Le cancer de la vessie est un cancer fréquent et extrêmement onéreux par patient puisque plusieurs patients subissent des récidives de cancer et ont recours parfois à des chirurgies complexes. Il est donc important de diagnostiquer efficacement ces cancers lors de la prise en charge initiale du patient. En effet, la procédure standard d’imagerie pour la détection du cancer est la cystoscopie de la vessie guidée par lumière blanche, toutefois cette méthode ne permet pas de bien distinguer les cellules qui sont propices à l’invasion musculaire des cellules de cancer de la vessie non infiltrant. Ce mémoire propose d’utiliser un nouvel immunoconjugué fluorescent ciblant la sous-unité alpha du récepteur de l’interleukine 5, un nouveau biomarqueur spécifique aux cellules du cancer de la vessie infiltrant, afin d’effectuer la cystoscopie de la vessie guidée par fluorescence. Pour ce faire, un protocole de conjugaison du fluorochrome cyanine-5 (Cy5) à un anticorps monoclonal a été développé. De plus, un protocole de conjugaison d’un peptide Cell Accumulator (ACCUM) sur cet anticorps fluorescent (A14-Cy5-ACCUM) a été optimisé. Ensuite, la capacité de cet immunoconjugué à marquer les cellules humaines de cancer de la vessie infiltrantes du muscle (MIBC), HT1376, a été testée. Par la suite, un nouveau modèle orthotpique murin de MIBC humain permettant la validation préclinique prochaine de l’A14-Cy5-ACCUM a été développé. Une banque de plasma et sérum sanguin, et d’urine de patients sains et atteints de cancer de la vessie a été compilé. Cette biobanque contient 111 échantillons de plasma sanguin et d’urine qui pourront être utilisé afin de tester l’hypothèse selon laquelle le niveau d’interleukine-5 sanguin pourrait être un facteur pronostique pour la progression du cancer de la vessie. Ce projet jette les bases pour l’évaluation potentielle de la cystoscopie guidée par fluorescence lors de la prise en charge initiale des patients atteints de cancer de la vessie afin d’améliorer la survie sans progression et la survie à long terme des patients atteints de MIBC. / Abstract: Bladder cancer is a frequent and extremely costly cancer when evaluated on a per-patient basis because of its high recurrence rate and patients undergoing complex medical procedures. It is of utmost importance to better identify the aggressiveness of this cancer at initial diagnosis. The standard procedure for bladder cancer detection is still white-light guided cystoscopy, which relies mostly on physicians experience in regard to identifying invasive malignancies. This memoir proposes the use of a new fluorescent immunoconjugate, targeting the alpha subunit of interleukin-5 receptor (IL-5R[apha]), a new biomarker specific to muscle-invasive bladder cancer (MIBC) cells for fluorescence-guided cystoscopy. To do so, a conjugation protocol to fluorescently label a monoclonal antibody with cyanine-5 fluorophores has been developped. Then, a conjugation protocol to attach Cell Accumulator (ACCUM) peptides to this fluorescent immunoconjugate (A14-Cy5-ACCUM) has been optimized. Moreover, the ability of A14-Cy5-ACCUM to stain MIBC cell line HT1376 has been tested. Most importantly, a novel orthotpic rat model of human MIBC for the future preclinical validation of fluorescence-guided cystoscopy in rat bladder has been developped. Finally, a new bladder cancer tissue repository at the CHUS has been established. This repository contains a total of 111 plasma and urine patient samples that will be helpful to evaluate if interleukin-5 blood levels could be used as a prognosis marker for bladder cancer progression. This project laid the basis for the potential evaluation of fluorescence-guided cystoscopy during initial diagnosis of bladder cancer patients to improve their disease-free and long-term survival.

