A biomimetic nanoplatform for customized photothermal therapy of HNSCC evaluated on patient-derived xenograft models / 国际口腔科学杂志·英文版

Cancer cell membrane (CCM) derived nanotechnology functionalizes nanoparticles (NPs) to recognize homologous cells, exhibiting translational potential in accurate tumor therapy. However, these nanoplatforms are majorly generated from fixed cell lines and are typically evaluated in cell line-derived subcutaneous-xenografts (CDX), ignoring the tumor heterogeneity and differentiation from inter- and intra- individuals and microenvironments between heterotopic- and orthotopic-tumors, limiting the therapeutic efficiency of such nanoplatforms. Herein, various biomimetic nanoplatforms (CCM-modified gold@Carbon, i.e., Au@C-CCM) were fabricated by coating CCMs of head and neck squamous cell carcinoma (HNSCC) cell lines and patient-derived cells on the surface of Au@C NP. The generated Au@C-CCMs were evaluated on corresponding CDX, tongue orthotopic xenograft (TOX), immune-competent primary and distant tumor models, and patient-derived xenograft (PDX) models. The Au@C-CCM generates a photothermal conversion efficiency up to 44.2% for primary HNSCC therapy and induced immunotherapy to inhibit metastasis via photothermal therapy-induced immunogenic cell death. The homologous CCM endowed the nanoplatforms with optimal targeting properties for the highest therapeutic efficiency, far above those with mismatched CCMs, resulting in distinct tumor ablation and tumor growth inhibition in all four models. This work reinforces the feasibility of biomimetic NPs combining modular designed CMs and functional cores for customized treatment of HNSCC, can be further extended to other malignant tumors therapy.

Avaliação in vitro e in silico da atividade do S-(-)-Álcool Perílico sobre linhagens celulares do Carcinoma Epidermóide de Língua / In vitro and in silico evaluation of the activity of S-(-)-Peryllic Alcohol on Tongue Squamous Carcinoma cell lines

O carcinoma epidermóide oral (CEO) é a neoplasia maligna mais frequente da cavidade oral e constitui um problema de saúde pública devido a sua alta taxa de incidência e mortalidade devido em muitos casos ao fracasso terapêutico e a resistência tumoral. Assim sendo, destaca-se a busca por novas moléculas biologicamente ativas, como as encontradas nos produtos de origem natural. Este trabalho tem como objetivo avaliar a atividade antineoplásica do S-(-)-álcool perílico (POH) em culturas de células de CEO de língua e predizer sua afinidade através de modelo computacional sobre proteínas que regulam o ciclo celular. Para isso, foram utilizadas duas linhagens celulares de CEO de língua, HSC-3 e SCC-25. Os seguintes grupos foram analisados: G0 (controle; células cultivadas na ausência de POH), G1 (células tratadas com cisplatina a 40 µM), G2 (células tratadas com POH a 0,5 mM), G3 (células tratadas com POH a 1,0 mM), G4 (células tratadas com POH a 1,5 mM) e G5 (células tratadas com POH a 3,0 mM). Diferenças entre estes grupos foram investigadas através dos seguintes ensaios: viabilidade celular (Alamar Blue e Live/Dead assay) e atividade migratória (Wound healing). Foi também realizada a predição de afinidade entre o POH e as moléculas de controle do ciclo celular utilizando a docagem molecular com emprego do software Molegro Virtual Docker, v. 6.0.1. Os dados foram tratados estatisticamente pelo GraphPad Prism 6.0 (GraphPad Software, EUA), análises paramétricas utilizando teste Anova, pós-teste de Tukey e teste estatístico não-paramétricos de Kruskal-Wallis, seguido pelo teste t de estudent foram adotados para determinação de diferenças entre os grupos experimentais. O índice de significância considerado neste trabalho foi de 5%. Para ambas as técnicas de avaliação da viabilidade celular (Alamar Blue e Live/dead assay) analisadas neste trabalho, o POH foi capaz de reduzir a viabilidade celular de linhagens do CEO de língua de maneira dosedependente e tempo-dependente (p<0,05). As concentrações de 1,5 mM e 3 mM do POH obtiveram resultados melhores ou semelhantes aos encontrados na cisplatina 40 µM, para as duas linhagens, na avaliação da viabilidade celular (p<0,05). Os valores de IC50 do POH foram de 1,5 mM para a célula SCC-25 em todos os intervalos de tempo (24 h, 48 h e 72 h), uma vez que, para a linhagem HSC-3, foram de 3 mM para os tempos de 24 h e 48 h e de 1,5 mM para o intervalo de 72 h. O POH foi capaz de inibir a migração das duas linhagens celulares de CEO de maneira dependente da concentração (p≤0,05), comparados ao grupo controle. A habilidade da molécula POH se ligar a proteínas responsáveis pela ativação do ciclo celular foi avaliada usando docking models. Dentre elas, a proteína GTPase Kras mostrou a melhor energia de ligação (-86.70 kcal/mol), apresentando ligações de hidrogênio com os resíduos THR58 (A) e ASP57 (A) e ligações estéricas com os resíduos TRY32 (A) e ALA18 (A). As evidências deste estudo corroboram a ideia de que o POH possui atividade sobre o CEO, sugerindo que essa molécula possa ser uma forte candidata para o desenvolvimento de medicamentos direcionados ao tratamento desta patologia (AU).

