Le cancer de la vessie représente un vrai problème de santé publique, avec une surveillance et suivi clinique à long terme en raison de l’importance des fréquences de récidives. La chimiothérapie reste souvent inefficace. L’objectif de cette thèse est donc la recherche de marqueurs sérologiques et moléculaires à valeur pronostique dans le cancer de la vessie qui peuvent servir à prédire la maladie. D’abord, nous avons étudié trois facteurs de transcriptions des lymphocytes T activées qui sont T-bet, GATA-3 et Bcl-6. Nous avons montré une surexpression de T-bet chez les malades à stade invasif et de haut grade, cependant, la surexpression de GATA-3 et Bcl-6 a été corrélée au stade superficiel et de bas grade. La survie a été corrélée avec le groupe des malades sans histoires de récidive ou progression et avec la surexpression de Bcl-6 et GATA-3. Cependant les malades qui expriment fortement T-bet répondent mieux au BCG. Ensuite, nous avons visé la détection de FasL et TRAIL solubles dans le sérum des malades atteints du cancer vésicale. Nous avons montré une surexpression de sFasL et sTRAIL chez les malades à stade superficiel et de bas grade. Le rôle anti-tumoral de ces cytokines a été confirmé sur deux lignées du cancer de la vessie montrant que le traitement avec le sérum riche en sFasL ou en sTRAIL diminue la viabilité cellulaire in vitro. A la fin de cette thèse, nous avons testé l’activation p-Akt dans la tumeur vésicale. Nous avons montré une surexpression de p-Akt au sein des tumeurs comparées au tissu sain adjacent, et au sein des malades à stade invasif et de haut grade. Akt semble être un marqueur de progression tumorale dans le cancer de la vessie. / Bladder cancer is the first most common urogenital cancer in men in Tunisia, with a high recurrence rate. Patients with muscle-invasive disease develop metastasis. The need for expensive continuous surveillance. In this thesis we try to search some candidate biomarkers. Their use for cancer staging and personalization of therapy at the time of diagnosis in order to identify a better treatment could improve patient care. The aim of this first part of our study was to investigate the clinical significance of three immune cell-related transcription factors, T-bet, GATA-3 and Bcl-6 in Tunisian patients with bladder cancer. We found that T-bet level was significantly higher in invasive carcinoma with high- grade. However, T-bet is predictive of response to BCG. On the contrary, the expression of GATA-3 and Bcl-6 was significantly higher in non-invasive carcinoma with low grade. We furthermore studied the effect of activation of soluble FasL and TRAIL molecule in bladder cancer. We demonstrate that the mean serum level of sFasL was higher in patients than in normal donors. sFasL was only higher than in sera of healthy donors where patients had superficial stage and low- and medium-grade cancer. sTRAIL was significantly lower in sera from patients with invasive and high-grade bladder carcinoma than in controls. Finally, we demonstrate that p-Akt levels in patients with invasive carcinoma and high-grade bladder cancer were significantly elevated compared to patients with non-invasive and low grade bladder cancer. Altogether, our results suggest that Akt activation can provide useful prognostic information.

