Orientador: Lígia Yukie sassaki / Resumo: Introdução: Com o advento da terapia biológica, o foco de resposta na Doença de Crohn (DC) ampliou-se, sendo essenciais a remissão clínica e endoscópica. Estudos prospectivos comparando os representantes dessa classe terapêutica mais utilizados, o Infliximabe (IFX) e o Adalimumabe (ADA), são escassos. O objetivo do estudo foi comparar a efetividade clínica e endoscópica do IFX versus ADA em pacientes com DC naïves terapia biológica, na semana 54 de tratamento. Metodologia: Foi realizado um estudo clínico, não-randomizado, no qual pacientes com DC que receberam IFX ou ADA, foram avaliados nas semanas 0, 14, 30 e 54 de tratamento. Utilizou-se o Índice de Atividade da Doença de Crohn (CDAI) para avaliação da atividade clínica da doença. Entre 6 e 12 meses, foi avaliada a atividade endoscópica através do Simplified Endoscopic Score for Crohn’s Disease (SES-CD). As variáveis foram resposta clínica (queda do CDAI > 70 pontos) e remissão clínica (CDAI < 150 pontos), avaliados nas semanas 14, 30 e 54, e resposta endoscópica (queda do SES-CD de pelo menos 50% em relação à pontuação inicial), remissão endoscópica (controle endoscópico com SES-CD ≤ 2 pontos), e taxas de internação, cirurgia, óbito e perda de resposta, analisados na semana 54. A análise estatística foi estatística descritiva, teste de ANOVA com medidas repetidas no tempo considerando a interação medicamento x tempo, seguida do teste de comparação múltipla de Tukey ajustado, com nível de significância de 5% ou o p-valor c... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: With the advent of biological therapy, the focus of the Crohn's Disease (CD) response has changed and clinical and endoscopic remission are essential. Prospective studies comparing the most used biological drugs, Infliximab (IFX) and Adalimumab (ADA), are scarce. Our aim was compare the clinical and endoscopic effectiveness of IFX versus ADA in CD patients naïve to biological therapy at the 54th week of treatment. Methods: An open, non-randomized, clinical study with CD patients receiving IFX or ADA was performed at treatment weeks 0, 14, 30 and 54. The Crohn's Disease Activity Index (CDAI) was used to evaluate the disease clinical activity. Between 6 and 12 months, colonoscopy was performed and the Simplified Endoscopic Score for Crohn's Disease (SES-CD) was used. Clinical response (CDAI decrease>70 points) and clinical remission (CDAI<150 points) were assessed at weeks 14, 30 and 54; endoscopic response (decrease of at least 50% in SES-CD), endoscopic remission (endoscopic control with SES-CD ≤ 2 points), and hospitalization rates, surgeries, deaths and loss of response, were analyzed at week 54. A statistical analysis was descriptive with ANOVA test with repeated measures considering drug interaction x time, followed by Tukey multiple comparison test adjusted, with a significance level of 5% or the corresponding p-value. Results: A total of 85 patients were included, 45 patients underwent treatment with ADA and 40 with IFX, with difference between groups only... (Complete abstract click electronic access below) / Doutor

Submitted by JÚLIO PINHEIRO BAIMA (jpbaima@yahoo.com.br) on 2018-04-25T20:09:46Z No. of bitstreams: 1 Tese Doutorado Julio Pinheiro Baima 25 abril 2018.pdf: 1976761 bytes, checksum: fade6f9adcb14ae4ad0854e9274f5c9d (MD5) / Approved for entry into archive by Sulamita Selma C Colnago null (sulamita@btu.unesp.br) on 2018-04-26T19:04:00Z (GMT) No. of bitstreams: 1 baima_jp_dr_bot_int.pdf: 1976761 bytes, checksum: fade6f9adcb14ae4ad0854e9274f5c9d (MD5) / Made available in DSpace on 2018-04-26T19:04:00Z (GMT). No. of bitstreams: 1 baima_jp_dr_bot_int.pdf: 1976761 bytes, checksum: fade6f9adcb14ae4ad0854e9274f5c9d (MD5) Previous issue date: 2018-02-27 / Introdução: Com o advento da terapia biológica, o foco de resposta na Doença de Crohn (DC) ampliou-se, sendo essenciais a remissão clínica e endoscópica. Estudos prospectivos comparando os representantes dessa classe terapêutica mais utilizados, o Infliximabe (IFX) e o Adalimumabe (ADA), são escassos. O objetivo do estudo