Oral squamous cell carcinoma (OSCC) is the most frequent malignant neoplasm of the oral cavity and constitutes a public health problem due to its high incidence and mortality rate caused in many cases by therapeutic failure and tumor resistance. Therefore, the search for new biologically active molecules stands out, such as those found in products of natural origin. This work aims to evaluate the antineoplastic activity of S-(-)-perillyl alcohol (POH) in cell cultures of tongue CEO and to predict its affinity through a computer model on proteins that regulate the cell cycle. For this purpose, two cell lines of tongue CEO were used, HSC-3 and SCC-25. The following groups were analyzed: G0 (control; cells cultured in the absence of POH), G1 (cells treated with 40 µM cisplatin), G2 (cells treated with 0.5 mM POH), G3 (cells treated with 1 .0 mM), G4 (cells treated with 1.5 mM POH) and G5 (cells treated with 3.0 mM POH). Differences between these groups were investigated through the following assays: cell viability (Alamar Blue and Live/Dead assay) and migratory activity (Wound healing). Affinity prediction between POH and cell cycle control molecules were also performed using molecular docking using Molegro Virtual Docker, v. 6.0.1. The data was statistically treated by GraphPad Prism 6.0 (GraphPad Software, USA), parametric analysis using Anova test, Tukey post-test and Kruskal-Wallis non-parametric statistical test, followed by t student test were adopted for determination of differences between the experimental groups. The significance index considered in this work was 5%. For both cell viability assessment techniques (Alamar Blue and Live/dead assay) analyzed in this work, POH was able to reduce the cell viability of tongue CEO lines in a dose-dependent and time-dependent manner (p<0 .05). The concentrations of 1.5 mM and 3 mM of POH obtained better or similar results to those found in 40 µM cisplatin, for the two strains, in the evaluation of cell viability (p<0.05). The IC50 values of POH were 1.5 mM for the SCC-25 cell at all time intervals (24 h, 48 h and 72 h), since for the HSC-3 line they were 3 mM for 24 h and 48 h times and 1.5 mM for the 72 h interval. POH was able to inhibit the migration of the two DSC cell lines in a concentration-dependent manner (p≤0.05), compared to the control group. The ability of the POH molecule to bind to proteins responsible for cell cycle activation was evaluated using docking models. Among them, the protein GTPase Kras showed the best binding energy (-86.70 kcal/mol), featuring hydrogen bonds with residues THR58 (A) and ASP57 (A) and steric bonds with residues TRY32 (A) and ALA18 ( THE). The evidence from this study supports the idea that POH has antineoplastic activity on the CEO, suggesting that this molecule may be a strong candidate for the development of drugs aimed at the treatment of this pathology (AU).