La cystoscopie est actuellement l'examen clinique de référence permettant l'exploration visuelle des parois internes de la vessie. Le cystoscope (instrument utilisé pour cet examen) permet d'acquérir une séquence vidéo des parois épithéliales de la vessie. Cependant, chaque image de la séquence vidéo ne visualise qu'une surface réduite de quelques centimètres carrés de la paroi. Les travaux réalisés dans le cadre de cette thèse ont pour objectif de construire une carte 3D reproduisant d'une manière fidèle les formes et les textures des parois internes de la vessie. Une telle représentation de l'intérieur de la vessie permettrait d'améliorer l'interprétation des données acquises lors d'un examen cystoscopique. Pour atteindre cet objectif, un nouvel algorithme flexible est proposé pour le calibrage de systèmes cystoscopiques à vision active. Cet algorithme fournit les paramètres nécessaires à la reconstruction précise de points 3D sur la portion de surface imagée à chaque instant donné de la séquence vidéo cystoscopique. Ainsi, pour chaque acquisition de la séquence vidéo, un ensemble de quelques points 3D/2D et une image 2D est disponible. L'objectif du deuxième algorithme proposé dans cette thèse est de ramener l'ensemble des données obtenues pour une séquence dans un repère global pour générer un nuage de points 3D et une image panoramique 2D représentant respectivement la forme 3D et la texture de la totalité de la paroi imagée dans la séquence vidéo. Cette méthode de cartographie 3D permet l'estimation simultanée des transformations 3D rigides et 2D perspectives liant respectivement les positions du cystoscope et les images de paires d'acquisitions consécutives. Les résultats obtenus sur des fantômes réalistes de vessie montrent que ces algorithmes permettent de calculer des surfaces 3D reproduisant les formes à retrouver / Cystoscopy is currently the reference clinical examination for visual exploration of the inner walls of the bladder. A cystoscope (instrument used in this examination) allows for video acquisition of the bladder epithelium. Nonetheless, each frame of the video displays only a small area of few squared centimeters. This work aims to build 3D maps representing the 3D shape and the texture of the inner walls of the bladder. Such maps should improve and facilitate the interpretation of the cystoscopic data. To reach this purpose, a new flexible algorithm is proposed for the calibration of cystoscopic active vision systems. This algorithm provides the required parameters to achieve accurate reconstruction of 3D points on the surface part imaged at each given moment of the video cystoscopy. Thus, available data for each acquisition are a set of few 3D points (and their corresponding 2D projections) and a 2D image. The aim of the second algorithm described in this work is to place all the data obtained for a sequence in a global coordinate system to generate a 3D point cloud and a 2D panoramic image representing respectively the 3D shape and the texture of the bladder wall imaged in the video. This 3D cartography method allows for the simultaneous estimation of 3D rigid transformations and 2D perspective transformations. These transformations give respectively the link between cystoscope positions and between images of consecutive acquisitions. The results obtained on realistic bladder phantoms show that the proposed method generates 3D surfaces recovering the ground truth shapes

L'objectif de cette thèse est de faciliter le diagnostic du cancer de la vessie. Durant une cystoscopie, un endoscope est introduit dans la vessie pour explorer la paroi interne de l'organe qui est visualisée sur un écran. Cependant, le faible champ de vue de l'instrument complique le diagnostic et le suivi des lésions. Cette thèse présente des algorithmes pour la création de cartes bi- et tridimensionnelles à large champ de vue à partir de vidéo-séquences cystoscopiques. En utilisant les avancées récentes dans le domaine de la minimisation d'énergies discrètes, nous proposons des fonctions coût indépendantes des transformations géométriques requises pour recaler de façon robuste et précise des paires d'images avec un faible recouvrement spatial. Ces transformations sont requises pour construire des cartes lorsque des trajectoires d'images se croisent ou se superposent. Nos algorithmes détectent automatiquement de telles trajectoires et réalisent une correction globale de la position des images dans la carte. Finalement, un algorithme de minimisation d'énergie compense les faibles discontinuités de textures restantes et atténue les fortes variations d'illuminations de la scène. Ainsi, les cartes texturées sont uniquement construites avec les meilleures informations (couleurs et textures) pouvant être extraites des données redondantes des vidéo-séquences. Les algorithmes sont évalués quantitativement et qualitativement avec des fantômes réalistes et des données cliniques. Ces tests mettent en lumière la robustesse et la précision de nos algorithmes. La cohérence visuelle des cartes obtenues dépassent celles des méthodes de cartographie de la vessie de la littérature / The aim of this thesis is to facilitate bladder cancer diagnosis. The reference clinical examination is cystoscopy, where an endoscope, inserted into the bladder, allows to visually explore the organ's internal walls on a monitor. The main restriction is the small field of view (FOV) of the instrument, which complicates lesion diagnosis, follow-up and treatment traceability.In this thesis, we propose robust and accurate algorithms to create two- and three-dimensional large FOV maps from cystoscopic video-sequences. Based on recent advances in the field of discrete energy minimization, we propose transformation-invariant cost functions, which allow to robustly register image pairs, related by large viewpoint changes, with sub-pixel accuracy. The transformations linking such image pairs, which current state-of-the-art bladder image registration techniques are unable to robustly estimate, are required to construct maps with several overlapping image trajectories. We detect such overlapping trajectories automatically and perform non-linear global map correction. Finally, the proposed energy minimization based map compositing algorithm compensates small texture misalignments and attenuates strong exposure differences. The obtained textured maps are composed by a maximum of information/quality available from the redundant data of the video-sequence. We evaluate the proposed methods both quantitatively and qualitatively on realistic phantom and clinical data sets. The results demonstrate the robustness of the algorithms, and the obtained maps outperform state-of-the-art approaches in registration accuracy and global map coherence