foi comparar a efetividade clínica e endoscópica do IFX versus ADA em pacientes com DC naïves terapia biológica, na semana 54 de tratamento. Metodologia: Foi realizado um estudo clínico, não-randomizado, no qual pacientes com DC que receberam IFX ou ADA, foram avaliados nas semanas 0, 14, 30 e 54 de tratamento. Utilizou-se o Índice de Atividade da Doença de Crohn (CDAI) para avaliação da atividade clínica da doença. Entre 6 e 12 meses, foi avaliada a atividade endoscópica através do Simplified Endoscopic Score for Crohn’s Disease (SES-CD). As variáveis foram resposta clínica (queda do CDAI > 70 pontos) e remissão clínica (CDAI < 150 pontos), avaliados nas semanas 14, 30 e 54, e resposta endoscópica (queda do SES-CD de pelo menos 50% em relação à pontuação inicial), remissão endoscópica (controle endoscópico com SES-CD ≤ 2 pontos), e taxas de internação, cirurgia, óbito e perda de resposta, analisados na semana 54. A análise estatística foi estatística descritiva, teste de ANOVA com medidas repetidas no tempo considerando a interação medicamento x tempo, seguida do teste de comparação múltipla de Tukey ajustado, com nível de significância de 5% ou o p-valor correspondente. Resultados: Foram incluídos 85 pacientes, 45 submetidos ao tratamento com ADA e 40 com IFX. O uso concomitante de Azatioprina foi mais frequente no grupo que recebeu IFX (p=0,0001). As taxas de resposta clínica foram de 86,67% na semana 14, 82,22% na semana 30 e 82,22% na semana 54 no grupo ADA. No grupo IFX, as taxas de resposta foram 70%, 72,5% e 75%, respectivamente, sem diferença entre os tratamentos (p>0,05 em todas as semanas). As taxas de remissão clínica no grupo ADA foram de 86,67% na semana 14, 80% na semana 30 e 82,22% na Resumo 3 semana 54. No grupo IFX, 65%, 62,5% e 65% respectivamente, com diferença significativa apenas na semana 14 (p=0,02). Resposta endoscópica foi atingida em 60,61% dos pacientes do grupo ADA e 85,71% no grupo IFX (p=0,02). As taxas de remissão endoscópica foram de 45,45% e 60%, respectivamente (p=0,23). Internações, cirurgia, óbitos e perda de resposta ocorreram com frequência sem diferença significativa entre os grupos. Conclusões: Não houve diferença nas taxas de resposta clínica entre os tratamentos. Houve maiores taxas de remissão clínica na semana 14 no grupo ADA, não mantidas nas semanas seguintes. Taxas de resposta endoscópica foram melhores no grupo IFX. Não houve diferença nas taxas de remissão endoscópica. Não houve diferença nas taxas de internações, cirurgias e óbitos, assim como na perda de resposta. / Introduction: With the advent of biological therapy, the focus of the Crohn's Disease (CD) response has changed and clinical and endoscopic remission are essential. Prospective studies comparing the most used biological drugs, Infliximab (IFX) and Adalimumab (ADA), are scarce. Our aim was compare the clinical and endoscopic effectiveness of IFX versus ADA in CD patients naïve to biological therapy at the 54th week of treatment. Methods: An open, non-randomized, clinical study with CD patients receiving IFX or ADA was performed at treatment weeks 0, 14, 30 and 54. The Crohn's Disease Activity Index (CDAI) was used to evaluate the disease clinical activity. Between 6 and 12 months, colonoscopy was performed and the Simplified Endoscopic Score for Crohn's Disease (SES-CD) was used. Clinical response (CDAI decrease>70 points) and clinical remission (CDAI<150 points) were assessed at weeks 14, 30 and 54; endoscopic response (decrease of at least 50% in SES-CD), endoscopic remission (endoscopic control with SES-CD ≤ 2 points), and hospitalization rates, surgeries, deaths and loss of response, were analyzed at week 54. A statistical analysis was descriptive with ANOVA test with repeated measures considering drug interaction x time, followed by Tukey multiple comparison test adjusted, with a significance level of 5% or the corresponding p-value. Results: A total of 85 patients were included, 45 patients underwent treatment with ADA and 40 with IFX, with difference between groups only in concomitant use of azathioprine, more frequent in the