O 153Sm-EDTMP é um radiofármaco utilizado em medicina nuclear com resultados promissores no alívio da dor metastática. No entanto, pouco se sabe sobre os efeitos do 153Sm-EDTMP em nível celular. O presente trabalho foi conduzido com o intuito de avaliar os efeitos citogenéticos do 153Sm-EDTMP em linfócitos periféricos de pacientes com metástases ósseas (com e sem radio e/ou quimioterapias anteriores) pela da técnica de detecção de aberrações cromossômicas, tanto in vivo como in vitro. Para tanto, as amostras sangüíneas foram coletadas antes e 1 hora após a administração endovenosa do 153Sm- EDTMP (atividade média de 42,53 + 5,31 MBq/kg de peso corpóreo), levando-se em consideração o rápido clearance sangüíneo. Os principais tipos de aberrações cromossômicas estruturais encontrados foram os gaps e quebras, fragmentos acêntricos, anéis cêntricos, double minutes e dicêntricos. A análise estatística mostrou que o único grupo de pacientes que apresentou uma diferença significativa na freqüência de aberrações cromossômicas 1 hora após o tratamento foi o que recebeu prévio tratamento radio e quimioterápico antes da terapia com 153Sm-EDTMP. Quanto a averiguação do número modal de cromossomos e da cinética do ciclo celular, a análise estatística mostrou que não houve diferença significativa entre os grupos analisados, sugerindo que o tratamento com 153Sm-EDTMP não influenciou nesses parâmetros. A molécula carreadora, EDTMP, não teve qualquer influência na indução de aberrações cromossômicas. Em relação aos ensaios in vitro, os dados obtidos de linfócitos periféricos submetidos às diferentes concentrações radioativas de 153Sm-EDTMP (0,046 – 1,110 MBq/mL) de doadores sadios e de pacientes sem prévio tratamento se ajustaram melhor ao modelo de regressão linear (Y=A+BX). O dano cromossômico induzido pelo 153Sm-EDTMP observado in vitro foi cerca de 2 vezes maior do que o encontrado in vivo para o grupo de pacientes sem prévio tratamento. Os dados obtidos mostraram que a terapia com 153Sm-EDTMP induziu uma pequena quantidade de danos citogenéticos em linfócitos periféricos de pacientes 1 hora após sua administração, embora, teoricamente, um efeito estocástico a longo prazo não possa ser descartado. / The 153Sm-EDTMP is a radiopharmaceutical used in nuclear medicine with promising results for the relief of metastatic pain. Therefore, there are few knowledge about the effects of 153Sm-EDTMP at cellular level. The present study was conducted with the aim of evaluating the cytogenetic effects of 153Sm-EDTMP in peripheral lymphocytes from patients with bone metastasis (with and without previous radio and/or chemotherapy) by the chromosome aberration technique, either in vivo or in vitro. For that, the blood samples were collected before and one hour after the endovenous administrations of 153Sm-EDTMP (mean activity of 42.53 + 5.31 MBq/kg body weight), taking into account the rapid blood clearance. The principal types of structural chromosome aberrations found gaps and breaks, acentric fragments centric rings, double minutes and dicentrics. The statistical analysis showed that the group submitted to previous radio and chemotherapy before153Sm-EDTMP administration showed significant difference in chromosome aberrations frequency one hour after the treatment. The analysis of the chromosome modal number and the kinetics of cellular cycle showed no statistical difference among the groups, suggesting that the treatment with 153Sm-EDTMP, did not influence these parameters. The carrier molecule, EDTMP, did not influence the induction of chromosome aberration. In relation to the in vitro assays, the obtained data of peripheral lymphocytes of healthy donors and patients with no previous treatment exposed to different radioactive concentration of 153Sm-EDTMP (0.046 – 1.110 MBq/mL) were better adjusted by linear regression model (Y=A+BX). The chromosome damage induced by 153Sm-EDTMP observed in vitro was about 2 fold higher than that found in vivo for the group of patients with no previous treatment. The obtained data showed that the therapy with 153Sm- EDTMP induced a few quantity of cytogenetic damages in peripheral lymphocytes one hour after its administration in patients, although, theoretically, a long term stochastic effect cannot be disregarded.