Les variations en nombre de copies (CNV) sont des gains ou pertes d’une séquence d’ADN et peuvent avoir un rôle dans la susceptibilité à certaines maladies. Les CNVs peuvent être détectés par les puces de SNPs de haute résolution en analysant les intensités des allèles avec des algorithmes de détection des CNVs tels que CNV partition, PennCNV et QuantiSNP. Dans cette thèse, nous avons évalué les performances de ces outils pour la détection des CNVs au niveau pangénomique et pour les tests d'association. Nous avons également étudié des stratégies d'association combinant les informations de l'allèle et du nombre de copies pour des SNP situés dans des CNV. Nous avons appliqué ces outils pour mener une étude d’association pan-génomique avec les CNV en utilisant les données de l'étude espagnole du cancer de lavessie (SBC)/EPICURO générées par la puce Illumina 1M.Nos résultats montrent une faible fiabilité et une faible sensibilité des algorithmes de détection des CNV. Dans la région du gène GSTM1 où un CNV très fréquent existe qui est associé au risque de cancer de la vessie, nous avons constaté que les algorithmes de détection des CNV ont de faibles performances. Néanmoins, l’utilisation de la mesure d'intensité des allèles dans les tests d'association peut alors être une alternative intéressante car cela nous a permis de détecter cette association connue. Pour les SNPs situés dans des CNVs, nous avons étudié plusieurs stratégies de tests d'association et nous avons montré que la plus puissante était d’utiliser un modèle avec deux termes correspondant respectivement à la somme et à la différence du nombre de copies des deux allèles. Finalement, en appliquant ces stratégies à l'étude (SBC)/EPICURO, nous avons identifié des CNVs potentiellement associés au risque de cancer de la vessie, ainsi que des SNP dont l'allèle et le nombre de copies pourraient être impliqués dans le risque de cancer de la vessie. / Copy number variations (CNVs) are losses or gains of DNA sequences that may play a role in specific disease susceptibility. CNVs can be detected by high-resolution SNP-arrays through the analysis of allele intensities with CNV calling algorithms such as CNVpartition, PennCNV and QuantiSNP. In this thesis, we identified and assessed the performances of available tools for CNV calling and for association testing, at the genome-wide level. We also investigatedassociation strategies that combine information on both the allele and the number of copies for SNPs located in CNV regions. We applied these tools to conduct a genome-wide association study with CNV using data from the Spanish Bladder Cancer (SBC)/EPICURO Study generated by the Illumina 1M SNP-array. Our results showed a low reliability and a low sensitivity of the investigated CNV calling algorithms applied to SNP-array data. The GSTM1 locus shows a very frequent CNV that is associated with bladder cancer (BC) risk. We reported that the calling algorithms performed very poorly in identifying this CNV. We proposed using allele intensity measures (LRR) as a screening step to assess association as it allowed the detection of the GSTM1 CNV association with BC. To combine the allele and the number of copies for SNPs located in CNV regions, we investigated several strategies of association testing and we showed that the more powerfulone used a two-term model with the sum and the difference of the number of copies of both alleles. Finally, by applying these strategies to the (SBC)/EPICURO Study, we identified CNV regions potentially associated with BC risk, as well as SNPs for which both the allele and the number of copies could be involved in BC risk.