IFX group (p=0.0001). Clinical response rates were 86.67% at week 14, 82.22% at week 30 and 82.22% at week 54 in ADA group. In IFX group, 70%, 72.5% and 75%, respectively (p> 0.05 on all weeks). Clinical remission rates were 86.67% at week 14, 80% at week 30 and 82.22% at week 54 in ADA group. In IFX group, 65%, 62.5% and 65%, respectively, with significate difference only at week 14 (p = 0.02). Endoscopic response rate was 60.61% in patients from ADA group and 85.71% in the IFX group (p = 0.02). Rates of Abstract 6 endoscopic remission were 45.45% and 60%, respectively (p = 0.23). Hospitalizations, surgery, deaths and loss of lost response occurred without significant difference between groups. Conclusions: There were no difference in the clinical response rates between treatments. There was higher rate of clinical remission at week 14 in the ADA group, not sustained in the subsequent weeks. Endoscopic response rate was better in the IFX group. There was no difference in endoscopic remission rate. There were no difference in hospitalization rates, surgeries and deaths, as well as loss of response.

A infecção de cães pela Leishmania (Leishmania) infantum resulta em um espectro de manifestações imunopatológicas que dependem da interação parasito hospedeiro e são definidas por fatores ambientais e pela genética do hospedeiro. Apesar disso, a imunogenética da leishmaniose visceral canina (LVC) permanece inexplorada. Nós realizamos diagnóstico laboratorial, clínico, ensaio de linfoproliferação (LPA), quantificação de citocinas, teste de hipersensibilidade tardia à leishmanina (LST), quantificação de IgA, IgE, IgG, IgM anti L. (L. ) infantum, IgG anti saliva de flebotomíneo e genotipagem ampla afim de identificar polimorfismos de nucleotídeo único (SNPs) associados aos diferentes perfis de imunidades celular, humoral, resposta clínica e status de infecção em cães de área endêmica, utilizando modelo de componente de variância (EMMAX). O efeito de estrutura da amostra foi controlado em todas as análises. A presença ou ausência de infecção pela L. (L. ) infantum foi associado a regiões contendo os genes PRGR_CANFA, RAB38, NOX4, PRKCI e SMAD7, IL1RA, IL12A_CANFA relacionados à ativação de fagócitos, mecanismos microbicidas, sobrevivência intracelular de patógenos e resposta pró inflamatória; a resposta clínica foi associada a regiões contendo os genes CATA_CANFA, LIAS, IL17A e IL17F relacionados à proteção contra danos do estresse oxidativo e indução de resposta pró inflamatória; o resultado LST+ foi associado à resposta Th1, controle da infecção mas não preveniu a manifestação de sinais clínicos, enquanto o LST- foi associado à resposta Treg e aumento do parasitismo. O resultado do LST foi associado a regiões contendo os genes MEP1B, PTPRM, TLN1, TGFBR1, ITGA9, EPCAM e CALM1 envolvidos com maturação de fagócitos, migração e adesão de leucócitos, estabilidade da sinapse imunológica, diferenciação e proliferação de linfócitos. A linfoproliferação foi dependente da carga parasitária e associada a regiões contendo os genes FOCAD, PIAS2, SMAD2 e IL6R envolvidos com supressão tumoral e diferenciação de linfócitos Th17 ou Treg; o aumento dos níveis de IgA, IgE e IgG anti L. (L. ) infantum foi associado à LVC sintomática enquanto o de IgG anti saliva de Lutzomyia longipalpis foi associado à exposição e infecção assintomática. Quanto à resposta de IgM, foram identificados SNPs nos genes NXN e SH3BP5 relacionados com inibição do crescimento, diferenciação e ativação de linfócitos B e vias de sinalização de TLR4 e TLR9; para IgG anti - L. (L. ) infantum foram identificadas regiões contendo os genes IL17RB, SH2B3 e replicação do loci de susceptibilidade NOX4, RAB38, CTSC envolvidos com linfopoiese, citocinese, resposta pró inflamatória, mecanismos microbicidas, sobrevivência intracelular de Leishmania; os níveis de IgA foram associados a regiões contendo os genes LIN28A e MAFB implicados na predisposição à nefropatia glomerular, já os níveis de IgG anti saliva de Lu. longipalpis foram associados a regiões contendo os genes ERBB2IP, CD180, RAB7A_CANFA, FOXP1, RUNX1, SOD1, Q3HTU8_CANFA, IFNAR1, IFNAR2 