Evaluation of tumor load in sentinel lymph node in patients with cutaneous melanoma / Avaliação da carga do tumor em linfonodo sentinela nos doentes portadores de melanoma cutâneo

ABSTRACT Introduction: cutaneous melanoma (MC) is a malignant neoplasm derived from melanocytic cells with an aggressive behavior. It is usually associated with the multifactorial interaction of genetic susceptibility and environmental exposure, usually ultraviolet radiation. Despite advances in treatment, the disease remains relentless with poor prognosis. Sentinel lymph node (SLN) biopsy is a technique used to screen patients in need of lymph node dissection. Objectives: to correlate the tumor burden in the SLN with the mortality of patients undergoing SLN biopsy. Methodology: the medical records and histological slides of patients with MC who underwent SLN biopsy treated at HC-Unicamp from 2001 to 2021 were retrospectively analyzed. The positive SLN were measured according to the size of the tumor infiltration area, for analysis of the depth of invasion (DI), closest proximity to the capsule (CPC) and tumor burden (TB). For statistical analysis, associations between variables were analyzed using Fishers exact test, with post Bonferroni test and Wilcoxon test. Results: 105 records of patients who underwent SLN biopsy of MC were identified. Of these, nine (8.6%) had positive SLN and 81 (77.1%) had negative SLN. The performed lymphadenectomies resulted in 55.6% (n=5) affected, 22.2% (n=2) without disease and 22.2% (n=2) were not performed. Mean CPC, TB, and DI were 0.14mm, 32.10mm and 2.33mm, respectively. Patients with T2 and T3 tumors were more likely to show the SLN affected (p=0.022). No patient with positive SLN died during follow-up. Conclusion: patients who presented T3 staging are the ones who most presented positive SLN.

RESUMO Introdução: o melanoma cutâneo (MC) é uma neoplasia maligna de comportamento agressivo, derivada das células melanocíticas, geralmente causado pela associação de interação da suscetibilidade genética e a exposição ambiental. A biópsia do linfonodo sentinela (LNS) é um procedimento utilizado para rastreamento de doentes com necessidade ou não de linfadenectomia, diminuindo a exposição do paciente a cirurgias maiores. Objetivos: correlacionar a carga do tumor no LNS com a mortalidade de pacientes com MC. Métodos: foram examinados retrospectivamente prontuários e lâminas histológicas de doentes com MC submetidos a biópsia de LNS atendidos no HC-Unicamp entre o período compreendido de 2001 a 2021. Os LNS positivos foram mensurados quanto ao tamanho da área de infiltração do tumor, para análise da profundidade de invasão (PI), menor proximidade com a cápsula (MPC) e carga do tumor (CT). As associações entre as variáveis foram analisadas pelo teste Exato de Fisher, com pós teste de Bonferroni e Wilcoxon. Resultados: foram identificados 105 pacientes com biópsia de LNS, sendo nove (8,6%) casos com LNS positivo. A média de MDC, CT e PI foram 0,14 mm, 32,10 mm, e 2,33 mm, respectivamente. Doentes com tumores T2 e T3 apresentaram maior chance de acometimento do LNS (p=0,022). Nenhum paciente com LNS positivo foi a óbito durante o acompanhamento. Conclusão: doentes que apresentaram tumores maiores (T2 e T3) apresentaram maior chance de linfonodos sentinelas positivos. Além do tamanho do tumor, a presença de maior carga do tumor demonstrou que os mesmos podem ser beneficiados pela biópsia de LNS.

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

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.

D2-40/podoplanin expression in chronic villitis of the placenta / Expressão de podoplanina/D2-40 em vilosite crônica placentária

ABSTRACT D2-40/podoplanin (D2-40/PDPN) is a multifunctional protein that can be expressed in lymphatic endothelium and immune cells. D2-40/ PDPN expression in chronic villitis (CV) has not been studied. In 22 cases of CV, we analyzed both D2-40/PDPN expression as well as its coexpression with immune cells markers, and the relationship with stromal cells. In the non-inflamed villi, the D2-40/PDPN positive plexiform pattern has a lymphatic-like conductive network. In the inflamed villi, the D2-40/PDPN expression, predominantly restricted to stromal cells forming a cellular network, is likely related to a phase of the inflammatory response, such as reorganization of the damaged tissue.

