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Biomarcadores Periféricos, Toxicidade Sistêmica E UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL INSTITUTO DE CIÊNCIAS BÁSICAS DA SAÚDE PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS BIOLÓGICAS: BIOQUÍMICA TESE DE DOUTORADO Biomarcadores periféricos, toxicidade sistêmica e regulação transcricional no transtorno bipolar: identificação de vias moleculares associadas com a sua fisiopatologia e potenciais alvos terapêuticos BIANCA PFAFFENSELLER Porto Alegre 2016 UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL INSTITUTO DE CIÊNCIAS BÁSICAS DA SAÚDE PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS BIOLÓGICAS: BIOQUÍMICA Biomarcadores periféricos, toxicidade sistêmica e regulação transcricional no transtorno bipolar: identificação de vias moleculares associadas com a sua fisiopatologia e potenciais alvos terapêuticos BIANCA PFAFFENSELLER Orientador: Prof. Dr. Fábio Klamt Coorientador: Prof. Dr. Flávio Kapczinski Tese apresentada ao Programa de Pós- Graduação em Ciências Biológicas: Bioquímica da Universidade Federal do Rio Grande do Sul, como requisito parcial à obtenção do grau de Doutor em Bioquímica. Porto Alegre 2016 CIP - Catalogação na Publicação Pfaffenseller, Bianca Biomarcadores periféricos, toxicidade sistêmica e regulação transcricional no transtorno bipolar: identificação de vias moleculares associadas com a sua fisiopatologia e potenciais alvos terapêuticos / Bianca Pfaffenseller. -- 2016. 248 f. Orientador: Fábio Klamt. Coorientador: Flávio Kapczinski. Tese (Doutorado) -- Universidade Federal do Rio Grande do Sul, Instituto de Ciências Básicas da Saúde, Programa de Pós-Graduação em Ciências Biológicas: Bioquímica, Porto Alegre, BR-RS, 2016. 1. transtorno bipolar. 2. EGR3. 3. regulação transcricional. 4. biomarcadores. 5. toxicidade. I. Klamt, Fábio, orient. II. Kapczinski, Flávio, coorient. III. Título. Elaborada pelo Sistema de Geração Automática de Ficha Catalográfica da UFRGS com os dados fornecidos pelo(a) autor(a). “Aprender é a única coisa de que a mente nunca se cansa, nunca tem medo e nunca se arrepende.” (Leonardo da Vinci) AGRADECIMENTOS Ao meu orientador, Professor Fábio Klamt, pela orientação, ensinamentos e dedicação a minha formação acadêmica e especialmente pela sua amizade e empolgação com a pesquisa. Ao meu coorientador, Professor Flávio Kapczinski, pela orientação e importante referência na área da pesquisa e pela oportunidade de aprendizado e crescimento profissional. Aos colegas e amigos do Laboratório de Psiquiatria Molecular, particularmente ao pessoal da bancada, com quem é muito bom compartilhar experiências e momentos divertidos no laboratório e fora dele. Destaco os amigos que estão no exterior, Gabriel, Gabi e Laura, que contribuíram muito na minha trajetória acadêmica. Em especial, agradeço à Bi Aguiar, minha xará, grande amiga, braço direito e parceria de sempre há mais de dez anos, com quem sempre pude contar. Um obrigado também muito especial à Gi, minha amiga-irmã, pelas trocas de ideias, apoio e consideráveis ajudas que foram fundamentais para a conclusão deste trabalho. Também agradeço à Bruna Maria, Emily e Thaís pelas conversas e parceria. E, mais recentemente, aos queridos amigos Ellen e Rafa Colombo. Pessoal, valeu pela amizade, companheirismo e pelo apoio de sempre. Aos demais colegas e professores do Laboratório de Psiquiatria Molecular, pela amizade e troca de experiência e conhecimentos, principalmente ao Pedro Magalhães pela importante contribuição neste trabalho. iii Aos amigos e colegas do Departamento de Bioquímica, principalmente à Pati, Dai, Ivi, Fer, que me auxiliaram no aprendizado e discussão de técnicas. E um obrigado essencial ao Marco Antônio pela sua disponibilidade, tranquilidade e por compartilhar seu conhecimento na área da bioinformática. Da mesma forma, agradeço ao Mauro Castro pela fundamental participação neste trabalho com seu admirável conhecimento e dedicação. As minhas grandes amigas da Biomed: Bianca, Roberta e Bárbara pela amizade e parceria desde a faculdade. À CAPES e ao CNPq pelas bolsas de estudo e financiamento de projetos. Ao Programa de Pós-graduação em Ciências Biológicas: Bioquímica pela excelência em educação e ensino. À Cleia pela simpatia, auxílio e esclarecimentos constantes. Um agradecimento mais que especial ao Adriano pela companhia, amor e carinho. Muito obrigada por me apoiar e compreender as minhas ausências e preocupações. A minha família, pela paciência, confiança, dedicação e compreensão em todos os momentos desta trajetória, principalmente naqueles em que estive mais ausente. Destaco meu pai Eliseu, que sempre incentivou o estudo e a busca pelo conhecimento. Agradeço por tudo! iv SUMÁRIO PARTE I ..................................................................................................................... 