Introdução: O câncer de pulmão é um dos tipos mais comuns de câncer, sendo ele responsável pelo maior número de mortes por neoplasias no mundo. Pacientes com câncer de pulmão apresentam alta prevalência de desnutrição, que está relacionada com piora do prognóstico. A desnutrição ocorre em 60% dos pacientes no momento do diagnóstico, levando a um mau prognóstico, independente do estágio do tumor. Avaliação nutricional de pacientes com câncer deve ser realizada à luz de qualquer proposta terapêutica, ainda que esta seja paliativa. Neste sentido, o método validado de Avaliação Subjetiva Global Produzida pelo Próprio Paciente (ASG-PPP) e adaptado por Ottery et al para pacientes oncológicos a partir da Avaliação Subjetiva Global, ajuda a identificar precocemente os pacientes que estão em risco nutricional. A nutrição é fundamental nestes pacientes em doença avançada, pois fornece a quantidade adequada de nutrientes, tem um importante papel psicológico, social, espiritual e cultural, além de ajudar a manter um sentido de autonomia e bem-estar. Objetivo: Relacionar a Capacidade Funcional através do índice de Karnofsky e o estado nutricional, sinais e sintomas avaliados pelo método subjetivo (ASG-PPP), com a sobrevida de pacientes com neoplasia pulmonar mestastática. Pacientes e métodos: Trata-se de um estudo transversal, constituído por 51 pacientes, ambos os sexos, portadores de neoplasia pulmonar com doença metastática em estadiamento IIIb e IV, atendidos em nível ambulatorial e hospitalar no Hospital Santa Rita, Complexo Hospitalar Santa Casa de Porto Alegre. A sobrevida dos pacientes foi verificada ao final do estudo, através de revisão de prontuários e contato telefônico com o responsável pelo paciente. Resultados: Os pacientes apresentaram-se com idade 61,84±10,83 anos (40 a 82 anos); 34 (66,7%) eram gênero masculino e 17 (33,3%) do feminino. A mediana do tempo de sobrevida foi de 186 dias (IC 95%: 158,8; 213,2). Os pacientes internados tiveram sobrevida significativamente inferior (mediana=145 dias (IC 95%=135 - 155) aos que foram atendidos ambulatorialmente (mediana=221 dias (IC 95%=199 - 243). Os pacientes em estágio IV tiveram sobrevida significativamente inferior (mediana=176 dias; IC 95%=127 - 225) àqueles em estágio IIIb (mediana=221; IC 95%=147 - 295). Os pacientes com metástase hepática tiveram sobrevida significativamente inferior (mediana=115 dias; IC 95%=74 - 155) aos sem metástase hepática (mediana=199; IC 95%=174 - 224). Os pacientes com metástase pleural apresentaram sobrevida significativamente superior (mediana=386 dias; IC 95%=129 - 643) aos sem metástase pleural (mediana=174; IC 95%=133 - 214). Não houve diferença estatisticamente significativa entre as curvas de sobrevida dos diferentes estágios nutricionais (teste de log rank; p=0,651). Os acamados tiveram sobrevida significativamente inferior (mediana=115 dias; IC 95%=87 - 143) que os com capacidade física normal (mediana=255 dias; IC 95%=202 - 308). Quanto maior a depleção corporal total, pior a sobrevida do paciente (p=0,014). O aumento de 1 ponto neste item da ASG-PPP (RDI=1,70; IC 95%=1,12 – 2,59) pode aumentar o risco de óbito, em média 70%. O índice de Karnofsky também se associou significativamente com a sobrevida (p=0,016), sendo que para um aumento de 10% na escala de nível de desempenho, o risco de óbito diminuiu em 23% (RDI=0,77; IC 95%=0,76 – 0,79). Os sintomas mais prevalentes que afetaram a alimentação entre os pacientes foram a falta de apetite (66,7%), depressão (51%) e dor (39,2%) e os menos relatados foram dor na boca e vômitos (5,9%). Os fatores que permaneceram associados estatisticamente com o óbito após o ajuste pela análise multivariada de Regressão de Cox foram metástase hepática e internação hospitalar como fatores de risco para óbito e metástase pleural e melhor índice de Karnofsky como fatores protetores. Conclusão: A sobrevida dos pacientes eutróficos foi levemente superior a dos pacientes em risco nutricional e a dos severamente desnutridos. Quanto maior a depleção corporal total e pior o índice de Karnofsky maior o risco de óbito. Não foi encontrada uma associação estatisticamente significativa entre os valores de Hemograma, Albumina Sérica e Contagem Total de Linfócitos (CTL) com o estado nutricional da população em estudo e a sobrevida. / Introduction: Lung cancer is one of the most common cancers in the world, and it is today the most lethal of the neoplastic diseases. Patients with lung cancer show high prevalence of malnutrition, the latter being related to worsening of the prognosis. Malnutrition occurs in 60% of the patients at the moment of diagnosis, leading to a bad prognosis, regardless of the tumor stage. Nutritional evaluation of patients with cancer should be conducted in the light of a therapeutic proposal, even if it’s palliative. In this way, the validated method Scored Patient-Generated Subjective Global Assessment (PG-SGA), adapted by Ottery et al for oncologic patients from the questionnaire of Subjective Global Assessment, helps to precociously identify patients that are in nutritional risk. Nutrition is fundamental in these patients with advanced disease for it supplies the adequate amount of nutrients, performs an important psychological, spiritual and cultural role, besides helping to build a sense of autonomy and well-being. Objective: to relate Functional Capacity through the Karnofsky Index and the nutritional state, signs and symptoms evaluated by the subjective method (PG-SGA), to the survival of patients with metastatic pulmonary neoplasia. Patients and methods: It’s a transversal study, constituted by 51 patients, both genders, carriers of pulmonary neoplasia with metastatic disease on stages IIIb and IV, both in and outpatients at Santa Rita Hospital, Complexo Hospitalar Santa Casa de Porto Alegre. The survival of the patients in the study was assessed at the end of the study, through review of the chart and phone contact with the patient’s keeper. Results: The average age is 61.84 (± 10.83 varying between 40 and 82), being 34 (66.7%) male and 17 (33.3%) female. The median of survival is 186 days (CI 95%; 158.8; 213.2). The inpatients have a significantly inferior survival (median=145 days; CI 95%=135 – 155) than the outpatients (median=221 days; CI 95%=199 – 243). Patients on stage IV have a significantly inferior survival (median=176 days; CI 95%=127 - 225) than patients on stage IIIb (median=221; CI 95%= 147 - 295). Patients with hepatic metastasis have a significantly inferior survival (median=115 days; CI 95%=74 - 155) than patients without hepatic metastasis (median=199 days; CI 95%=174 - 224). Patients with pleural metastasis have a significantly superior survival (median=386 days; CI 95%=129 – 643) than patients without pleural metastasis (median=174; CI 95%=133 – 214). There was no meaningful statistical difference between survival curves of different nutritional stages (log rank test; p=0,652). Bedridden patients have a significantly inferior survival (median=115 days; CI 95%=87 – 143) than patients with normal physical capacity (median=255 days; CI 95%=202 – 308). The higher the total body depletion, the lower the patient survival (p=0.014). The increase of 1 point in this item of the PG-SGA (IDR=1.70; CI 95%=1.12 – 2.59) may increase risk of death, averagely, by 70%. Karnofsky index was also significantly associated with survival (p=0.016), seeing that for an increase of 10% in the performance scale the risk of death decreased in 23% (IDR=0.77; CI 95%=0.76 – 0.79). The most prevalent symptoms that affect nourishment among the patients were lack of appetite (66.7%), depression (51%) and pain (39.2%) and the least mentioned were mouth pain and vomit (5.9%). Factors that remain statistically associated with death after the adjust by the multivariate analysis by regression of Cox were hepatic metastasis and hospital internment as risk factors for death, and pleural metastasis and better Karnofsky index as protective factors. Conclusion: The survival of eutrophic patients was mildly superior to that of patients in nutritional risk and to that of severely malnourished patients. The higher the total body depletion and the worse the Karnofsky index, the higher the risk of death. No significant statistical association was found between Hemogram, Serum Albumin, and Total Lymphocyte Count (TLC) with the nutritional status of the population in study and survival.

