Introdução: O Adenocarcinoma ductal pancreático é um tumor bastante agressivo que apresenta uma alta de letalidade e, para o qual, poucas opções terapêuticas estão disponíveis. Isto pode ser parcialmente explicado pela complexidade derivada de múltiplas aberrações genéticas e da população celular mista presente em um tumor pancreático, o que também pode explicar o curso clínico heterogêneo observado na prática diária. Ultimamente, pesquisas científicas têm contribuído para ampliar o conhecimento sobre o impacto das alterações epigenéticas no desenvolvimento de múltiplos tipos de câncer, porém no pâncreas essas alterações ainda são incertas e, por isso, foco de investigação. A desregulação epigenética parece estar envolvida no ciclo celular da célula tumoral, incluindo o crescimento celular, diferenciação, progressão tumoral e morte celular, e a acetilação das histonas é um importante mecanismo que regula a transcrição de genes envolvidos nesses processos. Padrões globais de modificações das histonas foram recentemente apontados como preditores de desfecho em pacientes com câncer, mas poucos estudos têm sido realizados nesta área, inclusive em Adenocarcinoma ductal pancreático (ADP). Objetivos: O presente estudo foi desenvolvido a fim de investigar o padrão de modificação de acetilação das histonas em adenocarcinoma pancreático, através da análise imunohistoquímica. Materiais e métodos: Uma análise clinicopatológica retrospectiva foi realizada em 119 pacientes diagnosticados com câncer de pâncreas entre os anos de 2005 e 2011, e realizado estudo imunohistoquímico com os anticorpos contra H4K12ac, H3K9ac e H3K18ac. Marcação nuclear positiva para cada histona foi medida quanto à intensidade e expressão, sendo classificadas em grupos de baixa ou de alta intensidade/expressão. Os resultados foram analisados em relação aos parâmetros clinicopatológicos de cada paciente. Resultados: Houve uma relação positiva entre diferenciação tumoral e alta expressão de H4K12ac (P <0,05), bem como a intensidade forte dos três marcadores correlacionou-se positivamente com o estágio do tumor (P <0,01). Análise univariada mostrou pior sobrevida em pacientes com níveis elevados de expressão de H4K12ac (p = 0,038) e H3K18Ac (P = 0,033). Modelo de risco proporcional de Cox revelou o efeito prognóstico independente de níveis elevados de H4K12ac H3K18ac (taxas de risco de 1,6 e 1,7, respectivamente, p <0,05), especialmente para pacientes em estágios iniciais. Sugerimos como hipótese que as modificações na acetilação das histonas H4K12 e H3K18 podem ser consideradas fatores prognósticos importantes para o câncer de pâncreas, embora o mecanismo envolvido necessite de mais investigação. Aumentando a compreensão e o conhecimento sobre o padrão de acetilação das histonas, poder-se-ão finalmente gerar novas idéias para um diagnóstico molecular racional e novas abordagens terapêuticas. / Introduction: Ductal pancreatic adenocarcinoma (DPAs) is a highly aggressive tumor, with a high letality rate, for which few therapeutic options are available. This may be partially explained by the notorious complexity derived from the multiple genetic aberrations and mixed cellular population present in a pancreatic tumor, which can also explain the heterogeneous clinical course observed in daily practice. Lately, there is an increase in the literature about the impact of epigenetic changes on the development of several cancer, however in the pancreas these changes are still uncertain. Epigenetic deregulation may be involved in tumor cell biology, including cell growth, differentiation, tumor progression and cell death, and histone acetylation is a major mechanism that regulates gene transcription. Patterns of global histone modifications have been recently suggested as outcome predictors in cancer patients, but few studies have been conducted on pancreatic ductal adenocarcinomas. Objectives: This study was designed to investigate the predictive value of histone acetylation modifications on pancreatic cancer. Material and methods: A retrospective clinicopathologic analysis was undertaken in 119 patients diagnosed with PDAC between 2005 and 2011, and immunohistochemistry performed with antibodies against H4K12ac, H3K9ac and H3K18ac. Positive nuclear staining for each histone was measured as the intensity and expression, being classified into low or high-staining groups. Results were analyzed in relation to patients’ clinicopathologic parameters. Results: There was a positive relationship between tumor differentiation and H4K12ac high scores (P<0.05) and staining of the three markers correlated positively with tumor stage (P<0.01). Univariate analysis showed worse survival in patients with high detection levels of H4K12ac (p=0.038) and H3K18Ac (P=0.033). A backwards Cox proportional hazards model revealed the independent prognostic effect of high H4K12ac and H3K18ac levels (hazard ratios of 1.6 and 1.7 respectively, p<0.05), especially for patients at early stages. We hypothesize that acetylation of H4K12 and H3K18 may be considered valuable prognostic factors for pancreatic cancer, although the mechanism involved needs further investigation. Increasing insights into histone acetylation modifications can ultimately generate new ideas for rationally and molecularly based diagnostic and therapeutic approaches.