1 RESUMO .................................................................................................................... 2 ABSTRACT ................................................................................................................ 3 Lista de abreviaturas .................................................................................................. 4 1. INTRODUÇÃO ...................................................................................................... 7 1.1 Transtorno bipolar ............................................................................................... 7 1.2 Biomarcadores periféricos no transtorno bipolar .................................................. 9 1.2.1 Toxicidade sistêmica no transtorno bipolar ................................................. 11 1.3 Progressão do transtorno bipolar ........................................................................ 13 1.4 Resiliência e plasticidade celular ....................................................................... 14 1.5 Genética do transtorno bipolar ........................................................................... 16 1.6 Metodologias para estudo da genética em transtornos psiquiátricos.................... 17 1.6.1 Redes de regulação transcricional e análise de ‘reguladores mestres’ .......... 19 1.7 Modelos celulares para estudo do transtorno bipolar .......................................... 24 1.8 Justificativa ....................................................................................................... 26 2. OBJETIVOS ......................................................................................................... 27 2.1 Objetivo geral .................................................................................................... 27 2.2 Objetivos específicos ......................................................................................... 27 PARTE II .................................................................................................................. 29 3. ARTIGOS CIENTÍFICOS ................................................................................... 30 3.1 CAPÍTULO 1: Neurotrophins, inflammation and oxidative stress as illness activity biomarkers in bipolar disorder ...................................................................... 30 3.2 CAPÍTULO 2: Anatomical faces of neuroprogression in bipolar disorder......... 47 3.3 CAPÍTULO 3: Differential expression of transcriptional regulatory units in the prefrontal cortex of patients with bipolar disorder: potential role of early growth response gene 3 .......................................................................................................... 50 3.4 CAPÍTULO 4: Caracterização neuronal do modelo de células de neuroblastoma humano SH-SY5Y diferenciadas para estudo de alvos relacionados ao transtorno bipolar ....... 60 v 3.5 CAPÍTULO 5: Reduced Neurite Density in Neuronal Cell Cultures Exposed to Serum of Patients with Bipolar Disorder ................................................................................................................................... 97 PARTE III ............................................................................................................... 103 4. DISCUSSÃO ....................................................................................................... 104 5. CONCLUSÕES ................................................................................................... 124 6. PERSPECTIVAS ................................................................................................ 126 REFERÊNCIAS ...................................................................................................... 128 ANEXOS ................................................................................................................. 154 ANEXO 1: Artigo: Staging and neuroprogression in bipolar disorder ...................... 155 ANEXO 2: Capítulo de livro: Fisiopatologia do transtorno bipolar: novas tendências ................................................................................................................................. 164 ANEXO 3: Artigo: Early apoptosis in peripheral blood mononuclear cells from patients with bipolar disorder. ............................................................................................... 180 ANEXO 4: Artigo: Dissimilar Mechanism of Action and Dopamine Transporter Dependency of 6-Hydroxydopamine-Induced Toxicity in Undifferentiated and RA- Differentiated SH-SY5Y Human Neuroblastoma Cells. ............................................ 184 ANEXO 5: Lista de genes-alvo das unidades regulatórias enriquecidas no transtorno bipolar e seu modo de ação. ...................................................................................... 221 vi APRESENTAÇÃO A presente
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