RESUMO Podoplanina/D2-40 (PDPN/D2-40) é uma proteína multifuncional que pode ser expressa no endotélio linfático e nas células imunes. Na vilosite crônica (VC), a expressão de PDPN/D2-40 ainda não foi estudada. Em 22 casos de VC, analisamos tanto a expressão de PDPN/D2-40 como sua coexpressão com marcadores de células imunes, além da relação com células estromais. Nas vilosidades não inflamadas, o padrão plexiforme PDPN/D2-40 positivo tem aspecto de rede condutora linfática. Nas vilosidades inflamadas, a expressão de PDPN/D2-40, com predominância restrita às células estromais, formando rede densa está, possivelmente, relacionada com uma fase da resposta inflamatória, como a reorganização do tecido danificado.

Carcinoma ex-pleomorphic adenoma derived from recurrent pleomorphic adenoma shows important difference by array CGH compared to recurrent pleomorphic adenoma without malignant transformation / Carcinoma ex-adenoma pleomórfico derivado de adenoma pleomórfico recorrente mostra diferença importante por array CGH em comparação com adenoma pleomórfico recorrente sem transformação maligna

Abstract Introduction: A key step of cancer development is the progressive accumulation of genomic changes resulting in disruption of several biological mechanisms. Carcinoma ex-pleomorphic adenoma (CXPA) is an aggressive neoplasm that arises from a pleomorphic adenoma. CXPA derived from a recurrent PA (RPA) has been rarely reported, and the genomic changes associated with these tumors have not yet been studied. Objective: We analyzed CXPA from RPAs and RPAs without malignant transformation using array-comparative genomic hybridization (array-CGH) to identify somatic copy number alterations and affected genes. Methods: DNA samples extracted from FFPE tumors were submitted to array-CGH investigation, and data was analyzed by Nexus Copy Number Discovery Edition v.7. Results: No somatic copy number alterations were found in RPAs without malignant transformation. As for CXPA from RPA, although genomic profiles were unique for each case, we detected some chromosomal regions that appear to be preferentially affected by copy number alterations. The first case of CXPA-RPA (frankly invasive myoepithelial carcinoma) showed copy number alterations affecting 1p36.33p13, 5p and chromosomes 3 and 8. The second case of CXPA-RPA (frankly invasive epithelial-myoepithelial carcinoma) showed several alterations at chromosomes 3, 8, and 16, with two amplifications at 8p12p11.21 and 12q14.3q21.2. The third case of CXPA-RPA (minimally invasive epithelial-myoepithelial carcinoma) exhibited amplifications at 12q13.3q14.1, 12q14.3, and 12q15. Conclusion: The occurrence of gains at chromosomes 3 and 8 and genomic amplifications at 8p and 12q, mainly those encompassing the HMGA2, MDM2, WIF1, WHSC1L1, LIRG3, CDK4 in CXAP from RPA can be a significant promotional factor in malignant transformation.