e IFNGR2 envolvidos com supressão celular, produção de imunoglobulina via TLR4 e sobrevivência intracelular de Leishmania. Esses resultados apontam regiões cromossômicas úteis para a elucidação da resposta à infecção por L. (L. ) infantum em cães e alvos potenciais para estudos funcionais, estratégias profiláticas e terapêuticas / Leishmania (Leishmania) infantum infection in dogs leads to a range of immunopathological responses, which depend on a parasite - host interaction and are defined by environmental factors and genetic of host. Neverthless, immunogenetic of the canine leishmaniasis (CanL) remains unexplored. We performed diagnosis and clinical evaluation, lymphoproliferation assay (LPA), leishmanin skin test (LST), quantification of cytokine, anti-L. (L. ) infantum IgA, IgE, IgG, IgM, anti- sandfly saliva IgG levels and genome wide association scan of 110.165 SNPs (GWAS) in order to indentify loci associated to clinical, antibody, cell-mediated responses and status of infection in 189 dogs, employng a expedited efficient mix model of association (EMMAX). Control of stratification effects due to sample structure was evideced by the low inflation factors. Status of infection was associated to SNPs in linkage desequilibrium (LD) with PRGR_CANFA, RAB38, NOX4, PRKCI and in the neighborhood of SMAD7, IL1RA, IL12A_CANFA genes related to phagocyte maturation, killing of pathogens and proinflamatory response; clinical outcome was associated to CATA_CANFA, LIAS, IL17A, IL17F loci involved in prevention of oxidative burst mediated injury and proinflamatory response; LST+ was associated to Th1 response although it has not prevented symptoms, whereas LST- was associated to Treg response and enhanced parasite load. Overall, LST response was associated to MEP1B, PTPRM, TLN1, TGFBR1, ITGA9, EPCAM e CALM1 loci committed to phagocyte maturation, leukocyte adhesion and migration, stability in immunological synapse, lymphocyte diferenciation and proliferation. Lymphocyte proliferation was rely on parasit burden and associated to FOCAD, PIAS2 loci in the neighborhood of SMAD2 e IL6R genes, wich are implicated in tumor supression and Treg/Th17 decision; Increased levels of anti-L. (L. ) infantum IgA, IgE, IgG were observed in severity of CanL. In contrast, anti- sandfly saliva IgG was enhenced in asymptomatic dogs. IgM response was associated to NXN e SH3BP5 loci related to fate, growing and activation of B cells; anti-L. (L. ) infantum IgG levels was associated to region containing IL17RB, SH2B3 and replication of NOX4, RAB38, CTSC suscptibility loci involved in proinflamatory response, microbicidal activity, lymphopoiesis and cytokinesis; IgA levels were associated to SNPs on LIN28A and MAFB, wich are implicated glomerular nephropathy, whereas anti-sandfly saliva IgG levels were associated to ERBB2IP, CD180, RAB7A_CANFA, FOXP1, RUNX1, SOD1, Q3HTU8_CANFA, IFNAR1, IFNAR2 e IFNGR2 loci, wich are related to supression of inflamation, antibody response through the TLR4 pathway and survival of intracellular pathogens. These findins provide insights for responses to L. (L. ) infantum infection and point to potential targets for functional investigations, therapeutic and prophylactic strategies

A infecção de cães pela Leishmania (Leishmania) infantum resulta em um espectro de manifestações imunopatológicas que dependem da interação parasito hospedeiro e são definidas por fatores ambientais e pela genética do hospedeiro. Apesar disso, a imunogenética da leishmaniose visceral canina (LVC) permanece inexplorada. Nós realizamos diagnóstico laboratorial, clínico, ensaio de linfoproliferação (LPA), quantificação de citocinas, teste de hipersensibilidade tardia à leishmanina (LST), quantificação de IgA, IgE, IgG, IgM anti L. (L. ) infantum, IgG anti saliva de flebotomíneo e genotipagem ampla afim de identificar polimorfismos de nucleotídeo único (SNPs) associados aos diferentes perfis de imunidades celular, humoral, resposta clínica e status de infecção em cães de área endêmica, utilizando modelo de componente de variância (EMMAX). O efeito