Le système lymphatique a pour rôle principal de drainer les fluides tissulaires et de participer à la surveillance immune. La lymphangiogenèse est principalement activée par deux facteurs : les vascular endothelial growth factors-C et -D (VEGF-C et VEGF-D). Ces facteurs participent à la progression métastatique à la fois en stimulant la prolifération des vaisseaux lymphatiques dans la tumeur et en périphérie et en provoquant la dilatation des vaisseaux collecteurs facilitant ainsi le passage des cellules tumorales de la tumeur primaire aux ganglions lymphatiques. Lors de son développement, une tumeur est soumise à de nombreux stress cellulaires tels que l'hypoxie, l'inflammation ou la déprivation en nutriments. Ma thèse a donc été consacrée à l'analyse des régulations moléculaires permettant une surexpression des facteurs lymphangiogéniques en condition de stress cellulaire. Lors d'un premier projet de recherche j'ai démontré que l'hypoxie réduit la transcription de VEGF-C ainsi que sa traduction coiffe-dépendante mais activait une traduction médiée par un site interne d'entrée du ribosome (IRES). De plus, cette activation de la synthèse de VEGF-C en hypoxie est indépendante de HIF-1a et stimule la lymphangiogenèse dans les tumeurs ainsi que dans les ganglions lymphatiques participant ainsi à la dissémination tumorale. Dans un second temps, j'ai démontré que le VEGF-D possédait lui aussi un IRES activé par un choc thermique. L'activité de l'IRES du VEGF-D est régulée par la localisation subcellulaire d'un facteur trans-activateur de l'IRES (ITAF) : la nucléoline. Cette protéine est exportée du noyau vers le cytoplasme lors d'un choc thermique. Ce processus est inhibé par un traitement avec un anti-inflammatoire non stéroïdien ce qui supprime l'activation de la traduction IRES dépendante du VEGF-D. Enfin lors d'un troisième projet, j'ai travaillé sur le lymphœdème secondaire, une pathologie caractérisée par une destruction du système lymphatique où la lymphangiogenèse doit être stimulée. J'ai évalué l'impact de l'hormonothérapie, principal traitement du cancer du sein, sur l'expression du VEGF-C et -D. J'ai démontré que l'estradiol stimulait l'expression de ces facteurs alors que l'hormonothérapie diminue leur synthèse et provoque une destruction de l'endothélium lymphatique. En conclusion, mes trois projets de thèse m'ont permis d'étudier les régulations moléculaires des deux facteurs lymphangiogéniques principaux dans des pathologies provoquées par une lymphangiogenèse excessive ou insuffisante. / Tumor lymphangiogenesis promotes lymph node metastasis using two mechanisms consisting of lymphatic vessels proliferation and dilatation to facilitate tumor spread. Lymphangiogenesis is mainly promoted by two growth factors: the vascular endothelial growth factor C and D. My PhD was focused on the molecular regulations inducing a tumoral over-expression of these two factors during stress. I demonstrated that hypoxia reduced VEGF-C transcription and cap-dependent translation while activating an Internal Ribosome Entry Site (IRES)-dependent mechanism of translation. This upregulation of VEGF-C in hypoxia is independent of HIF-1a; and stimulates lymphangiogenesis in tumors and lymph nodes and contribute to lymphatic metastasis. Then, I have discovered that VEGF-D had an IRES selectively activated by heat shock. VEGF-D IRES is regulated by a subcellular relocalization of an ITAF, the nucleolin. This molecular process is reversed by non-steroidal anti-inflammatory drugs (NSAID) that decrease VEGF-D IRES translation initiation by targeting the nucleolin. Finally, I worked on secondary lymphedema, a pathology that is characterized by a disruption of the lymphatic network where lymphangiogenesis thus need to be restored. In this condition, I have evaluated the impact of hormone therapy, the main treatment for breast cancer, on the regulation of lymphangiogenic factors. I found that estradiol stimulates the expression of VEGF-C and-D contrary to the hormone therapy that inhibits VEGF-C and -D and thus mediates a disruption of the lymphatic endothelium. Taken together, my PhD projects have allowed me to study the regulations of VEGF-C and -D in pathologies mediated either by an excessive or an insufficient lymphangiogenesis.