A comprehensive profile of TCF1+ progenitor and TCF1- terminally exhausted PD-1+CD8+ T cells in head and neck squamous cell carcinoma: implications for prognosis and immunotherapy / 国际口腔科学杂志·英文版

The heterogeneity of exhausted T cells (Tex) is a critical determinant of immune checkpoint blockade therapy efficacy. However, few studies have explored exhausted T cell subpopulations in human cancers. In the present study, we examined samples from two cohorts of 175 patients with head and neck squamous cell cancer (HNSCC) by multiplex immunohistochemistry (mIHC) to investigate two subsets of Tex, CD8+PD1+TCF1+ progenitor exhausted T cells (TCF1+Texprog) and CD8+PD1+TCF1- terminally exhausted T cells (TCF1-Texterm). Moreover, fresh tumor samples from 34 patients with HNSCC were examined by flow cytometry and immunohistochemistry to further investigate their properties and cytotoxic capabilities and their correlation with regulatory T cells (Tregs) in the tumor immune microenvironment (TIME). mIHC and flow cytometry analysis showed that TCF1-Texterm represented a greater proportion of CD8+PD1+Tex than TCF1+Texprog in most patients. TCF1+Texprog produced abundant TNFα, while TCF1-Texterm expressed higher levels of CD103, TIM-3, CTLA-4, and TIGIT. TCF1-Texterm exhibited a polyfunctional TNFα+GZMB+IFNγ+ phenotype; and were associated with better overall survival and recurrence-free survival. The results also indicated that larger proportions of TCF1-Texterm were accompanied by an increase in the proportion of Tregs. Therefore, it was concluded that TCF1-Texterm was the major CD8+PD1+Tex subset in the HNSCC TIME and that these cells favor patient survival. A high proportion of TCF1-Texterm was associated with greater Treg abundance.

Oral Squamous Cell Carcinoma Associated with Arsenic Exposure: A Case Series from Argentina / Carcinoma Oral de Células Escamosas Asociado con la Exposición al Arsénico: Una Serie de Casos de Argentina

ABSTRACT: Many areas of South America are directly affected by Arsenic (As) contaminated groundwater. A high percentage of the water samples taken in multiple areas of Argentina had As concentrations above the WHO level recommended guidelines. This condition was previously associated with an increased risk of chronic diseases, including different cancers. Long-term As exposure was proposed as a risk factor for Oral Squamous Cell Carcinoma (OSCC). The aim of this study is to present a series cases of Argentine patients diagnosed with OSCC who have consumed water contaminated with As for more than 10 years. Clinical data were obtained from the archive of Clinical Records Histories of the Oral Medicine Department of the Dentistry School, Universidad Nacional de Córdoba and Universidad Católica de Córdoba, Argentina. 15 cases of OSCC were included. The male: female sex ratio was 2:1. The average age was 66 years (31-89 years). Regarding location, the gum or edentulous alveolar ridge was the most prevalent site (6/15; 40 %), followed by the tongue margin. The average years of exposure to arsenical waters were 24 years (13 - 40 years of exposure). The majority of the presented cases were non drinkers non smokers. 60 % of the tumors were diagnosed at advanced stages. the epidemiological studies carried out in As-contaminated areas that address oral cancer should always incorporate the record of variables related to As exposure. Patients who live or lived at As-contaminated areas must be regularly followed up for early diagnosis of potentially malignant or malignant lesions. The high frequency of gum cancer among these cases, should raise awareness of periodontic specialists to perform a careful and thorough periodontal examination.

RESUMEN: Muchas regiones de América del Sur están directamente afectadas por aguas subterráneas contaminadas con arsénico (As). Un alto porcentaje de las muestras de agua tomadas en múltiples áreas de Argentina tenían concentraciones de As superiores al nivel aprobado por la OMS. Esta condición se asociaba previamente con un mayor riesgo de enfermedades crónicas, incluidos diferentes tipos de cáncer. La exposición a largo plazo se propuso como un factor de riesgo para el carcinoma oral de células escamosas (OSCC). El objetivo de este estudio es presentar una serie de casos de pacientes diagnosticados con OSCC que han consumido agua contaminada con As durante más de 10 años. Se obtuvieron datos clínicos del archivo de Historias de registros clínicos del Departamento de Medicina Oral de la Facultad de Odontología, Universidad Nacional de Córdoba y Universidad Católica de Córdoba, Argentina. Se incluyeron 15 casos de OSCC. La relación de género masculino: femenino fue de 2: 1. La edad promedio fue de 66 años (31-89 años). En cuanto a la ubicación, la encía o la cresta alveolar edéntula fue el sitio más frecuente (6/15; 40 %), seguido del borde de la lengua. El promedio de años de exposición a las aguas arsenicales fue de 24 años (13 - 40 años de exposición). La mayoría de los casos presentados fueron de pacientes no bebedores y no fumadores. El 60 % de los tumores fueron diagnosticados en etapas avanzadas. Los estudios epidemiológicos realizados en áreas contaminadas con As que abordan el cáncer oral siempre deben incorporar el registro de variables relacionadas con la exposición a As. Se debe hacer un seguimiento continuo de los pacientes que viven o que vivieron en áreas contaminadas con As para el diagnóstico temprano de lesiones potencialmente malignas. La alta frecuencia de cáncer de encías en estos casos, debe concienciar a los especialistas en periodoncia para que realicen un examen periodontal cuidadoso y completo.