Introdução: a superexpressão de HER-2 tem correlação com maior agressividade em carcinomas de mama e estômago, e a sua detecção já foi incorporada como rotina na análise destas neoplasias. Critérios ideais para avaliação do HER-2 em câncer de pâncreas permanecem incertos. Objetivos: avaliar o status do HER-2 e o seu valor preditivo em adenocarcinoma pancreático. Materiais e métodos: análise clinicopatológica e imuno-histoquímica de 112 pacientes com diagnóstico de câncer de pâncreas com base nos critérios propostos para câncer de mama (HercepTest™) e de estômago (ToGA Trial). Resultados: pelo HercepTest™ 5 (4.5%) casos apresentaram escore 3+, 3 (2.7%) escore 2+ e 104 (92.9%) escores 0/1+. Na análise pelo ToGA Trial, 9 (8.0%) obtiveram escore 3+, 32 (28.6%) escore 2+ e 71 (63.4%) escores 0/1+. Todos os casos positivos pelo HercepTest™ também o foram para o ToGA Trial. Pacientes com hiperexpressão (3+) apresentaram sobrevida média maior que aqueles sem (0 a 2+) tanto pelo HercepTest™ quanto pelo ToGa Trial (43.88 vs. 10.3 meses, p = 0.029 e 40.7 vs. 10.1 meses, p = 0.013, respectivamente). Os demais parâmetros não mostraram correlação com a expressão de HER-2. Conclusão: diferenças na incidência e no significado prognóstico da superexpressão podem ser decorrentes do pequeno tamanho amostral e do uso de dois critérios diferentes de positividade para HER-2. Estes resultados servem com impulso para novas investigações de superexpressão e amplificação do HER-2 utilizando, além da imuno-histoquímica, métodos como FISH e SISH, a fim de se obter mais opções terapêuticas para oferecer aos pacientes, como agentes anti-HER-2. / Introduction: HER-2 overexpression is correlated with aggressiveness in breast and gastric cancers, and its detection has been incorporated as routine in the analysis of these neoplasms. Ideal criteria for evaluation of HER-2 in pancreatic cancer remain unclear. Objectives: to assess the HER-2 status and its predictive value in pancreatic adenocarcinoma. Material and methods: clinicopathologic and immunohistochemical analysis were undertaken in 112 patients with pancreatic cancer using the criteria proposed for breast (HercepTest™) and stomach cancer (ToGA Trial). Results: using HercepTest™ 5 (4.5%) cases had a score of 3+, 3 (2.7%) had a score of 2+ and 104 (92.9%) had scores of 0/1 +. By ToGA Trial, 9 (8.0%) obtained score of 3+, 32 (28.6%) had a score of 2+ score and 71 (63.4%) had scores of 0/1 +. All positive cases by HercepTest™ also went to the ToGA Trial. Patients with overexpression (3 +) showed greater survival than those without (0 to 2 +) by both HercepTest™ and ToGa Trial (43.88 vs. 10.3 months, p = 0.029 and 40.7 vs. 10.1 months, p = 0.013, respectively). Other parameters did not show correlation with the expression of HER-2. Conclusion: differences in incidence and prognostic significance of overexpression may be explained from small sample size and the use of two different criteria of positivity for HER-2. These results serve as an impulse for new investigations of overexpression/amplification of the HER-2 using, besides immunohistochemistry, FISH and SISH methods, in order to get more treatment options to provide patients, as agents anti-HER-2.