La prise en charge contemporaine du cancer repose de plus en plus sur le concept de médecine dite personnalisée qui permet, après identification des caractéristiques propres de la tumeur, d'envisager une thérapeutique adaptée à chaque patient. La meilleure connaissance de la biologie des cancers et la mise en évidence de nouvelles cibles thérapeutiques ont ainsi abouti au développement des thérapies ciblées qui vont agir sur une cible cellulaire, généralement surexprimée au niveau d'une tumeur, leur conférant une plus grande spécificité d'action tumorale et un profil d'effets indésirables moins marqué par rapport à la thérapie cytotoxique conventionnelle. Malgré les données d'efficacité clinique, rapportées sur les stratégies thérapeutiques intégrant des thérapies ciblées, on ne dispose pas, malheureusement, des études pharmaco-économiques en cancérologie surtout que ces médicaments se caractérisent par leurs coûts très élevés (exemple: bevacizumab 1000 euros/400 mg ; cetuximab 964 euros/500 mg) et doivent donc être également appréhendés dans le cadre d'études dites en condition de vie réelle. L'objectif de ce projet de recherche est donc de proposer une étude pharmaco- économique sur l'utilisation des thérapies ciblées en cancérologie. Compte tenu du large champ d'investigation, deux affections tumorales distinctes seront considérées : le cancer colorectal métastatique et le glioblastome.Le cancer colorectal fera l'objet d'une étude conduite spécifiquement au Liban et le glioblastome d'une étude en France. L'originalité de cette thèse, conduite dans le cadre d'une coopération franco-libanaise, sera également d'évaluer les impacts médico-économiques de ces nouvelles stratégies selon des perspectives d'organismes payeurs très différentes, ce qui permettra d'appréhender les éventuelles problématiques d'accès aux soins / Although cancer incidence and prevalence are increasing at an alarming rate, progress in the treatment has been slow and treatment benefits are measured in weeks to months. Following the progress in the diagnostic tools and early detection biomarkers, a number of cancer types can be detected before pathological symptoms develops and this is of great significance because individual specific treatment regimens can be designed based on the presence and stage of cancer. In the past decade, there have been considerable improvements in the way that tumors are characterized and knowledge of cancer, at the molecular level has therefore increased greatly and has shift towards the use of targeted cancer therapies that display greater sensitivity and specificity for tumor cells by blocking the activity of the molecule in the host microenvironment that supports tumor growth or inhibiting the activity of protein tyrosine kinases or signaling pathways. Unfortunately, many pharmaco-economic studies are lacking today in the oncologic field. In addition, targeted therapies are very expensive (bevacizumab costs 100 euros/400mg and cetuximab 964 euros/500 mg) and should, therefore, be analyzed in real life conditions. That’s why the main objective of this research-project is to find a pharmaco-economic method on the use of the targeted therapies in oncology. Given the wide scope of investigations, 2 different tumor pathologies will be discussed: metastatic colorectal cancer and glioblastoma. The choice between these 2 cancer types takes place in the growing role of the targeted therapies in the effectiveness and survival of patients as well as their relative costs, especially that they have in common “bevacizumab” targeting the tumor vasculature

Previous studies have shown that the upregulation of CD151 and β1 is associated with poor prognosis in many cancers such as breast cancer. Studies have provided evidence that these proteins are associated with the adhesion and migration of tumor cells. In this study, a microfluidic flow chamber was utilized to determine how CD151 and β1 affected the firm and initial adhesion of metastatic breast cancer cells to a planar endothelial monolayer under shear stress. This system mimicked the adhesion of metastatic breast cancer cells to the endothelial cells of the circulatory system. CD151 and β1 increased the firm adhesion of metastatic breast cancer cells onto an endothelial monolayer when subjected to high shear stresses. CD151 and β1 increased the initial adhesion of metastatic breast cancer cells onto an endothelial monolayer. A transwell assay was utilized to determine how CD151 and β1 affected random migration through different matrixes and random transendothelial migration. CD151 and β1 decreased the random migration of metastatic breast cancer cells through matrices. Additionally, background information is provided related to the metastatic cascade, how it can be modeled with microfluidics, and how CD151 and β1 have been known to effect cancer and metastasis.

Introdução: O câncer de pulmão é um dos tipos mais comuns de câncer, sendo ele responsável pelo maior número de mortes por neoplasias no mundo. Pacientes com câncer de pulmão apresentam alta prevalência de desnutrição, que está relacionada com piora do prognóstico. A desnutrição ocorre em 60% dos pacientes no momento do diagnóstico, levando a um mau prognóstico, independente do estágio do tumor. Avaliação nutricional de pacientes com câncer deve ser realizada à luz de qualquer proposta terapêutica, ainda que esta seja paliativa. Neste sentido, o método validado de Avaliação Subjetiva Global Produzida pelo Próprio Paciente (ASG-PPP) e adaptado por Ottery et al para pacientes oncológicos a partir da Avaliação Subjetiva Global, ajuda a identificar precocemente os pacientes que estão em risco nutricional. A nutrição é fundamental nestes pacientes em doença avançada, pois fornece a quantidade adequada de nutrientes, tem um importante papel psicológico, social, espiritual e cultural, além de ajudar a manter um sentido de autonomia e bem-estar. Objetivo: Relacionar a Capacidade Funcional através do índice de Karnofsky e o estado nutricional, sinais e sintomas avaliados pelo método subjetivo (ASG-PPP), com a sobrevida de pacientes com neoplasia pulmonar mestastática. Pacientes e métodos: Trata-se de um estudo transversal, constituído por 51 pacientes, ambos os sexos, portadores de neoplasia pulmonar com doença metastática em estadiamento IIIb e IV, atendidos em nível ambulatorial e hospitalar no Hospital Santa Rita, Complexo Hospitalar Santa Casa de Porto Alegre. A sobrevida dos pacientes foi verificada ao final do estudo, através de revisão de prontuários e contato telefônico com o responsável pelo paciente. Resultados: Os pacientes apresentaram-se com idade 