This thesis concerns the investigation of novel immunotherapies for cancer eradication. CpG therapy was used in order to target antigen-presenting cells (APCs), facilitating antigen presentation and activation of T cells. Blockade of the two major immune checkpoint regulators (CTLA-4 and PD-1) was also studied to ensure proper and sustained T cell activation. The therapies were investigated alone and compared to BCG, the standard immunotherapy in the clinic today for bladder cancer. In addition, CpG as well as BCG was combined with CTLA-4 or PD-1 blockade to examine if the combination could improve therapy. Single and combination strategies were assessed in an experimental bladder cancer model. In addition, one of the therapies (local aCTLA-4 administration) was evaluated in an experimental pancreatic cancer model. To be able to study the effects of CpG in humans, a human whole blood loop system has been used. This allowed us to dissect the potential interplay between CpG and complement. CpG was found to be superior to the conventional therapy, BCG, in our experimental model and T cells were required in order for effective therapy to occur. Used as a monotherapy, CTLA-4 blockade but not PD-1 blockade, prolonged survival of mice. When CTLA-4 or PD-1 blockade was combined with CpG, survival was enhanced and elevated levels of activated T cells were found in treated mice. In addition, Treg levels were decreased in the tumor area compared to tumors in control treated mice. CTLA-4 blockade was also effective when administrated locally, in proximity to the tumor. Compared to systemic CTLA-4 blockade, local administration gave less adverse events and sustained therapeutic success. When CpG was investigated in a human whole blood loop system it was found to tightly interact with complement proteins. This is an interesting finding which warrants further investigation into the role of TLRs in complement biology. Tumor therapy could be affected either negatively or positively by this interaction. The results presented herein are a foundation for incorporating these combination therapies into the clinic, specifically for bladder cancer but in a broader perspective, also for other solid tumors such as pancreatic cancer.

La present tesi avalua les diferències entre homes i dones quant a les característiques sociodemogràfiques i clínicopatològiques, procés diagnòstic, tractament i pronòstic en una sèrie de casos diagnosticats de novo de càncer de bufeta en 18 hospitals de 5 regions espanyoles (estudi EPICURO). També s'estudia l'associació entre tabac i risc de càncer de bufeta segons el sexe en un estudi agregat d'estudis cas-control europeus i nord-americans de càncer de bufeta.Es trobaren diferències entre sexes quant a la incidència de la malaltia, en algunes característiques anatomopatològiques dels tumors i quant a tractament. No es varen observar diferències entre sexes davant d'un mateix nivell d'exposició al tabac. També s'observaren diferències entre homes i dones quant al risc de recidivar i de progressar dels tumors vesicals superficials, però no en el risc de morir dels pacients amb tumors invasius. / The thesis evaluates differences related to sociodemographic and clinic-pathological characteristics, diagnostic tests, treatment and prognosis of bladder cancer patients newly diagnosed in 18 hospitals from 5 Spanish areas according to sex. The work also assess the association between tobacco and bladder cancer risk according to sex in a pooled analysis of case-control studies of bladder cancer from Europe and North America.Differences between sex concerning disease incidence, pathological characteristics and treatment were observed. The relative risk of bladder cancer associated with tobacco was similar in both sex. Differences between men and women were observed regarding risk of recurrence and progression of their superficial tumors but not regarding risk of death because of an invasive tumor.

L'objectiu d'aquest tesi és avaluar els diferents passos en el camí que va des de l'exposició a contaminants atmosfèrics/PAHs fins a la malaltia, el càncer de bufeta urinària. Es van mesurar partícules ultrafines a Barcelona. S'ha avaluat l'exposició a contaminació atmosfèrica en un estudi cas-control, recollint informació sobre la història residencial incloent diversos indicadors de l'exposició a contaminació atmosfèrica i altres factors de risc potencials. Es va dur a terme una revisió sistemàtica de la literatura per avaluar si els nivells de metabòlits del pirè i els aductes d'ADN i de proteïnes es correlacionaven amb nivells baixos d'exposició a PAHs. Vam mesurar els nivells d'aductes d'ADN en un subgrup d'individus de l'estudi cas-control amb la tècnica del radioetiquetatge amb fòsfor-32, tractament de la nucleasa P1. Vam analitzar 22 SNPs en set gens de la via de reparació de l'ADN per excisió de nucleòtids. / The aim of this thesis is to evaluate the different steps in the pathway from exposure (air-contaminants/PAHs) to disease (bladder cancer). We measured ultrafine particles in Barcelona. We evaluated the exposure to air pollutants in a case-control study, collecting information on the residential history with proxies for exposure to air pollution and other potential risk factors. We did a systematic review of the literature to evaluate if pyrene metabolites and DNA and protein adducts are correlated with low level exposure to PAHs. We measured bulky DNA adducts in a subgroup of subjects of the case-control study using 32P-Postlabeling, nuclease P1 treatment. We analyzed 22 SNPs in 7 genes of the nucleotide excision repair pathway.