Introdução: O Adenocarcinoma ductal pancreático é um tumor bastante agressivo que apresenta uma alta de letalidade e, para o qual, poucas opções terapêuticas estão disponíveis. Isto pode ser parcialmente explicado pela complexidade derivada de múltiplas aberrações genéticas e da população celular mista presente em um tumor pancreático, o que também pode explicar o curso clínico heterogêneo observado na prática diária. Ultimamente, pesquisas científicas têm contribuído para ampliar o conhecimento sobre o impacto das alterações epigenéticas no desenvolvimento de múltiplos tipos de câncer, porém no pâncreas essas alterações ainda são incertas e, por isso, foco de investigação. A desregulação epigenética parece estar envolvida no ciclo celular da célula tumoral, incluindo o crescimento celular, diferenciação, progressão tumoral e morte celular, e a acetilação das histonas é um importante mecanismo que regula a transcrição de genes envolvidos nesses processos. Padrões globais de modificações das histonas foram recentemente apontados como preditores de desfecho em pacientes com câncer, mas poucos estudos têm sido realizados nesta área, inclusive em Adenocarcinoma ductal pancreático (ADP). Objetivos: O presente estudo foi desenvolvido a fim de investigar o padrão de modificação de acetilação das histonas em adenocarcinoma pancreático, através da análise imunohistoquímica. Materiais e métodos: Uma análise clinicopatológica retrospectiva foi realizada em 119 pacientes diagnosticados com câncer de pâncreas entre os anos de 2005 e 2011, e realizado estudo imunohistoquímico com os anticorpos contra H4K12ac, H3K9ac e H3K18ac. Marcação nuclear positiva para cada histona foi medida quanto à intensidade e expressão, sendo classificadas em grupos de baixa ou de alta intensidade/expressão. Os resultados foram analisados em relação aos parâmetros clinicopatológicos de cada paciente. Resultados: Houve uma relação positiva entre diferenciação tumoral e alta expressão de H4K12ac (P <0,05), bem como a intensidade forte dos três marcadores correlacionou-se positivamente com o estágio do tumor (P <0,01). Análise univariada mostrou pior sobrevida em pacientes com níveis elevados de expressão de H4K12ac (p = 0,038) e H3K18Ac (P = 0,033). Modelo de risco proporcional de Cox revelou o efeito prognóstico independente de níveis elevados de H4K12ac H3K18ac (taxas de risco de 1,6 e 1,7, respectivamente, p <0,05), especialmente para pacientes em estágios iniciais. Sugerimos como hipótese que as modificações na acetilação das histonas H4K12 e H3K18 podem ser consideradas fatores prognósticos importantes para o câncer de pâncreas, embora o mecanismo envolvido necessite de mais investigação. Aumentando a compreensão e o conhecimento sobre o padrão de acetilação das histonas, poder-se-ão finalmente gerar novas idéias para um diagnóstico molecular racional e novas abordagens terapêuticas. / Introduction: Ductal pancreatic adenocarcinoma (DPAs) is a highly aggressive tumor, with a high letality rate, for which few therapeutic options are available. This may be partially explained by the notorious complexity derived from the multiple genetic aberrations and mixed cellular population present in a pancreatic tumor, which can also explain the heterogeneous clinical course observed in daily practice. Lately, there is an increase in the literature about the impact of epigenetic changes on the development of several cancer, however in the pancreas these changes are still uncertain. Epigenetic deregulation may be involved in tumor cell biology, including cell growth, differentiation, tumor progression and cell death, and histone acetylation is a major mechanism that regulates gene transcription. Patterns of global histone modifications have been recently suggested as outcome predictors in cancer patients, but few studies have been conducted on pancreatic ductal adenocarcinomas. Objectives: This study was designed to investigate the predictive value of histone acetylation modifications on pancreatic cancer. Material and methods: A retrospective clinicopathologic analysis was undertaken in 119 patients diagnosed with PDAC between 2005 and 2011, and immunohistochemistry performed with antibodies against H4K12ac, H3K9ac and H3K18ac. Positive nuclear staining for each histone was measured as the intensity and expression, being classified into low or high-staining groups. Results were analyzed in relation to patients’ clinicopathologic parameters. Results: There was a positive relationship between tumor differentiation and H4K12ac high scores (P<0.05) and staining of the three markers correlated positively with tumor stage (P<0.01). Univariate analysis showed worse survival in patients with high detection levels of H4K12ac (p=0.038) and H3K18Ac (P=0.033). A backwards Cox proportional hazards model revealed the independent prognostic effect of high H4K12ac and H3K18ac levels (hazard ratios of 1.6 and 1.7 respectively, p<0.05), especially for patients at early stages. We hypothesize that acetylation of H4K12 and H3K18 may be considered valuable prognostic factors for pancreatic cancer, although the mechanism involved needs further investigation. Increasing insights into histone acetylation modifications can ultimately generate new ideas for rationally and molecularly based diagnostic and therapeutic approaches.