de estrutura da amostra foi controlado em todas as análises. A presença ou ausência de infecção pela L. (L. ) infantum foi associado a regiões contendo os genes PRGR_CANFA, RAB38, NOX4, PRKCI e SMAD7, IL1RA, IL12A_CANFA relacionados à ativação de fagócitos, mecanismos microbicidas, sobrevivência intracelular de patógenos e resposta pró inflamatória; a resposta clínica foi associada a regiões contendo os genes CATA_CANFA, LIAS, IL17A e IL17F relacionados à proteção contra danos do estresse oxidativo e indução de resposta pró inflamatória; o resultado LST+ foi associado à resposta Th1, controle da infecção mas não preveniu a manifestação de sinais clínicos, enquanto o LST- foi associado à resposta Treg e aumento do parasitismo. O resultado do LST foi associado a regiões contendo os genes MEP1B, PTPRM, TLN1, TGFBR1, ITGA9, EPCAM e CALM1 envolvidos com maturação de fagócitos, migração e adesão de leucócitos, estabilidade da sinapse imunológica, diferenciação e proliferação de linfócitos. A linfoproliferação foi dependente da carga parasitária e associada a regiões contendo os genes FOCAD, PIAS2, SMAD2 e IL6R envolvidos com supressão tumoral e diferenciação de linfócitos Th17 ou Treg; o aumento dos níveis de IgA, IgE e IgG anti L. (L. ) infantum foi associado à LVC sintomática enquanto o de IgG anti saliva de Lutzomyia longipalpis foi associado à exposição e infecção assintomática. Quanto à resposta de IgM, foram identificados SNPs nos genes NXN e SH3BP5 relacionados com inibição do crescimento, diferenciação e ativação de linfócitos B e vias de sinalização de TLR4 e TLR9; para IgG anti - L. (L. ) infantum foram identificadas regiões contendo os genes IL17RB, SH2B3 e replicação do loci de susceptibilidade NOX4, RAB38, CTSC envolvidos com linfopoiese, citocinese, resposta pró inflamatória, mecanismos microbicidas, sobrevivência intracelular de Leishmania; os níveis de IgA foram associados a regiões contendo os genes LIN28A e MAFB implicados na predisposição à nefropatia glomerular, já os níveis de IgG anti saliva de Lu. longipalpis foram associados a regiões contendo os genes ERBB2IP, CD180, RAB7A_CANFA, FOXP1, RUNX1, SOD1, Q3HTU8_CANFA, IFNAR1, IFNAR2 e IFNGR2 envolvidos com supressão celular, produção de imunoglobulina via TLR4 e sobrevivência intracelular de Leishmania. Esses resultados apontam regiões cromossômicas úteis para a elucidação da resposta à infecção por L. (L. ) infantum em cães e alvos potenciais para estudos funcionais, estratégias profiláticas e terapêuticas / Leishmania (Leishmania) infantum infection in dogs leads to a range of immunopathological responses, which depend on a parasite - host interaction and are defined by environmental factors and genetic of host. Neverthless, immunogenetic of the canine leishmaniasis (CanL) remains unexplored. We performed diagnosis and clinical evaluation, lymphoproliferation assay (LPA), leishmanin skin test (LST), quantification of cytokine, anti-L. (L. ) infantum IgA, IgE, IgG, IgM, anti- sandfly saliva IgG levels and genome wide association scan of 110.165 SNPs (GWAS) in order to indentify loci associated to clinical, antibody, cell-mediated responses and status of infection in 189 dogs, employng a expedited efficient mix model of association (EMMAX). Control of stratification effects due to sample structure was evideced by the low inflation factors. Status of infection was associated to SNPs in linkage desequilibrium (LD) with PRGR_CANFA, RAB38, NOX4, PRKCI and in the neighborhood of SMAD7, IL1RA, IL12A_CANFA genes related to phagocyte maturation, killing of pathogens and proinflamatory response; clinical outcome was associated to CATA_CANFA, LIAS, IL17A, IL17F loci involved in prevention of oxidative burst mediated injury and proinflamatory response; LST+ was associated to Th1 response although it has not prevented symptoms, whereas LST- was associated to Treg response and enhanced parasite load. Overall, LST response was associated to MEP1B, PTPRM, TLN1, TGFBR1, ITGA9, EPCAM e CALM1 loci committed to phagocyte maturation, leukocyte adhesion and migration, stability in immunological synapse, lymphocyte