61,84±10,83 anos (40 a 82 anos); 34 (66,7%) eram gênero masculino e 17 (33,3%) do feminino. A mediana do tempo de sobrevida foi de 186 dias (IC 95%: 158,8; 213,2). Os pacientes internados tiveram sobrevida significativamente inferior (mediana=145 dias (IC 95%=135 - 155) aos que foram atendidos ambulatorialmente (mediana=221 dias (IC 95%=199 - 243). Os pacientes em estágio IV tiveram sobrevida significativamente inferior (mediana=176 dias; IC 95%=127 - 225) àqueles em estágio IIIb (mediana=221; IC 95%=147 - 295). Os pacientes com metástase hepática tiveram sobrevida significativamente inferior (mediana=115 dias; IC 95%=74 - 155) aos sem metástase hepática (mediana=199; IC 95%=174 - 224). Os pacientes com metástase pleural apresentaram sobrevida significativamente superior (mediana=386 dias; IC 95%=129 - 643) aos sem metástase pleural (mediana=174; IC 95%=133 - 214). Não houve diferença estatisticamente significativa entre as curvas de sobrevida dos diferentes estágios nutricionais (teste de log rank; p=0,651). Os acamados tiveram sobrevida significativamente inferior (mediana=115 dias; IC 95%=87 - 143) que os com capacidade física normal (mediana=255 dias; IC 95%=202 - 308). Quanto maior a depleção corporal total, pior a sobrevida do paciente (p=0,014). O aumento de 1 ponto neste item da ASG-PPP (RDI=1,70; IC 95%=1,12 – 2,59) pode aumentar o risco de óbito, em média 70%. O índice de Karnofsky também se associou significativamente com a sobrevida (p=0,016), sendo que para um aumento de 10% na escala de nível de desempenho, o risco de óbito diminuiu em 23% (RDI=0,77; IC 95%=0,76 – 0,79). Os sintomas mais prevalentes que afetaram a alimentação entre os pacientes foram a falta de apetite (66,7%), depressão (51%) e dor (39,2%) e os menos relatados foram dor na boca e vômitos (5,9%). Os fatores que permaneceram associados estatisticamente com o óbito após o ajuste pela análise multivariada de Regressão de Cox foram metástase hepática e internação hospitalar como fatores de risco para óbito e metástase pleural e melhor índice de Karnofsky como fatores protetores. Conclusão: A sobrevida dos pacientes eutróficos foi levemente superior a dos pacientes em risco nutricional e a dos severamente desnutridos. Quanto maior a depleção corporal total e pior o índice de Karnofsky maior o risco de óbito. Não foi encontrada uma associação estatisticamente significativa entre os valores de Hemograma, Albumina Sérica e Contagem Total de Linfócitos (CTL) com o estado nutricional da população em estudo e a sobrevida. / Introduction: Lung cancer is one of the most common cancers in the world, and it is today the most lethal of the neoplastic diseases. Patients with lung cancer show high prevalence of malnutrition, the latter being related to worsening of the prognosis. Malnutrition occurs in 60% of the patients at the moment of diagnosis, leading to a bad prognosis, regardless of the tumor stage. Nutritional evaluation of patients with cancer should be conducted in the light of a therapeutic proposal, even if it’s palliative. In this way, the validated method Scored Patient-Generated Subjective Global Assessment (PG-SGA), adapted by Ottery et al for oncologic patients from the questionnaire of Subjective Global Assessment, helps to precociously identify patients that are in nutritional risk. Nutrition is fundamental in these patients with advanced disease for it supplies the adequate amount of nutrients, performs an important psychological, spiritual and cultural role, besides helping to build a sense of autonomy and well-being. Objective: to relate Functional Capacity through the Karnofsky Index and the nutritional state, signs and symptoms evaluated by the subjective method (PG-SGA), to the survival of patients with metastatic pulmonary neoplasia. Patients and methods: It’s a transversal study, constituted by 51 patients, both genders, carriers of pulmonary neoplasia with metastatic disease on stages IIIb and IV, both in and outpatients at Santa Rita Hospital, Complexo Hospitalar Santa Casa de Porto Alegre. The survival of the patients in the study was assessed at the end of the study, through review of the chart and phone contact with the patient’s keeper. Results: The average age is 61.84 (± 10.83 varying between 40 and 82), being 34 (66.7%) male and 17 (33.3%) female. The median of survival is 186 days (CI 95%; 158.8; 213.2). The inpatients have a significantly inferior survival (median=145 days; CI 95%=135 – 155) than the outpatients (median=221 days; CI 95%=199 – 243). Patients on stage IV have a significantly inferior survival (median=176 days; CI 95%=127 - 225) than patients on stage IIIb (median=221; CI 95%= 147 - 295). Patients with hepatic metastasis have a significantly inferior survival (median=115 days; CI 95%=74 - 155) than patients without hepatic metastasis (median=199 days; CI 95%=174 - 224). Patients with pleural metastasis have a significantly superior survival (median=386 days; CI 95%=129 – 643) than patients without pleural metastasis (median=174; CI 95%=133 – 214). There was no meaningful statistical difference between survival curves of different nutritional stages (log rank test; p=0,652). Bedridden patients have a significantly inferior survival (median=115 days; CI 95%=87 – 143) than patients with normal physical capacity (median=255 days; CI 95%=202 – 308). The higher the total body depletion, the lower the patient survival (p=0.014). The increase of 1 point in this item of the PG-SGA (IDR=1.70; CI 95%=1.12 – 2.59) may increase risk of death, averagely, by 70%. Karnofsky index was also significantly associated with survival (p=0.016), seeing that for an increase of 10% in the performance scale the risk of death decreased in 23% (IDR=0.77; CI 95%=0.76 – 0.79). The most prevalent symptoms that affect nourishment among the patients were lack of appetite (66.7%), depression (51%) and pain (39.2%) and the least mentioned were mouth pain and vomit (5.9%). Factors that remain statistically associated with death after the adjust by the multivariate analysis by regression of Cox were hepatic metastasis and hospital internment as risk factors for death, and pleural metastasis and better Karnofsky index as protective factors. Conclusion: The survival of eutrophic patients was mildly superior to that of patients in nutritional risk and to that of severely malnourished patients. The higher the total body depletion and the worse the Karnofsky index, the higher the risk of death. No significant statistical association was found between Hemogram, Serum Albumin, and Total Lymphocyte Count (TLC) with the nutritional status of the population in study and survival.

