P53 And Expression Of Immunological Markers May Identify Early Stage Thyroid Tumors.

Besides its major role in cell proliferation, DNA repair, and apoptosis, functional p53 protein is involved in the induction of antitumor cytotoxic-T-cell activity against carcinoma cells. We aimed to investigate p53 and immune cell markers utility as diagnostic and prognostic markers of differentiated thyroid cancer (DTC). ACIS-III system was used to evaluate p53 and immune cell markers including tumor-associated macrophages (TAM); CD68 and tumor-infiltrating lymphocytes (TIL) subsets such as CD3, CD4, CD8, and CD20 in 206 thyroid carcinomas, 105 benign nodules, and 18 normal tissues. Also, TP53 was sequenced in 78 out of 164 patients with papillary thyroid carcinoma. P53 expression was observed more frequently in malignant than in benign lesions (P < 0.0001) and helped discriminate follicular patterned lesions. In addition, p53 was more frequent in smaller (P = 0.0015), unique tumors (P = 0.0286), with thyroiditis (P = 0.0486) and without metastasis at diagnosis (P = 0.0201). TAM was more frequent in P53 negative tumors (P = 0.002). Infiltration of CD8+ TIL was found in 61.7% of P53 positive and 25.6% of P53 negative DTC (P < 0.001). We suggest that p53 and CD8+ TIL immune profile analysis might be useful in DTC.201384658

Molecular profile of adiponectin, ghrelin, leptin and resistin : esteblishing the links between obesity and thyroid cancer