diferenciation and proliferation. Lymphocyte proliferation was rely on parasit burden and associated to FOCAD, PIAS2 loci in the neighborhood of SMAD2 e IL6R genes, wich are implicated in tumor supression and Treg/Th17 decision; Increased levels of anti-L. (L. ) infantum IgA, IgE, IgG were observed in severity of CanL. In contrast, anti- sandfly saliva IgG was enhenced in asymptomatic dogs. IgM response was associated to NXN e SH3BP5 loci related to fate, growing and activation of B cells; anti-L. (L. ) infantum IgG levels was associated to region containing IL17RB, SH2B3 and replication of NOX4, RAB38, CTSC suscptibility loci involved in proinflamatory response, microbicidal activity, lymphopoiesis and cytokinesis; IgA levels were associated to SNPs on LIN28A and MAFB, wich are implicated glomerular nephropathy, whereas anti-sandfly saliva IgG levels were associated to ERBB2IP, CD180, RAB7A_CANFA, FOXP1, RUNX1, SOD1, Q3HTU8_CANFA, IFNAR1, IFNAR2 e IFNGR2 loci, wich are related to supression of inflamation, antibody response through the TLR4 pathway and survival of intracellular pathogens. These findins provide insights for responses to L. (L. ) infantum infection and point to potential targets for functional investigations, therapeutic and prophylactic strategies

Un défi majeur en oncologie clinique est de caractériser avec précision la réponse des patients aux agents thérapeutiques. Actuellement, il n'existe pas de modèles et de tests fiables capable de reproduire précisément une tumeur primaire dans toute sa complexité. Or, ce paramètre est essentiel pour mettre en œuvre une stratégie de médecine personnalisée capable d'identifier le régime de traitement le plus approprié pour un patient particulier dans un délai cliniquement pertinent. Pour répondre à ce besoin, notre groupe a développé un nouveau modèle 3D ex vivo qui repose sur la micro-dissection d'un échantillon de tumeur (MDT) d'un patient et l'utilisation de technologies microfluidiques pour maintenir la viabilité du tissu et le microenvironnement tumoral naturel afin d’évaluer la sensibilité aux traitements dans un délai adapté à la prise de décision clinique. Cette approche permettrait de sélectionner les thérapies les plus efficaces tout en réduisant l'administration de traitements inefficaces associés à des effets secondaires indésirables, ainsi que les coûts de prise en charge des patients. Des travaux précédemment publiés par notre équipe ont montré que la viabilité des cellules cancéreuses situées dans notre modèle de tumeur ex vivo pouvait être caractérisée par microscopie confocale sur l’intégralité du MDT ou par cytométrie de flux sur les MDTs après dissociation enzymatique des cellules. Cependant, ces techniques présentent des limitations en termes de résolution visuelle pour la microscopie confocale et de sensibilité et information spatiale pour la cytométrie de flux. Nous proposons ici d’associer notre modèle 3D de MDTs en microfluidiques à des techniques d’immuno-histopathologie, dans le but d’offrir une évaluation moléculaire, spatiale et quantitative de la réponse de la tumeur au traitement. Pour cela, nous avons optimisé une procédure de lithographie en paraffine de nos systèmes microfluidiques, permettant la production de blocs de micro-étalages micro-réseaux de tissus micro-disséqués (MDTMA). afin de permettre une coloration morphologique du tissu et un marquage de protéines spécifiques pour analyser l'architecture tissulaire, la prolifération et l’apoptose cellulaire au sein des échantillons traités. En outre, nous avons montré que le modèle ex vivo est comparable et corrélé au système de modèle de souris in vivo de référence pour l'essai de chimio-sensibilité. Suite à l’optimisation de ce modèle, nous avons collecté 25 échantillons de tumeurs de patientes atteintes de cancer de l’ovaire, pour réaliser des MDTs et des cultures de cellules primaires afin de comparer les profils transcriptomiques de ces deux modèles avec celui de la tumeur d’origine, et d'analyser les réponses aux traitements et le microenvironnement tumoral. Les données transcriptomiques obtenues