Introdução: a superexpressão de HER-2 tem correlação com maior agressividade em carcinomas de mama e estômago, e a sua detecção já foi incorporada como rotina na análise destas neoplasias. Critérios ideais para avaliação do HER-2 em câncer de pâncreas permanecem incertos. Objetivos: avaliar o status do HER-2 e o seu valor preditivo em adenocarcinoma pancreático. Materiais e métodos: análise clinicopatológica e imuno-histoquímica de 112 pacientes com diagnóstico de câncer de pâncreas com base nos critérios propostos para câncer de mama (HercepTest™) e de estômago (ToGA Trial). Resultados: pelo HercepTest™ 5 (4.5%) casos apresentaram escore 3+, 3 (2.7%) escore 2+ e 104 (92.9%) escores 0/1+. Na análise pelo ToGA Trial, 9 (8.0%) obtiveram escore 3+, 32 (28.6%) escore 2+ e 71 (63.4%) escores 0/1+. Todos os casos positivos pelo HercepTest™ também o foram para o ToGA Trial. Pacientes com hiperexpressão (3+) apresentaram sobrevida média maior que aqueles sem (0 a 2+) tanto pelo HercepTest™ quanto pelo ToGa Trial (43.88 vs. 10.3 meses, p = 0.029 e 40.7 vs. 10.1 meses, p = 0.013, respectivamente). Os demais parâmetros não mostraram correlação com a expressão de HER-2. Conclusão: diferenças na incidência e no significado prognóstico da superexpressão podem ser decorrentes do pequeno tamanho amostral e do uso de dois critérios diferentes de positividade para HER-2. Estes resultados servem com impulso para novas investigações de superexpressão e amplificação do HER-2 utilizando, além da imuno-histoquímica, métodos como FISH e SISH, a fim de se obter mais opções terapêuticas para oferecer aos pacientes, como agentes anti-HER-2. / Introduction: HER-2 overexpression is correlated with aggressiveness in breast and gastric cancers, and its detection has been incorporated as routine in the analysis of these neoplasms. Ideal criteria for evaluation of HER-2 in pancreatic cancer remain unclear. Objectives: to assess the HER-2 status and its predictive value in pancreatic adenocarcinoma. Material and methods: clinicopathologic and immunohistochemical analysis were undertaken in 112 patients with pancreatic cancer using the criteria proposed for breast (HercepTest™) and stomach cancer (ToGA Trial). Results: using HercepTest™ 5 (4.5%) cases had a score of 3+, 3 (2.7%) had a score of 2+ and 104 (92.9%) had scores of 0/1 +. By ToGA Trial, 9 (8.0%) obtained score of 3+, 32 (28.6%) had a score of 2+ score and 71 (63.4%) had scores of 0/1 +. All positive cases by HercepTest™ also went to the ToGA Trial. Patients with overexpression (3 +) showed greater survival than those without (0 to 2 +) by both HercepTest™ and ToGa Trial (43.88 vs. 10.3 months, p = 0.029 and 40.7 vs. 10.1 months, p = 0.013, respectively). Other parameters did not show correlation with the expression of HER-2. Conclusion: differences in incidence and prognostic significance of overexpression may be explained from small sample size and the use of two different criteria of positivity for HER-2. These results serve as an impulse for new investigations of overexpression/amplification of the HER-2 using, besides immunohistochemistry, FISH and SISH methods, in order to get more treatment options to provide patients, as agents anti-HER-2.

