Establishing a high titer transient gene expression process in conditioned media for CHO-DG44 cells

Dissertação para obtenção do Grau de Mestre em Biotecnologia

The role of Rac1-modulated gene transcription in tumorigenesis

Dissertation presented to obtain the Ph.D degree in Biology

Contribution to drug discovery and development for tauopathies using yeast as a model

This work aimed to contribute to drug discovery and development (DDD) for tauopathies, while expanding our knowledge on this group of neurodegenerative disorders, including Alzheimer’s disease (AD). Using yeast, a recognized model for neurodegeneration studies, useful models were produced for the study of tau interaction with beta-amyloid (Aβ), both AD hallmark proteins. The characterization of these models suggests that these proteins co-localize and that Aβ1-42, which is toxic to yeast, is involved in tau40 phosphorylation (Ser396/404) via the GSK-3β yeast orthologue, whereas tau seems to facilitate Aβ1-42 oligomerization. The mapping of tau’s interactome in yeast, achieved with a tau toxicity enhancer screen using the yeast deletion collection, provided a novel framework, composed of 31 genes, to identify new mechanisms associated with tau pathology, as well as to identify new drug targets or biomarkers. This genomic screen also allowed to select the yeast strain mir1Δ-tau40 for development of a new GPSD2TM drug discovery screening system. A library of unique 138 marine bacteria extracts, obtained from the Mid-Atlantic Ridge hydrothermal vents, was screened with mir1Δ-tau40. Three extracts were identified as suppressors of tau toxicity and constitute good starting points for DDD programs. mir1Δ strain was sensitive to tau toxicity, relating tau pathology with mitochondrial function. SLC25A3, the human homologue of MIR1, codes for the mitochondrial phosphate carrier protein (PiC). Resorting to iRNA, SLC25A3 expression was silenced in human neuroglioma cells, as a first step towards the engineering of a neural model for replicating the results obtained in yeast. This model is essential to understand the mechanisms of tau toxicity at the mitochondrial level and to validate PiC as a relevant drug target. The set of DDD tools here presented will foster the development of innovative and efficacious therapies, urgently needed to cope with tau-related disorders of high human and social-economic impact.

"Berlim : Bric-à-Brac : projeto de criação/investigação

Berlim e a sua paisagem sonora suscitam emoções diversas capazes de desencadear um processo criativo. As palavras que se seguem são a expressão de um projeto que se quis catalisador de um olhar muito pessoal sobre esta cidade. Tendo como base, na criação, a estética da colagem, este projeto materializa-se numa performance final onde uma atriz, uma bailarina e um trombonista dão corpo a diversas emoções. Ainda na criação, é importante salientar o papel das técnicas de síntese na busca de novas sonoridades e o uso de tecnologias da música na composição e edição musical. Está patente, neste projeto, um clin-d’oeil ao serialismo e a corrente espectral francesa. As anotações dramatúrgicas foram também essenciais ao longo da composição musical e da encenação de toda a performance.