par micropuces ARN nous ont permis d'effectuer une analyse bio-informatique des voies de signalisation incluant un groupement hiérarchique non supervisé. Nos résultats montrent que les MDT à chaque point de temps (jour 0, 8 et 15) sont génétiquement similaires à la tumeur primaire par opposition aux cultures cellulaires primaires, et que les principales voies dérégulées sont impliquées dans la réponse cellulaire au stress. Nous avons observé une viabilité élevée des cellules au sein des MDT sur une période de culture de 15 jours. En outre, nous avons déterminé qu'un régime de chimiothérapie (carboplatine et paclitaxel) consistant en une induction thérapeutique de 10 heures suivie d'une période de récupération de 14 heures était idéal pour caractériser la réponse au traitement. Notre analyse de prédiction de la réponse des patients montre que nous avons une corrélation positive élevée d'une efficacité de 95 % entre la réponse ex vivo et la réponse clinique pour les patients appariés. En général, nos résultats suggèrent que notre technique fournit un modèle plus sophistiqué et précis pour récapituler la réponse de la tumeur primaire dans un laps de temps cliniquement adapté, et pourrait servir de plateforme pour tester de nouvelles thérapeutiques, et d'outil d'orientation clinique pour la réponse des patients. / A major challenge in clinical oncology is the inability to accurately predict the patients’ response to therapeutic agents. Currently, there are no reliable models and assays available that reiterate the immense complexity of a primary tumor. These factors are important to implement a personalized medicine strategy capable of identifying the most suitable treatment regimen for a particular patient in a clinically relevant timeframe. To answer this need, our group has developed a novel ex vivo 3D model that relies on the micro-dissection of a patient’s tumor specimen and the utilization of microfluidic technologies to monitor drug sensitivity within a time-frame suitable for clinical decision-making. This approach would allow for better selection of effective therapies and limit the administration of ineffective treatments, further improving treatment outcome of patients while reducing cost and drug-induced toxicities. Previously published work studied that the viability of cancer cells located within the tumor was characterized using two imaging modalities: confocal microscopy and flow cytometry. However, each technique has its own disadvantage, limiting their ability to molecularly characterize the effect of therapeutic agents on cancer cells. Thus, we hypothesize that our 3D ex vivo tumor-derived model coupled to a pathology-like tool would allow for a more comprehensive approach to evaluate tumor response to treatment, providing a readout system to closely mirror the patient’s response, and evaluating molecular mechanisms involved in response to drugs. To address this hypothesis, we optimized a paraffin-embedding lithography procedure allowing the production of micro-dissected tissue micro-array (MDTMA) block to allow morphological and protein-specific staining to analyze the cellular integrity and tissue architecture of treated samples. In addition, we showed that ex vivo model is comparable and correlated to the gold standard in vivo mouse model system for chemosensitivity assay. Moreover, we collected, following informed consent, 25 post-surgical OC patient tumor samples, to form micro-dissected tissues (MDTs), and primary cell cultures for micro-array analysis and characterization of the TME and response prediction. The micro-array data allowed us to perform unsupervised hierarchical clustering and pathway analysis showing that the MDTs at each time-point (day 0, 8 and 15) are genetically similar to the primary tumor as opposed to the primary cell cultures and that main deregulated pathways are involved in cellular response to stress. We observed a high viability of cells within MDTs over a culture period of 15 days. In addition, we determined that a treatment regimen consisting of a 10-hour therapy induction followed by a 14-hour recovery period was ideal for characterizing carboplatin treatment response. Our response prediction analysis of patients shows that we have a high positive correlation of 95% efficiency between ex vivo and clinical response for matched patients. In general, our results suggest that our ex vivo drug response model provides a more sophisticated model to recapitulate primary tumor response in a clinically suitable timeframe that can be exploited further serving, in part, as a platform to test new therapeutics and as a clinical guidance tool for patient response.