Orientador: Laura Sterian WardTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: A incidência da obesidade, bem como do câncer diferenciado de tireoide (CDT), tem aumentado exponencialmente em todo o mundo e existem sólidos indícios de que essas duas condições se associam. Diversos estudos demonstram que adipocinas como adiponectina, leptina e resistina e também a grelina podem ser o elo entre essas doenças. O objetivo deste trabalho foi de verificar os perfis antropométrico e molecular das adipocinas: adiponectina, leptina, resistina e grelina de pacientes com diversos tipos de nódulos tireoidianos e compará-los a indivíduos saudáveis, buscando encontrar o elo entre a obesidade e o CDT, além de identificar possíveis marcadores de diagnóstico, evolução e prognóstico para o CDT. Estudamos pacientes com CDT, pacientes com nódulos tireoidianos benignos e indivíduos controle saudáveis. Todos foram entrevistados quanto a seus hábitos alimentares e rotina, além de passarem por avaliação antropométrica completa. Para investigar o perfil molecular da adiponectina, leptina, resistina e grelina, investigamos as concentrações séricas dessas citocinas através da técnica de ELISA, genotipamos os pacientes para 21 SNPs nos genes que codificam as adipocinas e tambem seus receptores através da técnica Taqman SNP Genotyping, analisamos ainda, espécimes cirúrgicos dos pacientes quanto a produção de RNAm dessas adipocinas. Demonstramos que o excesso de peso aumenta o risco de desenvolvimento de CDT em quase 4 vezes e esse risco pode aumentar ainda mais com a ingesta excessiva de calorias, associada ao consumo excessivo de carboidratos e proteínas. As concentrações séricas das adipocinas podem ser excelentes marcadores de diagnostico entre nódulos tireoidianos malignos e benignos, sendo que a acurácia do teste utilizando a leptina e resistina chegou a 100%. A herança genetica dos polimorfismos rs7799039 em LEP, rs1137101 em LEPR e rs2232165 em GHSR aumentou o risco de desenvolvimento de CDT em 4, 11 e 22 duas vezes, respectivamente. Além disso, a herança de SNPs em LEP (rs7799039, rs2167270), ADIPOQ - rs12629945, ADIPOR1 ¿ rs2232853 e rs1342387, ADIPOR2 - rs1058322 e GHSR - rs2232165 alterou as concentrações séricas de leptina, adiponectina e grelina. Demonstramos, ainda, que embora a célula tireoidiana não seja a principal produtora de adipocinas, a adiponectina e seus receptores, o receptor da leptina, e a grelina e seu receptor são produzidos por células tumorais tireoidianas, uma vez que encontramos RNAm para todas essas citocinas nos tecidos analisados, sugerindo que essas adipocinas desempenham um papel importante nesses tecidos. Concluímos que indivíduos portadores de nódulos tireoidianos, especialmente os indivíduos portadores de CDT possuem um perfil antropométrico e molecular (em relação às adipocinas) diferenciado, sendo que aqueles que estão mais fora do peso considerado adequado estão mais suscetíveis ao desenvolvimento do CDT, assim como aqueles que apresentam alterações genéticas e/ou de expressão da adiponectina, leptina, resistina e grelina. Sugerimos que é possível traçar um perfil do individuo portador de CDT em relação a sua constituição física, seus hábitos alimentares e também o seu perfil molecular a adiponectina, leptina, resistina e grelina, que foram identificadas como boas candidatas a marcadores moleculares para o CDT, além de serem potenciais links entre a obesidade, inflamação e CDTAbstract: The incidence of obesity and of differentiated thyroid cancer (DTC) has increased exponentially over the world and there are strong indications that these two conditions are combined. Several studies have shown that adipokines such as adiponectin, resistin, leptin and ghrelin may be the link between these diseases. The objective of this investigation was to study the molecular profile of adipokines: adiponectin, leptin, resistin and ghrelin and its relationship with the anthropometric features of patients with various types of thyroid nodules in order to investigate the link between obesity and neoplasms and identify possible markers of diagnosis, evolution and prognosis for DTC. A group of patients with DTC, patients with benign thyroid nodules and healthy control subjects were interviewed about their eating habits and routine, and went through a complete anthropometric assessment. The serum concentrations of adiponectin, leptin, resistin, and ghrelin were measured by ELISA. Patients were genotyped for 21 SNPs in genes encoding adipokines and their receptors using Taqman SNP Genotyping technique. In addition, surgical specimens were investigated for mRNA of these adipokines. We demonstrated that being overweight increases the risk of developing CDT almost 4 times and this risk may increase when there is excess intake of calories associated with excessive consumption of carbohydrates and proteins. Serum concentrations of adipokines were excellent markers of nodules malignancy diagnosis, with a 100% accuracy using leptin and resistin. The genetic inheritance of polymorphisms rs7799039 in LEP rs1137101 and rs2232165 in LEPR in GHSR increased the risk of developing CDT 4, 11 and 22 times, respectively. In addition, SNPs heritage in LEP (rs7799039, rs2167270), ADIPOQ - rs12629945, ADIPOR1 - rs2232853 and rs1342387, ADIPOR2 - rs1058322 and GHSR - rs2232165 altered sérum concentrations of leptin, adiponectin and ghrelin. Also, we showed that although the thyroid cell is not the main producer of adipokines, adiponectin, leptin, and ghrelin resitina are produced by thyroid câncer cells, findng mRNA for all cytokines in the tissues analyzed, suggesting that they play an important role in tumorigenesis or thyroid tumor proliferation. We conclude that individuals with thyroid nodules, especially individuals with DTC have diferente anthropometric and molecular adipokine profiles that increases the susceptibility to DTC as well as genetic changes and / or expression of adiponectin, leptin, resistin and ghrelin. We suggest that it is possible to draw a CDT carrier Individual profile for body constitution, eating habits and molecular profile of adiponectin, leptin, resistin and ghrelin may identify individuals at risk for DTC, and are a potential link between obesity, inflammation, and DTCDoutoradoClinica MedicaDoutora em Clínica Médic