abstract: Cancer is a disease that affects millions of people worldwide each year. The metastatic progression of cancer is the number one reason for cancer related deaths. Cancer preventions rely on the early identification of tumor cells as well as a detailed understanding of cancer as a whole. Identifying proteins specific to tumor cells provide an opportunity to develop noninvasive clinical tests and further our understanding of tumor biology. Using liquid chromatography-mass spectrometry (LC-MS/MS) a short peptide was identified in pancreatic cancer patient plasma that was not found in normal samples, and mapped back to QSOX1 protein. Immunohistochemistry was performed probing for QSOX1 in tumor tissue and discovered that QSOX1 is highly over-expressed in pancreatic and breast tumors. QSOX1 is a FAD-dependent sulfhydryl oxidase that is extremely efficient at forming disulfide bonds in nascent proteins. While the enzymology of QSOX1 has been well studied, the tumor biology of QSOX1 has not been studied. To begin to determine the advantage that QSOX1 over-expression provides to tumors, short hairpin RNA (shRNA) were used to reduce the expression of QSOX1 in human tumor cell lines. Following the loss of QSOX1 growth rate, apoptosis, cell cycle and invasive potential were compared between tumor cells transduced with shQSOX1 and control tumor cells. Knock-down of QSOX1 protein suppressed tumor cell growth but had no effect on apoptosis and cell cycle regulation. However, shQSOX1 dramatically inhibited the abilities of both pancreatic and breast tumor cells to invade through Matrigel in a modified Boyden chamber assay. Mechanistically, shQSOX1-transduced tumor cells secreted MMP-2 and -9 that were less active than MMP-2 and -9 from control cells. Taken together, these results suggest that the mechanism of QSOX1-mediated tumor cell invasion is through the post-translational activation of MMPs. This dissertation represents the first in depth study of the role that QSOX1 plays in tumor cell biology. / Dissertation/Thesis / Ph.D. Molecular and Cellular Biology 2012

Introdução: O Adenocarcinoma ductal pancreático é um tumor bastante agressivo que apresenta uma alta de letalidade e, para o qual, poucas opções terapêuticas estão disponíveis. Isto pode ser parcialmente explicado pela complexidade derivada de múltiplas aberrações genéticas e da população celular mista presente em um tumor pancreático, o que também pode explicar o curso clínico heterogêneo observado na prática diária. Ultimamente, pesquisas científicas têm contribuído para ampliar o conhecimento sobre o impacto das alterações epigenéticas no desenvolvimento de múltiplos tipos de câncer, porém no pâncreas essas alterações ainda são incertas e, por isso, foco de investigação. A desregulação epigenética parece estar envolvida no ciclo celular da célula tumoral, incluindo o crescimento celular, diferenciação, progressão tumoral e morte celular, e a acetilação das histonas é um importante mecanismo que regula a transcrição de genes envolvidos nesses processos. Padrões globais de modificações das histonas foram recentemente apontados como preditores de desfecho em pacientes com câncer, mas poucos estudos têm sido realizados nesta área, inclusive em Adenocarcinoma ductal pancreático (ADP). Objetivos: O presente estudo foi desenvolvido a fim de investigar o padrão de modificação de acetilação das histonas em adenocarcinoma pancreático, através da análise imunohistoquímica. Materiais e métodos: Uma análise clinicopatológica retrospectiva foi realizada em 119 pacientes diagnosticados com câncer de pâncreas entre os anos de 2005 e 2011, e realizado estudo imunohistoquímico com os anticorpos contra H4K12ac, H3K9ac e H3K18ac. Marcação nuclear positiva para cada histona foi medida quanto à intensidade e expressão, sendo classificadas em grupos de baixa ou de alta intensidade/expressão. Os resultados foram analisados em relação aos parâmetros clinicopatológicos de cada paciente. Resultados: Houve uma relação positiva entre diferenciação tumoral e alta expressão de H4K12ac (P <0,05), bem como a intensidade forte dos três marcadores correlacionou-se positivamente com o estágio do tumor (P <0,01). Análise univariada mostrou pior sobrevida em pacientes com níveis elevados de expressão de H4K12ac (p = 0,038) e H3K18Ac (P = 0,033). Modelo de risco proporcional de Cox revelou o efeito prognóstico independente de níveis elevados de H4K12ac H3K18ac (taxas de risco de 1,6 e 1,7, respectivamente, p <0,05), especialmente