abstract: Osteosarcoma is the most common bone cancer in children and adolescents. Patients with metastatic osteosarcoma are typically refractory to treatment. Numerous lines of evidence suggest that cytotoxic T-lymphocytes (CTL) limit the development of metastatic osteosarcoma. I have investigated the role of Programmed Death Receptor-1 (PD-1) in limiting the efficacy of immune mediated control of metastatic osteosarcoma. I show that human metastatic, but not primary, osteosarcoma tumors express the ligand for PD-1 (PD-L1) and that tumor infiltrating CTL express PD-1, suggesting this pathway may limit CTL control of metastatic osteosarcoma in patients. PD-L1 is also expressed on the K7M2 osteosarcoma tumor cell line that establishes metastases in mice, and PD-1 is expressed on tumor infiltrating CTL during disease progression. Blockade of PD-1/PD-L1 interactions dramatically improves the function of osteosarcoma-reactive CTL in vitro and in vivo, and results in decreased tumor burden and increased survival in the K7M2 mouse model of metastatic osteosarcoma. My results suggest that blockade of PD-1/PD-L1 interactions in patients with metastatic osteosarcoma should be pursued as a therapeutic strategy. However, PD-1/PD-L1 blockade treated mice still succumb to disease due to selection of PD-L1 mAb resistant tumor cells via up-regulation of other co-inhibitory T cell receptors. Combinational α-CTLA-4 and α-PD-L1 blockade treated mice were able to completely eradicate metastatic osteosarcoma, and generate immunity to disease. These results suggest that blockade of PD-1/PD-L1 interactions in patients with metastatic osteosarcoma, although improves survival, may lead to tumor resistance, requiring combinational immunotherapies to combat and eradicate disease. / Dissertation/Thesis / Doctoral Dissertation Molecular and Cellular Biology 2015

Introdução: O câncer de pulmão é um dos tipos mais comuns de câncer, sendo ele responsável pelo maior número de mortes por neoplasias no mundo. Pacientes com câncer de pulmão apresentam alta prevalência de desnutrição, que está relacionada com piora do prognóstico. A desnutrição ocorre em 60% dos pacientes no momento do diagnóstico, levando a um mau prognóstico, independente do estágio do tumor. Avaliação nutricional de pacientes com câncer deve ser realizada à luz de qualquer proposta terapêutica, ainda que esta seja paliativa. Neste sentido, o método validado de Avaliação Subjetiva Global Produzida pelo Próprio Paciente (ASG-PPP) e adaptado por Ottery et al para pacientes oncológicos a partir da Avaliação Subjetiva Global, ajuda a identificar precocemente os pacientes que estão em risco nutricional. A nutrição é fundamental nestes pacientes em doença avançada, pois fornece a quantidade adequada de nutrientes, tem um importante papel psicológico, social, espiritual e cultural, além de ajudar a manter um sentido de autonomia e bem-estar. Objetivo: Relacionar a Capacidade Funcional através do índice de Karnofsky e o estado nutricional, sinais e sintomas avaliados pelo método subjetivo (ASG-PPP), com a sobrevida de pacientes com neoplasia pulmonar mestastática. Pacientes e métodos: Trata-se de um estudo transversal, constituído por 51 pacientes, ambos os sexos, portadores de neoplasia pulmonar com doença metastática em estadiamento IIIb e IV, atendidos em nível ambulatorial e hospitalar no Hospital Santa Rita, Complexo Hospitalar Santa Casa de Porto Alegre. A sobrevida dos pacientes foi verificada ao final do estudo, através de revisão de prontuários e contato telefônico com o responsável pelo paciente. Resultados: Os pacientes apresentaram-se com idade 61,84±10,83 anos (40 a 82 anos); 34 (66,7%) eram gênero masculino e 17 (33,3%) do feminino. A mediana do tempo de sobrevida foi de 186 dias (IC 95%: 158,8; 213,2). Os pacientes internados tiveram sobrevida significativamente inferior (mediana=145 dias (IC 95%=135 - 155) aos que foram atendidos ambulatorialmente (mediana=221 dias (IC 95%=199 - 243). Os pacientes em estágio IV tiveram sobrevida significativamente inferior (mediana=176 dias; IC 95%=127 - 225) àqueles em estágio IIIb (mediana=221; IC 95%=147 - 295). Os pacientes com metástase hepática tiveram sobrevida significativamente inferior (mediana=115 dias; IC 95%=74 - 155) aos sem metástase hepática (mediana=199; IC 95%=174 - 224). Os pacientes com metástase pleural apresentaram sobrevida significativamente superior (mediana=386 dias; IC 95%=129 - 643) aos sem metástase pleural (mediana=174; IC 95%=133 - 214). Não houve diferença estatisticamente significativa entre as curvas de sobrevida dos diferentes estágios nutricionais (teste de log rank; p=0,651). Os acamados tiveram sobrevida significativamente inferior (mediana=115 dias; IC 95%=87 - 143) que os com capacidade física normal (mediana=255 dias; IC 95%=202 - 308). Quanto maior a depleção corporal total, pior a sobrevida do paciente (p=0,014). O aumento de 1 ponto neste item da ASG-PPP (RDI=1,70; IC 95%=1,12 – 2,59) pode aumentar o risco de óbito, em média 70%. O índice de Karnofsky também se associou significativamente com a sobrevida (p=0,016), sendo que para um aumento de 10% na escala de nível de desempenho, o risco de óbito diminuiu em 23% (RDI=0,77; IC 95%=0,76 – 0,79). Os sintomas mais prevalentes que afetaram a alimentação entre os pacientes foram a falta de apetite (66,7%), depressão (51%) e dor (39,2%) e os menos relatados foram dor na boca e vômitos (5,9%). Os fatores que permaneceram associados estatisticamente com o óbito após o ajuste pela análise multivariada de Regressão de Cox foram metástase hepática e internação hospitalar como fatores de risco para óbito e metástase pleural e melhor índice de Karnofsky como fatores protetores. Conclusão: A sobrevida dos pacientes eutróficos foi levemente superior a dos pacientes