, we found a significant decrease in species richness in only smokers and in smokers/drinkers when compared to controls. We found that Neisseria abundance was significantly decreased in both groups when compared to controls. Smokers had significant increases in Prevotella and Capnocytophaga and reductions in Granulicatella, Staphylococcus, Peptostreptococcus and Gemella when compared to the two other groups. Controls showed higher abundance of Aggregibacter, whilst smokers/drinkers had lower abundances of Fusobacteria. Samples from only smokers clustered closer together than to controls and smokers/drinkers, and also had a significant reduction in inter-group dissimilarity distances, indicating a more homogenous group than controls. CONCLUSIONS: Our results indicate that the continued use of tobacco or alcohol plus tobacco significantly reduces bacterial richness, which apparently leads to a reduction in inter-group variability, turning the respective biofilms into a more homogenous microenvironment in terms of bacterial community composition, with possible consequences for human oral diseases.

Fluorescence spectroscopy for the detection of potentially malignant disorders and squamous cell carcinoma of the oral cavity.

Oral cancer is a public health problem with relevant incidence in the world population. The affected patient usually presents advanced stage disease and the consequence of this delay is a reduction in survival rates. Given this, it is essential to detect oral cancer at early stages. Fluorescence spectroscopy is a non-invasive diagnostic tool that can improve cancer detection in real time. It is a fast and accurate technique, relatively simple, which evaluates the biochemical composition and structure using the tissue fluorescence spectrum as interrogation data. Several studies have positive data regarding the tools for differentiating between normal mucosa and cancer, but the difference between cancer and potentially malignant disorders is not clear. The aim of this study was to evaluate the efficacy of fluorescence spectroscopy in the discrimination of normal oral mucosa, oral cancer, and potentially malignant disorders. The fluorescence spectroscopy was evaluated in 115 individuals, of whom 55 patients presented oral squamous cell carcinoma, 30 volunteers showing normal oral mucosa, and 30 patients having potentially malignant disorders. The spectra were classified and compared to histopathology to evaluate the efficiency in diagnostic discrimination employing fluorescence. In order to classify the spectra, a decision tree algorithm (C4.5) was applied. Despite of the high variance observed in spectral data, the specificity and sensitivity obtained were 93.8% and 88.5%, respectively at 406 nm excitation. These results point to the potential use of fluorescence spectroscopy as an important tool for oral cancer diagnosis and potentially malignant disorders.

Analysis of surgical margins in oral cancer using in situ fluorescence spectroscopy.

UNLABELLED: Oral cancer is a public health problem with high prevalence in the population. Local tumor control is best achieved by complete surgical resection with adequate margins. A disease-free surgical margin correlates with a lower rate of local recurrence and a higher rate of disease-free survival. Fluorescence spectroscopy is a noninvasive diagnostic tool that can aid in real-time cancer detection. The technique, which evaluates the biochemical composition and structure of tissue fluorescence, is relatively simple, fast and, accurate. OBJECTIVES: This study aimed to compare oral squamous cell carcinoma lesions to surgical margins and the mucosa of healthy volunteers by fluorescence spectroscopy. MATERIALS AND METHODS: The sample consisted of 56 individuals, 28 with oral squamous cell carcinoma and 28 healthy volunteers with normal oral mucosa. Thirty six cases (64.3%) were male and the mean age was 60.9 years old. The spectra were classified and compared to histopathology to determine fluorescence efficiency for diagnostic discrimination of tumors. RESULTS: In the analysis of the other cases we observed discrimination between normal mucosa, injury and margins. At two-year follow up, three individuals had local recurrence, and in two cases investigation fluorescence in the corresponding area showed qualitative differences in spectra between the recurrence area and the area without recurrence at the same anatomical site in the same patient. CONCLUSION: In situ analysis of oral mucosa showed the potential of fluorescence spectroscopy as a diagnostic tool that can aid in discrimination of altered mucosa and normal mucosa.