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No. of bitstreams: 1 nunesneto_o_dr_bauru.pdf: 4276326 bytes, checksum: e73a2086ffde0d12d2f5875fb168f8c1 (MD5) Previous issue date: 2016-04-28 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Materiais e dispositivos baseados em compostos orgânicos desempenham um importante papel em diversas áreas da aplicação tecnológica devido às suas interessantes propriedades eletro-magneto- ópticas, adicionadas às suas características mecânicas únicas, facilidade de processamento, versatilidade de síntese e baixo custo relativo. Apesar do proeminente campo de aplicação destes materiais, muitos aspectos associados à sua ciência básica são ainda pouco compreendidos. Nesse cenário destaca-se o fenômeno de Magnetoresistência Orgânica (OMAR, da sigla em inglês). Tal fenômeno encontra-se associado a variações significativas da condutividade elétrica de dispositivos orgânicos induzidas por pequenos campos magnéticos externos em temperatura ambiente e tem sido observado em diversificados materiais poliméricos e moleculares. No presente trabalho avaliou-se o fenômeno de OMAR apresentado por um Diodo Emissor de Luz baseado na molécula de Alq3. Medidas de Espectroscopia de Impedância Elétrica na presença de um Campo Magnético estático externo (EIE-CM) foram realizadas sobre o referido dispositivo para diferentes temperaturas. Métodos diferenciados de aquisição e manipulação de dados foram empregados a fim de remover a dependência temporal dos sinais tipicamente observados. Os seguintes Efeitos de Campo Magnético (MFE, da sigla em inglês) foram observados sobre a resposta elétrica do dispositivo: (i) redução de cerca de 1% na resistência, efeito praticamente constante para todo o espectro de frequência e; (ii) variações significativas na capacitância, com intensificação do efeito de Capacitância Negativa em baixas frequências. Como suporte para a interpretação dos resultados experimentais foram realizadas simulações empregando-se duas abordagens: Circuitos Equivalentes e Análise de perturbações de pequenos sinais (em inglês, Small Signal Analysis ) via soluções numéricas das equações de transporte de Boltzmann numa aproximação por Drift-Diffusion empregando-se dispositivos simplificados. As análises sugerem que os MFE evidenciados podem estar associados a um aumento da mobilidade efetiva dos portadores de carga e a uma redução na taxa de recombinação bimolecular no dispositivo. Os resultados foram interpretados em termos dos modelos atualmente aceitos para o fenômeno de OMAR. Esta tese também apresenta um estudo de processos de geração e transferência de carga em corantes Cianinas, materiais promissores para aplicações em células solares com absorção no infravermelho. Técnicas de Ressonância de Spin Eletrônico induzida por Luz foram empregadas em blendas destes corantes com o polímero MEH-PPV e com o fulereno (C60) a fim de avaliar, respectivamente, o caráter aceitador e doador de elétrons das Cianinas. / Materials and devices based on organic compounds play an important role in various technological applications, mainly due to their interesting electrical-magneto-optical properties combined with their unique mechanical properties, easy processing, versatility of synthesis and relatively low cost. Despite the prominent application field of these materials many aspects associated with their basic science are still not well understood. In this context the Organic Magnetoresistance phenomenon (OMAR) deserves to be highlighted. This phenomenon is associated with significant changes in the electrical conductivity of organic devices induced by the presence of small external magnetic fields at room temperature, being observed in various polymeric and molecular materials. In this study we have investigated the OMAR phenomenon in Alq3-based OLEDs. Electrical impedance spectroscopy technique in the presence of an external static magnetic field (EIS-MF) was employed in the experiments; distinct temperatures were considered. Differentiated methods of acquisition and data manipulation were employed to remove the typically observed signal time dependence. The following magnetic field effects (MFE) were observed on the electrical response of the device: (i) a constant reduction of around 1% in the resistance over the entire frequency spectrum and; (ii) significant changes in the capacitance followed by an intensification of the negative capacitance effect at low frequencies. Simulations employing two different approaches were carried out for the interpretation of the experimental results: (i) Equivalent Circuits and (ii) Small Signal Analysis via numerical solutions of the Boltzmann transport equations by Drift-Diffusion approach. The results suggest that the observed MFE can be associated with an increase in the effective mobility of the charge carriers and a reduction in the bimolecular recombination rate in the device. The results were interpreted in terms of the currently accepted models for the OMAR phenomenon. This thesis also presents a study about generation and charge transfer processes in cyanine dyes (near infrared absorbing compounds) which are promising materials for applications in solar cells. Light induced Electron Spin Resonance (L-ESR) technique was employed to study the presence/formation of paramagnetic centers in blends of these dyes with MEH-PPV polymer and fullerene (C60) to evaluate, respectively, the electron acceptor and donor character of cyanine dyes. / FAPESP: 2011/21830-6 / CNPq: 204432/2013-8