em risco nutricional e a dos severamente desnutridos. Quanto maior a depleção corporal total e pior o índice de Karnofsky maior o risco de óbito. Não foi encontrada uma associação estatisticamente significativa entre os valores de Hemograma, Albumina Sérica e Contagem Total de Linfócitos (CTL) com o estado nutricional da população em estudo e a sobrevida. / Introduction: Lung cancer is one of the most common cancers in the world, and it is today the most lethal of the neoplastic diseases. Patients with lung cancer show high prevalence of malnutrition, the latter being related to worsening of the prognosis. Malnutrition occurs in 60% of the patients at the moment of diagnosis, leading to a bad prognosis, regardless of the tumor stage. Nutritional evaluation of patients with cancer should be conducted in the light of a therapeutic proposal, even if it’s palliative. In this way, the validated method Scored Patient-Generated Subjective Global Assessment (PG-SGA), adapted by Ottery et al for oncologic patients from the questionnaire of Subjective Global Assessment, helps to precociously identify patients that are in nutritional risk. Nutrition is fundamental in these patients with advanced disease for it supplies the adequate amount of nutrients, performs an important psychological, spiritual and cultural role, besides helping to build a sense of autonomy and well-being. Objective: to relate Functional Capacity through the Karnofsky Index and the nutritional state, signs and symptoms evaluated by the subjective method (PG-SGA), to the survival of patients with metastatic pulmonary neoplasia. Patients and methods: It’s a transversal study, constituted by 51 patients, both genders, carriers of pulmonary neoplasia with metastatic disease on stages IIIb and IV, both in and outpatients at Santa Rita Hospital, Complexo Hospitalar Santa Casa de Porto Alegre. The survival of the patients in the study was assessed at the end of the study, through review of the chart and phone contact with the patient’s keeper. Results: The average age is 61.84 (± 10.83 varying between 40 and 82), being 34 (66.7%) male and 17 (33.3%) female. The median of survival is 186 days (CI 95%; 158.8; 213.2). The inpatients have a significantly inferior survival (median=145 days; CI 95%=135 – 155) than the outpatients (median=221 days; CI 95%=199 – 243). Patients on stage IV have a significantly inferior survival (median=176 days; CI 95%=127 - 225) than patients on stage IIIb (median=221; CI 95%= 147 - 295). Patients with hepatic metastasis have a significantly inferior survival (median=115 days; CI 95%=74 - 155) than patients without hepatic metastasis (median=199 days; CI 95%=174 - 224). Patients with pleural metastasis have a significantly superior survival (median=386 days; CI 95%=129 – 643) than patients without pleural metastasis (median=174; CI 95%=133 – 214). There was no meaningful statistical difference between survival curves of different nutritional stages (log rank test; p=0,652). Bedridden patients have a significantly inferior survival (median=115 days; CI 95%=87 – 143) than patients with normal physical capacity (median=255 days; CI 95%=202 – 308). The higher the total body depletion, the lower the patient survival (p=0.014). The increase of 1 point in this item of the PG-SGA (IDR=1.70; CI 95%=1.12 – 2.59) may increase risk of death, averagely, by 70%. Karnofsky index was also significantly associated with survival (p=0.016), seeing that for an increase of 10% in the performance scale the risk of death decreased in 23% (IDR=0.77; CI 95%=0.76 – 0.79). The most prevalent symptoms that affect nourishment among the patients were lack of appetite (66.7%), depression (51%) and pain (39.2%) and the least mentioned were mouth pain and vomit (5.9%). Factors that remain statistically associated with death after the adjust by the multivariate analysis by regression of Cox were hepatic metastasis and hospital internment as risk factors for death, and pleural metastasis and better Karnofsky index as protective factors. Conclusion: The survival of eutrophic patients was mildly superior to that of patients in nutritional risk and to that of severely malnourished patients. The higher the total body depletion and the worse the Karnofsky index, the higher the risk of death. No significant statistical association was found between Hemogram, Serum Albumin, and Total Lymphocyte Count (TLC) with the nutritional status of the population in study and survival.

1. Abstract Pheochromocytoma/ paraganglioma (FEO/PGL) may be developed on the basis of an inherited genetic mutation of different genes. They are associated with a high risk of developing of secondary hypertension, organ damage and metastatic disease that can be fatal. The aim was to focus on the possibility of genetic testing in patients with FEO/PGL, especially in patients with malignant tumors. The issue FEO/PGL, however, concerns not only the examination and assessment of risks arising therefrom, as well as other therapies and monitoring, including appropriate recommendations for clinical practice. We demonstrated a 20% incidence of cardiovascular (CV) complications before determining the final diagnosis of FEO/PGL, mainly arrhythmic, followed by complications of myocardial ischemia and accentuate atherosclerosis. Elevated levels of vitamin C and decreased levels of malondialdehyde (MDA) following the successful removal of the tumor demonstrated reduction of oxidative stress postoperatively. We found that early postoperative testing of levels of plasma metanephrines to confirm the success of surgical removal of FEO/PGL is already possible, since there was no significant correlation between plasma levels of metanephrines and postoperative examination interval. Distribution of frequency of metastatic...