para pacientes em estágios iniciais. Sugerimos como hipótese que as modificações na acetilação das histonas H4K12 e H3K18 podem ser consideradas fatores prognósticos importantes para o câncer de pâncreas, embora o mecanismo envolvido necessite de mais investigação. Aumentando a compreensão e o conhecimento sobre o padrão de acetilação das histonas, poder-se-ão finalmente gerar novas idéias para um diagnóstico molecular racional e novas abordagens terapêuticas. / Introduction: Ductal pancreatic adenocarcinoma (DPAs) is a highly aggressive tumor, with a high letality rate, for which few therapeutic options are available. This may be partially explained by the notorious complexity derived from the multiple genetic aberrations and mixed cellular population present in a pancreatic tumor, which can also explain the heterogeneous clinical course observed in daily practice. Lately, there is an increase in the literature about the impact of epigenetic changes on the development of several cancer, however in the pancreas these changes are still uncertain. Epigenetic deregulation may be involved in tumor cell biology, including cell growth, differentiation, tumor progression and cell death, and histone acetylation is a major mechanism that regulates gene transcription. Patterns of global histone modifications have been recently suggested as outcome predictors in cancer patients, but few studies have been conducted on pancreatic ductal adenocarcinomas. Objectives: This study was designed to investigate the predictive value of histone acetylation modifications on pancreatic cancer. Material and methods: A retrospective clinicopathologic analysis was undertaken in 119 patients diagnosed with PDAC between 2005 and 2011, and immunohistochemistry performed with antibodies against H4K12ac, H3K9ac and H3K18ac. Positive nuclear staining for each histone was measured as the intensity and expression, being classified into low or high-staining groups. Results were analyzed in relation to patients’ clinicopathologic parameters. Results: There was a positive relationship between tumor differentiation and H4K12ac high scores (P<0.05) and staining of the three markers correlated positively with tumor stage (P<0.01). Univariate analysis showed worse survival in patients with high detection levels of H4K12ac (p=0.038) and H3K18Ac (P=0.033). A backwards Cox proportional hazards model revealed the independent prognostic effect of high H4K12ac and H3K18ac levels (hazard ratios of 1.6 and 1.7 respectively, p<0.05), especially for patients at early stages. We hypothesize that acetylation of H4K12 and H3K18 may be considered valuable prognostic factors for pancreatic cancer, although the mechanism involved needs further investigation. Increasing insights into histone acetylation modifications can ultimately generate new ideas for rationally and molecularly based diagnostic and therapeutic approaches.

Introdução: a superexpressão de HER-2 tem correlação com maior agressividade em carcinomas de mama e estômago, e a sua detecção já foi incorporada como rotina na análise destas neoplasias. Critérios ideais para avaliação do HER-2 em câncer de pâncreas permanecem incertos. Objetivos: avaliar o status do HER-2 e o seu valor preditivo em adenocarcinoma pancreático. Materiais e métodos: análise clinicopatológica e imuno-histoquímica de 112 pacientes com diagnóstico de câncer de pâncreas com base nos critérios propostos para câncer de mama (HercepTest™) e de estômago (ToGA Trial). Resultados: pelo HercepTest™ 5 (4.5%) casos apresentaram escore 3+, 3 (2.7%) escore 2+ e 104 (92.9%) escores 0/1+. Na análise pelo ToGA Trial, 9 (8.0%) obtiveram escore 3+, 32 (28.6%) escore 2+ e 71 (63.4%) escores 0/1+. Todos os casos positivos pelo HercepTest™ também o foram para o ToGA Trial. Pacientes com hiperexpressão (3+) apresentaram sobrevida média maior que aqueles sem (0 a 2+) tanto pelo HercepTest™ quanto pelo ToGa Trial (43.88 vs. 10.3 meses, p = 0.029 e 40.7 vs. 10.1 meses, p = 0.013, respectivamente). Os demais parâmetros não mostraram correlação com a expressão de HER-2. Conclusão: diferenças na incidência e no significado prognóstico da superexpressão podem ser decorrentes do pequeno tamanho amostral e do uso de dois critérios diferentes de positividade para HER-2. Estes resultados servem com impulso para novas investigações de superexpressão e amplificação do HER-2 utilizando, além da imuno-histoquímica, métodos como FISH e SISH, a fim de se obter mais opções terapêuticas para oferecer aos