Cociente ganglionar como factor pronóstico de sobrevida. Recurrencia en carcinoma escamoso de cavidad oral / NODAL RATIO AS A PROGNOSTIC FACTOR OF SURVIVAL AND RECURRENCE IN ORAL SQUAMOUS CELL CARCINOMA

Antecedentes: El carcinoma escamoso de cavidad oral constituye una de las patologías de mayor frecuencia en cabeza y cuello. El tratamiento de elección es quirúrgico con altas tasas de control local. Sin embrago, se evidencian recidivas aún en etapas tempranas de la enfermedad, lo que lleva a la necesidad de identificar factores pronósticos confiables para mejorar la estadificación, tratamiento y seguimiento. El Cociente Ganglionar fue ratificado como herramienta pronóstica en otros tumores y validado en cavidad oral en un estudio multicéntrico dirigido por el Memorial Sloan Kettering Center de Nueva York. Objetivo: Validar en nuestro medio al Cociente ganglionar como factor pronóstico de sobrevida y recurrencia en carcinoma de cavidad oral. Lugar de aplicación: Hospital público de atención terciaria de tumores. Diseño: Cohorte Retrospectivo Institucional. Material y Métodos: Se recabó de manera retrospectiva información de historias clínicas, partes quirúrgicos e informes de anatomía patológica de un total de 92 pacientes. Se incluyeron aquellos con carcinoma escamoso de cavidad oral T 1-4 pN0-pN+ (pN1-2). Se calculó la sobrevida utilizando el método Kaplan-Meier y se realizó el análisis multivariado. Resultados: Un Cociente Ganglionar (CG.) mayor a 5% resultó estadísticamente significativo como factor pronóstico de sobrevida [HR 5,22(IC95% 1,86; 14;62) (p 0.002)] y recurrencia [HR 13.33 (IC95% 3.85; 46.16) No se evidenció diferencia pronóstica entre aquellos pacientes con vaciamientos pN0 y pN+ asociado a CG. menor a 5%. Pacientes pN1 y vaciamientos con recuento ganglionar total de 20 (1/20) podrían obtener un similar pronóstico al de pN0. Conclusiones: Fue posible ratificar al CG. como factor pronóstico y se plantea la posible utilidad del mismo en la indicación del tratamiento adyuvante

Background: Oral cavity squamous cell carcinoma is one of the most common pathologies in head and neck surgery. Surgery is the treatment of choice with high rates of locoregional control. However, recurrences are seen even in early stages of the disease. This explains the need of new prognostic factors to provide a better staging, treatment and follow up. Nodal Ratio has been validated as a prognostic tool in other tumors and in oral cavity in a multicentric study by Memorial Sloan Kettering Center of Nueva York. Objective: To validate the Nodal Ratio as a prognostic factor in terms of survival and recurrence in oral cavity carcinoma in our medium. Design: Institutional retrospective cohort. Setting: Tertiary Public Hospital for treatment of tumours. Population and Methods: We retrospectively review 92 patient´s information from clinic histories, surgical protocols and pathologic informs. Inclusion criteria were squamous cell histopathology, T1-4 and pN0-N+ (pN1-2) stage. We calculate survival with Kaplan-Meier method and multivariate analysis was done to determinate the independence value. Results: A Nodal Ratio higher than 5% was statistically significant as a prognostic factor of survival [HR 5,22(IC95% 1,86; 14;62) (p 0.002)] and recurrence [HR 13.33 (IC95% 3.85; 46.16) There was no prognostic difference between patients with pN0 dissections and those with pN+ dissections plus NR less than 5%. pN1 patients with nodal yields of 20 (1/20) or more could have the same forecast as a pN0. Conclusions: We were able to validate NR as a prognostic factor. We postulate the potential use in the indication of adjuvant therapy.