Hintergrund: Ziel der Studie war der Vergleich von Patienten mit duktalem Pankreaskarzinom (PDAC) im progressionsfreien und Gesamtüberleben sowie im Rezidivmuster in Abhängigkeit einer Resektion der Vena portae oder der Vena mesenterica superior (PV/SMV). Methoden: Es wurde eine retrospektive Analyse durchgeführt. Hierbei wurden Patienten betrachtet, die zwischen 2005 und 2015 eine pyloruserhaltende partielle Pankreatoduodenektomie (PPPD), eine klassische Pankreatoduodenektomie (kPD) oder eine totale Pankreatektomie (TP) erhielten. Diese wurden in drei Gruppen eingeteilt. Die P+I+- Gruppe bestand aus Patienten mit Venenresektion (P+), bei denen eine pathohistologische Infiltration der PV oder SMV vorlag (I+). Fand sich bei durchgeführter Venenresektion keine portalvenöse Infiltration (I-), wurden die Patienten der P+I--Gruppe zugeordnet. Als Kontrollgruppe galten Patienten ohne Venenresektion (P-I-), welche zu denen der P+I+- Gruppe gematcht wurden. Die statistischen Analysen wurden mit dem R Softwarepaket durchgeführt. Das Signifikanzlevel wurde für alle Berechnungen auf α = 0,05 festgelegt. Ergebnisse: Insgesamt wurden 179 Patienten eingeschlossen. 113 erhielten eine portalvenöse Resektion. Davon hatten 36 (31,9%) eine pathohistologische Lumeninfiltration (P+I+), bei 77 (68,1%) lag dagegen keine Infiltration vor (P+I-). 66 Patienten ohne Venenresektion wurden zu den Patienten der P+I+-Gruppe gematcht (P-I-). Zwischen den drei Gruppen waren die meisten pathohistologischen Parameter vergleichbar. 17 Patienten (9,5%) wurden neoadjuvant therapiert, davon erhielten 16 eine Venenresektion (P+). Für das Gesamtüberleben konnten signifikante Unterschiede nachgewiesen werden (11,9 Monate [P+I+] vs. 16,1 Monate [P+I-] vs. 20,1 Monate [P-I-]; p=0,01). In der univariaten Überlebensanalyse konnte für den erhöhten präoperativen CA19-9 Wert, den Resektionsstatus (R), den Lymphknotenstatus (N), das Lymphknotenverhältnis (LNR), die mikroskopische Veneninvasion (V) sowie die pathohistologisch gesicherte Infiltration der PV/SMV ein negativer Einfluss nachgewiesen werden. In der multivariaten Analyse blieb die wahre Infiltration der PV/SMV als einziger signifikanter negativer Einflussfaktor auf das Gesamtüberleben erhalten (p=0,014). Die Inzidenz an Fernmetastasen war in der P+I+- Gruppe signifikant erhöht (75% [P+I+] vs. 45,8% [P+I-] vs. 54,7% [P-I-], p=0,01). Für ein Lokalrezidiv fanden sich dagegen keine Häufigkeitsunterschiede zwischen den Gruppen (p=0,96). Das mediane progressionsfreie Überleben war für Patienten der P+I+-Gruppe signifikant verkürzt (7,4 Monate [P+I+] vs. 10,9 Monate [P+I-] vs. 11,6 Monate [P-I-]; p=0,02). Die Lumeninfiltration der PV/SMV, die mikroskopische Veneninvasion (V), der präoperative CA19-9 Wert sowie der Differenzierungsgrad (G) waren negative Einflussfaktoren auf das progressionsfreie Überleben. In der multivariaten Analyse blieben die pathohistologisch gesicherte Infiltration sowie das Grading als negative unabhängige Einflussfaktoren nachweisbar. In 25% der Fälle manifestierte sich das Rezidiv initial in der Leber. Schlussfolgerung: Die pathohistologisch gesicherte Infiltration der PV/SMV ist ein unabhängiger Risikofaktor für das progressionsfreie und das Gesamtüberleben. Die Inzidenz an Fernmetastasen ist für die Patienten der P+I+-Gruppe erhöht. Eine potentiell kurative venöse Resektion kann den Einfluss der aggressiven Tumorbiologie und des fortgeschrittenen Krankheitsbildes nicht vollständig kompensieren. / Background. The present study aims to evaluate the longterm outcome and metastatatic pattern of patients who underwent an operation for pancreatic ductal adenocarcinoma (PDAC) with portal or superior mesenteric vein (PV/SMV) resection. Methods. Patients who underwent a pylorus preserving pancreaticoduodenectomy (PPPD), Whipple procedure (kPD) or total pancreatoduodenectomy (TP) between 2005 and 2015 were retrospectively analyzed. The patients were categorized in three subgroups. Those whom received a vein resection with pathohistological tumor invasion of the PV/SMV (P+I+) those at whom underwent vein resection but without pathohistological tumor invasion (P+I-) and lastly a third group (P-I-) matched to the P+I+ included patients without vein resection. Statistical analysis was performed using the R software package. The significance level for all calculations was set at α = 0.05. Results. The study cohort included 179 patients, 113 of whom underwent simultaneous PV/SMV resection. 36 patients (31,9%) had pathohistological tumor infiltration (P+I+), 77 (68,1%) did not (P+I-). 66 patients without vein resection (P-I-) were balanced by the P+I+ group. Most of pathohistological tumor characteristics were comparable between groups. 17 patients (9.5%) received neoadjuvant therapy, 16 of them were in vein resection group (P+). The study revealed differences in overall median survival (11.9 months [P+I+] vs. 16.1 months [P+I-] vs. 20.1 months [P-I-]; p=0.01). Univariate survival analysis shown negative consequences for CA19-9, resection margin (R), status of nodal metastasis (N), lymph node ratio (LNR), microvascular vein invasion (V) and true invasion of the PV/SMV. Multivariate survival analysis identified true invasion of the PV/SMV as the only significant, negative prognostic factor (p= 0.01). Whereas the incidence of local tumor recurrence was comparable (p=0.96), the proportion of patients with distant metastasis showed significant differences (75% [P+I+] vs. 45.8% [P+I-] vs 54.7% [P-I-]; p=0.01). The median time to progression were significantly shorter if the PV/SMV was infiltrated (7,4 months [P+I+] vs. 10,9 months [P+I-] vs. 11,6 months [P-I-]; p=0.02). Univariate progression analysis revealed significances for true invasion of the PV/SMV, microvascular vein invasion (V), CA19-9 and histologic classification (G). Multivariate progression analysis detected pathohistological invasion of the PV/SMV and histologic classification (G) as independent factors. Initial liver metastasis occurred in 25% of the patients. Conclusions. Pathohistological invasion of the PV/SMV is an independent risk factor for overall and progression free survival. Patients of P+I+-group had a higher incidence of distant metastasis, local progression is comparable. Even radical and complete resection cannot completely compensate for aggressive tumor biology and advanced disease. Modifiziert nach Mierke et al., 2016