pacientes, como agentes anti-HER-2. / Introduction: HER-2 overexpression is correlated with aggressiveness in breast and gastric cancers, and its detection has been incorporated as routine in the analysis of these neoplasms. Ideal criteria for evaluation of HER-2 in pancreatic cancer remain unclear. Objectives: to assess the HER-2 status and its predictive value in pancreatic adenocarcinoma. Material and methods: clinicopathologic and immunohistochemical analysis were undertaken in 112 patients with pancreatic cancer using the criteria proposed for breast (HercepTest™) and stomach cancer (ToGA Trial). Results: using HercepTest™ 5 (4.5%) cases had a score of 3+, 3 (2.7%) had a score of 2+ and 104 (92.9%) had scores of 0/1 +. By ToGA Trial, 9 (8.0%) obtained score of 3+, 32 (28.6%) had a score of 2+ score and 71 (63.4%) had scores of 0/1 +. All positive cases by HercepTest™ also went to the ToGA Trial. Patients with overexpression (3 +) showed greater survival than those without (0 to 2 +) by both HercepTest™ and ToGa Trial (43.88 vs. 10.3 months, p = 0.029 and 40.7 vs. 10.1 months, p = 0.013, respectively). Other parameters did not show correlation with the expression of HER-2. Conclusion: differences in incidence and prognostic significance of overexpression may be explained from small sample size and the use of two different criteria of positivity for HER-2. These results serve as an impulse for new investigations of overexpression/amplification of the HER-2 using, besides immunohistochemistry, FISH and SISH methods, in order to get more treatment options to provide patients, as agents anti-HER-2.

Le cancer du pancréas est un cancer très agressif et de pronostic très sombre. Il est diagnostiqué tardivement et se montre résistant aux traitements. Dans ce contexte, il est nécessaire de mettre en évidence de nouveaux marqueurs spécifiques de cette pathologie dévastatrice. Le but de notre étude était de rechercher un marqueur « génétique » de ce cancer au sein du gène de la lipase sels biliaires-dépendante (BSDL) localisé en position 9q34.3 et plus particulièrement au niveau de l'exon 11 de ce gène codant pour le domaine C-terminal de la protéine et constitué de séquences répétées (séquences VNTR). L’analyse des électrophérogrammes obtenues après séquençage de Sanger à partir des amplicons de PCR réalisées sur l’ADNg extraits de tissus de patients atteints de cancer du pancréas, a permis d’identifier deux altérations génétiques : (i) la présence d’un SNP (Single Nucleotide Polymorphism) synonyme référencé rs488087, impliquant la transition c.1719C>T ; qui semble être associée au développement d’un cancer du pancréas sporadique et pourrait être un potentiel facteur prédictif de ce cancer permettant de cibler des populations à risque, (ii) la présence d’une insertion d’un nucléotide C induisant l'apparition d'une protéine BSDL tronquée présentant une séquence C-terminale modifiée contre laquelle ont été développé des anticorps polyclonaux. Cette nouvelle séquence pourrait constituer un potentiel marqueur diagnostique et/ou thérapeutique du cancer du pancréas. Ces deux altérations génétiques constituent ainsi de potentiels marqueurs du cancer du pancréas. / Pancreatic cancer is a devastating disease progressing asymptomatically until death within months after diagnosis. In this context, it is necessary to identify new specific markers to develop diagnostic tools and to target an at risk population. The aim of our study was to find a "genetic" marker in the bile salt-dependant lipase gene (BSDL). The human BSDL gene is located on the long arm of chromosome 9 in 9q34.3 with a variable number of tandem repeats (VNTR) in the coding region of exon 11. The electropherograms obtained after Sanger sequencing analysis of gDNA amplified from pancreatic cancer tissue samples allowed us to highlight: (i) the presence of a SNP (Single Nucleotide Polymorphism) involved in c.1719C>T transition which is referenced rs488087. rs488087 seems to be associated with the sporadic pancreatic cancer development and may be a predictive factor of pancreatic cancer for targeting an at risk population, (ii) the presence of a C nucleotide insertion leading to a premature stop codon with truncated protein and to the modification of the C-terminal sequence end. This new C-terminal sequence, alteration could be used as a potential diagnostic and/or therapeutic marker. Finally, these two genetic alterations identified in BSDL gene could constitute potential markers of pancreatic cancer.