Malignancy markers for thyroid nodules

Orientadores: Laura Sterian Ward, André Lopes CarvalhoDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: Nódulos de tiróide são palpados em cerca de 10% da população e diagnosticados em mais de 50% da população usando-se a ultrassonografia. No entanto, apenas 0,01% são câncer. Por outro lado, o Câncer Diferenciado da Tiróide (CDT) é a neoplasia endócrina mais comum e suas taxas de incidência vem crescendo pelo mundo. Urge, portanto, estabelecer marcadores capazes de identificar malignidade em nódulos na população. Objetivamos, neste trabalho, investigar a utilidade clínica de BRAF, TPO, GAL-3, VEGF e VEGF-C como marcadores para o CDT. Foram utilizados 165 pacientes com CDT; 31 pacientes com nódulos Benignos e 216 Controles. Utilizamos ensaios de PCR-RFLP para a pesquisa da mutação de BRAF, RT-PCRq para dosar TPO e ELISA para dosar VEGF, VEGF-C e Galectina-3 circulantes. A identificação da mutação de BRAF em células tiroidianas circulantes não foi possível com a metodologia por nós empregada. Não encontramos expressão de TPO em sangue periférico de pacientes com CDT. O nível sérico de Gal-3 diferiu entre os grupos CDT, Benigno e Controles, mas não diferenciou nódulos benignos de malignos (p=0, 884). O nível sérico de VEGF pôde diferenciar os pacientes com CDT de pacientes com nódulos Benignos (p=0, 041), assim como diferenciou CDT dos controles (p=0.015), embora não tenha grande poder como marcador único. Os níveis séricos de VEGF-C distinguem pacientes com nódulos Benignos, Malignos e Controles (p=0, 025). Dentre estes, distinguem pacientes com CDT dos Controles (p=0, 046), porém não dos nódulos Benignos (p=0.242). Os níveis séricos de Gal-3, VEGF e VEGF-C parecem discriminar alguns grupos em relação a variáveis clínico-patologicas, porem não há indicação de que possam ser usados como marcadores para o CDT, uma vez que seus poderes preditivos positivo e negativo não são bons. Concluímos que, embora possam auxiliar no diagnóstico, nenhum dos marcadores estudados é promissor do ponto de vista clínicoAbstract: Thyroid nodules may be diagnosed in about 10% of the population and half of the population if they're examined through ultrasonography. However, only 0.1% of those nodules are cancer. On the other hand, Differentiated Thyroid Cancer (DTC) is the most common endocrine malignancy and its incidence rate is increasing worldwide. This fact creates an urgent need for new markers that might be used in a large scale to diagnose thyroid nodules malignancy. We aimed to investigate the clinical utility of BRAF, TPO, Gal-3, VEGF and VEGF-C as markers for the CDT. We studied 165 patients with DTC, 31 patients with benign tumors and 216 controls. We used PCR-RFLP to investigate the presence of BRAF mutation, RT-PCRq to quantify TPO mRNA and ELISA to quantify VEGF, VEGF-C and Gal-3 in circulation. The identification of BRAF mutation in thyroid cells was not possible with the methodology employed. We didn't find any expression of TPO in peripheral blood of patients with DTC. The serum levels of Gal-3 differed among groups DTC, Benign and Controls, but did not differentiate benign and malignant tumors (p = 0.884). The serum levels of VEGF could differentiate patients with DTC from patients with benign nodules (p = 0.041) and DTC differed from controls (p = 0.015), although this test didn't have a great predictive power as a single marker. Serum levels of VEGF-C distinguished patients with benign nodules, malignant tumors and controls (p = 0.025). It distinguished patients with DTC from Controls (p = 0.046) but not from the benign tumors (p = 0242). Serum levels of Gal-3, VEGF and VEGF-C seem to discriminate some groups in what concerns clinical and pathological variables, but there is no indication that they can be used as markers for the CDT, as its positive and negative predictive powers are not high. We conclude that, although they might support the diagnosis, none of the markers studied is promising in a clinical perspectiveMestradoCiencias BasicasMestre em Clinica Medic

Obesity And Thyroid Cancer.