The objectives of the study were (1) to quantify adherence rate among the study participants in the ART unit and (2) to identify factors that contribute to non-adherence. This cross sectional study was carried out at Nekemete referral clinic. Data was collected using a self-developed structured questionnaire where a total of 338 participants grouped into adherent and non-adherent based on a score derived from an adherence assessment were interviewed. Data was analysed using the Statistical Package for Social Sciences (SPSS) version 17.0. By using multivariate analysis of variables identified as correlates of adherence, non-adherence was common among those; with age between 18-30 yrs, with no education, who were not married, who had no pipe water supply, those with no electricity in the house, who perceived had no access to assistance from providers, who perceived the health care providers (HCPs) did not keep information confidentially, who had a language barrier with providers, and who were treated with a psychiatric illness. The study concludes that adherence is multi-factorial and varies significantly by individual and care setting. Psychosocial factors were found to impact adherence and should be analysed in more detail by further studies. Three psychosocial factors were independently associated with poor adherence: the study found that patients perceiving poor access; those perceiving problems in information confidentiality (and possibly experiencing stigmatisation); and having psychiatric morbidity (and possibly with less social support) are more likely to be non-adherent. Furthermore, individuals without electricity and those without piped water supply, implying low income, are at risk for non-adherence / Health Studies / M.A. (Public Health)

The objectives of the study were (1) to quantify adherence rate among the study participants in the ART unit and (2) to identify factors that contribute to non-adherence. This cross sectional study was carried out at Nekemete referral clinic. Data was collected using a self-developed structured questionnaire where a total of 338 participants grouped into adherent and non-adherent based on a score derived from an adherence assessment were interviewed. Data was analysed using the Statistical Package for Social Sciences (SPSS) version 17.0. By using multivariate analysis of variables identified as correlates of adherence, non-adherence was common among those; with age between 18-30 yrs, with no education, who were not married, who had no pipe water supply, those with no electricity in the house, who perceived had no access to assistance from providers, who perceived the health care providers (HCPs) did not keep information confidentially, who had a language barrier with providers, and who were treated with a psychiatric illness. The study concludes that adherence is multi-factorial and varies significantly by individual and care setting. Psychosocial factors were found to impact adherence and should be analysed in more detail by further studies. Three psychosocial factors were independently associated with poor adherence: the study found that patients perceiving poor access; those perceiving problems in information confidentiality (and possibly experiencing stigmatisation); and having psychiatric morbidity (and possibly with less social support) are more likely to be non-adherent. Furthermore, individuals without electricity and those without piped water supply, implying low income, are at risk for non-adherence / Health Studies / M.A. (Public Health)