Pancreatic cancer is the fourth leading cause of cancer death in Canada. Significant advancements in chemotherapy for advanced pancreatic cancer have resulted in the need for a quantitative comparison between these treatments on a relative scale. Therefore, a systematic review and Bayesian network meta-analysis of randomized clinical trials was conducted using gemcitabine, the standard treatment, as the reference comparator. Based on results from the network meta-analysis, in which optimal treatments were identified and side effects of each treatment evaluated, an Internet-based patient decision aid was developed in order to present the benefits and risks of each therapy option: (1) Best supportive care (2) gemcitabine (3) FOLFIRINOX. The objective of the decision aid was to guide patients through the decision-making process based on their individual preferences and values. The decision aid was deemed to be acceptable and feasible based on results from a pilot study conducted at The Ottawa Hospital Cancer Centre.

With a five-year survival rate of less than 5%, pancreatic cancer is one of the deadliest cancers. The most common activating mutations in pancreatic cancer are found in the KRAS gene, causing a constitutively-active KRAS protein in approximately 90% of pancreatic ductal adenocarcinomas (PDAC). PDAC-derived cell lines that harbor oncogenic KRAS mutations can be divided into two classes, KRAS dependent (or addicted) cells and KRAS independent cells. Oncogene dependency (or addiction) is a phenomenon where tu-mors require sustained activity of a single aberrantly activated gene despite the accumulation of multiple oncogenic lesions. In the case of PDAC, the single aberrantly activated gene is KRAS. KRAS independent cells have acquired various other oncogenic lesions that confer alternative cell survival signaling pathways to bypass oncogenic KRAS dependency. The Spleen Tyrosine Kinase (Syk) is highly expressed in KRAS dependent cells, while KRAS independent cells have low Syk expression. This pattern suggests that in KRAS dependent cells, constitutively active KRAS and Syk play a role in stimulating pro-survival pathways. One of these pro-survival pathways is known as mTORC1, which causes increased anabolic processes like protein and lipid synthesis. Accordingly, mTORC1 causes suppression of catabolic processes like autophagy. The net effect is in-creased cellular growth and proliferation. However, mTORC1 inhibitors have limited clinical efficacy, and potential therapeutic targets upstream of mTORC1 have drawn interest. Syk is a non-receptor tyrosine kinase that is an upstream activator of the mTORC1 pathway in hematopoietic malignancies. Through Syk inhibition studies using the small molecule PRT062607 (SYKi), we demonstrated that Syk is also involved in activating the mTORC1 pathway in KRAS dependent PDAC cells. However, the mechanism by which Syk-mediated activation of mTORC1 occurs is currently unknown. Moreover, it is unclear whether SYK kinase activity is required for the activation of the mTORC1 pathway. To address this issue, we introduced a single nucleotide mutation in the kinase do-main of Syk to render it kinase-inactive and found that Syk requires its kinase function to activate mTORC1. Studies using Syki also revealed that mTORC1 activity was also inhibited in KRAS independent PDAC cells that lack significant Syk expression. Interestingly, substrate specificity studies indicate that Syki also binds to and inhibits structurally similar protein tyrosine kinases such as the SRC Family Kinases (SFKs). Therefore, we designed an experiment to look for Syk and SFK cooperativity in regards to mTORC1 activation in PDAC cells. Our results indicate that the SFKs, Yes1 and Src display the most significant cooperative effect with Syk in activating the mTORC1 pathway. Src and Yes1 may even be involved in the upstream activation of Syk. To establish the physiological significance of Syk signaling in pancreatic cancer, it is important to establish model organisms that could be used for future studies. Thus, we test-ed Syk expression and function in PDAC cell lines derived from genetically-engineered mouse models (GEMM), which develop pancreatic cancer via oncogenic mutations in KRAS and TP53. We found that Syk is indeed expressed in murine PDAC cell lines and that the use of Syki in the murine PDAC cell lines results in decreased mTORC1 activity. These results recapitulate those obtained in human KRAS dependent PDAC cell lines. In summary, our studies show that Syk is a key regulator of mTORC1 signaling in human and mouse-derived pancreatic cancer cells. Syk kinase activity is required for mTORC1 activation. Finally, SFKs cooperate with Syk to promote robust mTORC1 activation. The mechanisms of SFK and Syk cooperativity in mTORC1 pathway activation will require further investigation. Additionally, our findings provide a strong rationale to study the effects of Syk kinase inhibition in physiologically-relevant murine models of pancreatic cancer. / 2021-06-08T00:00:00Z