Many studies have provided observational data on the association of obesity and thyroid cancers, but only few of them propose mechanisms that would permit a better understanding of the causal molecular mechanisms of this association. Considering that there is an increasing incidence of both obesity and thyroid cancers, we need to summarize and link recent studies in order to characterize and understand the contribution of obesity-related factors that might affect thyroid cancer development and progression. Adipose tissue is involved in many vital processes, including insulin sensitivity, angiogenesis, regulation of energy balance, activation of the complement system, and responses such as inflammation. Although these processes have their own molecular pathways, they involve the same molecules through which obesity and adipose tissue might exert their roles in carcinogenesis, not only affecting MAPK and PI3K or even insulin pathways, but also recruiting local inflammatory responses that could result in disease formation and progression. This review describes five important issues that might explain the link between excessive weight and thyroid cancer: thyroid hormones, insulin resistance, adipokines, inflammation, and sexual hormones.21T255-7

Cd8+ Tumour-infiltrating Lymphocytes And Cox2 Expression May Predict Relapse In Differentiated Thyroid Cancer.

There is an increasing rate of papillary thyroid carcinomas that may never progress to cause symptoms or death. Predicting outcome and determining tumour aggressiveness could help diminish the number of patients submitted to aggressive treatments. We aimed to evaluate whether markers of the immune system response and of tumour-associated inflammation could predict outcome of differentiated thyroid cancer (DTC) patients. Retrospective cohort study. We studied 399 consecutive patients, including 325 papillary and 74 follicular thyroid carcinomas. Immune cell markers were evaluated using immunohistochemistry, including tumour-associated macrophages (CD68) and subsets of tumour-infiltrating lymphocytes (TIL), such as CD3, CD4, CD8, CD16, CD20, CD45RO, GRANZYME B, CD69 and CD25. We also investigated the expression of cyclooxygenase 2 (COX2) in tumour cells and the presence of concurrent lymphocytic infiltration characterizing chronic thyroiditis. Concurrent lymphocytic infiltration characterizing chronic thyroiditis was observed in 29% of the cases. Among all the immunological parameters evaluated, only the enrichment of CD8+ lymphocytes (P = 0·001) and expression of COX2 (P =0·01) were associated with recurrence. A multivariate model analysis identified CD8+ TIL/COX2 as independent risk factor for recurrence. A multivariate analysis using Cox's proportional-hazards model adjusted for the presence of concurrent chronic thyroiditis demonstrated that the presence of concurrent chronic thyroiditis had no effect on prognostic prediction mediated by CD8+ TIL and COX2. In conclusion, we suggest the use of a relatively simple pathology tool to help select cases that may benefit of a more aggressive approach sparing the majority of patients from unnecessary procedures

CD8+ tumour‐infiltrating lymphocytes and COX2 expression may predict relapse in differentiated thyroid cancer

There is an increasing rate of papillary thyroid carcinomas that may never progress to cause symptoms or death. Predicting outcome and determining tumour aggressiveness could help diminish the number of patients submitted to aggressive treatments. We aimed to evaluate whether markers of the immune system response and of tumour‐associated inflammation could predict outcome of differentiated thyroid cancer (DTC) patients. Retrospective cohort study. We studied 399 consecutive patients, including 325 papillary and 74 follicular thyroid carcinomas. Immune cell markers were evaluated using immunohistochemistry, including tumour‐associated macrophages (CD68) and subsets of tumour‐infiltrating lymphocytes (TIL), such as CD3, CD4, CD8, CD16, CD20, CD45RO, GRANZYME B, CD69 and CD25. We also investigated the expression of cyclooxygenase 2 (COX2) in tumour cells and the presence of concurrent lymphocytic infiltration characterizing chronic thyroiditis. Concurrent lymphocytic infiltration characterizing chronic thyroiditis was observed in 29% of the cases. Among all the immunological parameters evaluated, only the enrichment of CD8+ lymphocytes (P = 0·001) and expression of COX2 (P =0·01) were associated with recurrence. A multivariate model analysis identified CD8+ TIL/COX2 as independent risk factor for recurrence. A multivariate analysis using Cox's proportional‐hazards model adjusted for the presence of concurrent chronic thyroiditis demonstrated that the presence of concurrent chronic thyroiditis had no effect on prognostic prediction mediated by CD8+ TIL and COX2. In conclusion, we suggest the use of a relatively simple pathology tool to help select cases that may benefit of a more aggressive approach sparing the majority of patients from unnecessary procedures.832246253FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2010/00394-0; 2011/19612-