Resumo Introdução: Uma etapa fundamental do desenvolvimento do câncer é o acúmulo progressivo de alterações genômicas, resultando na ruptura de vários mecanismos biológicos. Carcinoma ex-adenoma pleomórfico (CXAP) é uma neoplasia agressiva que surge a partir de um adenoma pleomórfico. O CXAP derivado de um AP recorrente (APR) foi raramente relatado e, até o momento, as alterações genômicas associadas a esses tumores não foram estudados. Objetivo: Avaliar as diferenças entre os CXAPs decorrentes de APRs e os APRs sem transformações malignas usando hibridização genômica comparativa em microarrays (array Comparative Genomic Hibridization - aCGH) a fim de identificar alterações no número de cópias somáticas e os genes afetados. Método: Amostras de DNA extraídas de tumores provenientes de tecido emblocado em parafina foram submetidos à investigação com a técnica aCGH, e os dados foram analisados com o Nexus Copy Number Discovery Edition v.7. Resultados: Não observamos alterações no numero de cópias somáticas nos APRs sem transformação maligna. Quanto ao CXAP de APR, embora os perfis genômicos sejam exclusivos para cada caso, detectamos algumas regiões cromossômicas que pareciam ser preferencialmente afetadas por alterações no número de cópias. O primeiro caso de CXAP-APR (carcinoma mioepitelial francamente invasivo) apresentou alterações no numero de cópias afetando 1p36.33p13, 5p e cromossomos 3 e 8. O segundo caso de CXAP-APR (carcinoma epitelialmioepitelial francamente invasivo) apresentou várias alterações nos cromossomos 3, 8 e 16, com duas amplificações em 8p12p11.21 e 12q14.3q21.2. O terceiro caso de CXAP-APR (carcinoma epitelial-mioepitelial minimamente invasivo) apresentou amplificações em 12q13.3q14.1, 12q14.3, e 12q15. Conclusão: A ocorrência de ganhos de cromossomos 3 e 8, e as amplificações genômicas em 8p e 12q, principalmente aquelas que englobam os HMGA2, MDM2, WIF1, WHSC1L1, RG3, CDK4 no CXAP decorrente de APR podem ser fatores promocionais significativos para a transformação maligna.

Cellular Proliferation Index between Carcinoma Ex-Pleomorphic Adenoma and Pleomorphic Adenoma

Carcinoma ex pleomorphic adenoma (CXPA) has been considered an interesting model of carcinogenesis, presenting various histological subtypes and invasiveness phase. The objective was to determine the proliferative index of CXPA and comparing to pleomorphic adenoma (PA). Thirty six cases of CXPA (36 PA) and 22 areas of PA in CXPA (residual PA) were studied by Ki-67 expression. All CXPA cases were classified according to invasiveness phase (intracapsular, minimally and frankly invasive) and histopathological subtypes. Data was statistically analyzed by Wilcoxon, Mann-Whitney and Kruskal-Wallis tests. CXPA included 5 intracapsular, 9 minimally invasive and 22 frankly invasive cases. Fifteen cases corresponded to salivary duct carcinoma, 7 to adenocarcinoma NOS, 7 myoepithelial, 5 epithelial-myoepithelial, one case of squamous cell and one case of sarcomatoid carcinoma. The Ki-67 index of PA and residual PA were significantly lower than CXPA. Intracapsular and minimally invasive showed smaller proliferative index than frankly invasive. Considering the subtypes of CXPA, there was not a statistic difference among them. Ki-67 is a useful marker in the differential diagnosis of PA and CXPA, even when in the early invasive phase.

Carcinoma ex adenoma pleomorfo (CXAP) tem sido considerado um interessante modelo de carcinogênese, apresentando vários subtipos histológicos e fases de invasividade. Determinar o índice proliferativo de CXAP e compará-lo ao adenoma pleomorfo (AP). e seis casos de CXAP, 36 AP, e 22 áreas de AP em CXAP (AP residual) foram estudadas através da expressão de Ki-67. Todos os casos de CXAP foram classificados de acordo com a fase de invasividade (intracapsular, minimamente invasivo e francamente invasivo) e de acordo com os diversos subtipos histopatológicos. Os dados foram estatisticamente analisados através dos testes Wilcoxon, Mann-Whitney e Kruskal-Wallis. O grupo de CXAP era formado por 5 intracapsulares, 9 minimamente invasivos e 22 francamente invasivos. Quinze casos corresponderam a carcinoma de ducto salivar, 7 a adenocarcinoma nos, 7 a carcinoma mioepitelial, 5 a carcinoma epitelial-mioepitelial, 1 a carcinoma epidermoide e 1 a carcinoma sarcomatóide. Os índices de Ki-67 de AP e AP residual foram significativamente menores que o encontrado em CXAP. Os casos intracapsulares e minimamente invasivos mostraram índices proliferativos menores que os francamente invasivos. Considerando os subtipos histológicos de CXAP, não houve diferença estatística entre eles. Ki-67 é um marcador útil no diagnóstico diferencial de AP e CXAP, mesmo quando o carcinoma está em fase precoce de invasividade.