Beta-adrenergic pathway activation in head and neck cancer: a comprehensive analysis of the clinical and genomic features using integrated bioinformatics / Ativação da via beta-adrenérgica no câncer de cabeça e pescoço: uma análise abrangente das características clínicas e gênomicas utilizando a bioinformática

O estresse crônico leva à ativação da via de sinalização beta-adrenérgica. Sua ativação tem sido implicada na progressão de diferentes tipos de câncer, mas seu papel nos carcinomas espinocelulares de cabeça e pescoço (CECPs) permanece indefinido. O objetivo deste estudo foi investigar o papel da ativação da via betaadrenérgica na progressão dos CECPs, avaliar seu impacto na sobrevida dos pacientes e buscar possíveis terapias para pacientes que encontravam-se com a via beta-adrenérgica ativa. Quinhentos e vinte pacientes do The Cancer Genome Atlas com CECPs primários foram divididos em dois grupos: ADRB2baixa / SLC6A2baixa e ADRB2alta / SLC6A2alta. A associação de características clinicopatológicas e genômicas entre os grupos foram analisadas utilizando bioinformática. Os genes diferencialmente expressos (DEGs) foram identificados através da análise da expressão diferencial. A análise de sobrevida também foi realizada com base nas expressões ADRB2 e SLC6A2. Foram identificados medicamentos em potencial para tratamento de CECPs com base nos DEGs. Houve associação entre as expressões ADRB2 e SLC6A2 com idade, raça, localização do tumor, grau histológico, invasão perineural e status do HPV p16. Foram identificados 898 DEGs entre os grupos. Foi demonstrado que a expressão ADRB2alta / SLC6A2alta influenciou a proliferação, adesão e invasão de células CECPs além da angiogênese. Pacientes com carcinomas espinocelular de laringe e faringe apresentando expressão ADRB2alta / SLC6A2alta tiveram menor sobrevida. Por fim, 56 drogas antineoplásicas e imunoterápicas aprovadas pelo Food Drugs Administration foram identificadas como potenciais alvos para o tratamento personalizado. Significância: Estes achados sugerem fortemente um papel proeminente da sinalização beta-adrenérgica no CECPs ao estimular um fenótipo tumoral mais agressivo. Estas alterações tiveram um impacto negativo no prognóstico dos pacientes com CECP em região de faringe e laringe(AU)

Chronic stress leads to the activation of the beta-adrenergic pathway. Its activation has been implicated in the progression of different types of cancer but its role on head and neck squamous cell carcinomas (HNSCCs) remains undefined. The aim of this study was to investigate the influence of the beta-adrenergic pathway activation in the progression of HNSCCs, assess its impact in the survival of the patients, and explore the potential targets. Five hundred and twenty The Cancer Genome Altas patients with primary HNSCCs were divided in two groups: ADRB2low / SLC6A2low and ADRB2high / SLC6A2high. The association of clinicopathological and genomic features between the groups was analyzed using a bioinformatic approach. Differentially expressed genes (DEGs) were identified through differential expression analysis. Survival analysis was also performed based on ADRB2 and SLC6A2 expressions. Potential drugs for treatment of HNSCC were identified based on the DEGs. There was association between ADRB2 and SLC6A2 expressions with age, race, tumor site, histologic grade, perineural invasion, and HPV p16 status. It was identified 898 DEGs between the groups. It was demonstrated that ADRB2high / SLC6A2high expression influenced HNSCC cells proliferation, adhesion, invasion, and angiogenesis. Patients with larynx and pharynx squamous cell carcinomas presenting ADRB2high / SLC6A2high expression showed had lower survival rates. Finally, 56 Food Drugs Administration-approved antineoplastic and immunotherapeutic drugs were identified as potential targets for the personalized treatment. Significance: These findings strongly suggest a prominent role of beta-adrenergic pathway in HNSCC by stimulating a more aggressive tumoral phenotype. These alterations were shown to negatively impact the prognosis of patients with larynx and pharynx squamous cell carcinomas(AU)