Paclitaxel is one of the most successful drugs for the treatment of cancer because of its ability to target tubulin, block cell cycle progression at mitosis, and induce apoptosis. Despite the success of Paclitaxel, the development of drug resistance hampers its clinical applicability. Paclitaxel is used in malignant tumors in the present research, including non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC), head-neck scale epitheliomatous, urinary bladder cancer, tumor of the reproduction organ and so on. Clinical treatment of paclitaxel be injected 250mg/m2 to previously non-treated patients of small lung cancer its effect about 34% and previously non-treated patients of non-small lung cancer its effect about 21% to 24% . On the other hand we had established a taxol-resistant human lung carcinoma subline A549R by paclitaxel to compare the different proteins with A549 wild type and treat the different concention of paclitaxel with MTT assay so as to observe the tolerance dosage of subline growth.We obtained the patient¡¦s specimens of lung cancer to treat with paclitaxel some of resistant and some of non-resistant to compare differentially expressed proteins between normal and tumor. When we cultured taxol-resistant human lung carcinoma subline in which paclitaxel and calcium regulate growth, owing to the proteins of changes result to resistance. The extraction cell subline and specimens were analyzed by 2-D electrophoresis patterns and we found that interact paclitaxel with calcium it is the important factor of drug resistant. Verified from clinical treatment might have hypercalcemia in malignant tumors and calcium ion may increase paclitaxel drug resistance and hypercalcemia patient will be more insensitive to paclitaxel treatment.

During recent years, significant advances have been made in the field of molecular therapy in urologic oncology, mainly for advanced renal cell carcinoma. In this hitherto largely treatment-refractory disease, several agents have been developed targeting the von Hippel-Lindau metabolic pathway which is involved in carcinogenesis and progression of the majority of renal cell carcinomas. Although cure may not be expected, new drugs, such as the multikinase inhibitors sorafenib and sunitinib and the mammalian target of rapamycine inhibitor temsirolimus, frequently stabilize the disease course and may improve survival. Fewer data are available supporting molecular therapies in prostate, bladder, and testicular cancers. Preliminary data suggest a potential role of high-dose calcitriol and thalidomide in hormone-refractory prostate cancer, whereas targeted therapies in bladder and testicular cancers are still more or less limited to single-case experiences. The great theoretical potential and the multitude of possible targets and drug combinations, however, support further research into this exciting field of medical treatment of urologic malignancies. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

Background: In addition to conventional chemotherapeutics, nucleic acid-based therapeutics like antisense oligodeoxynucleotides (AS-ODN) represent a novel approach for the treatment of bladder cancer (BCa). An efcient delivery of AS-ODN to the urothelium and then into cancer cells might be achieved by the local application of multiwalled carbon nanotubes (MWCNT). In the present study, pristine MWCNT and MWCNT functionalized with hydrophilic moieties were synthesized and then investigated regarding their physicochemical characteristics, dispersibility, biocompatibility, cellular uptake and mucoadhesive properties. Finally, their binding capacity for AS-ODN via hybridization to carrier strand oligodeoxynucleotides (CS-ODN), which were either non-covalently adsorbed or covalently bound to the diferent MWCNT types, was evaluated. Results: Pristine MWCNT were successfully functionalized with hydrophilic moieties (MWCNT-OH, -COOH, -NH2, -SH), which led to an improved dispersibility and an enhanced dispersion stability. A viability assay revealed that MWCNTOH, MWCNT-NH2 and MWCNT-SH were most biocompatible. All MWCNT were internalized by BCa cells, whereupon the highest uptake was observed for MWCNT-OH with 40% of the cells showing an engulfment. Furthermore, all types of MWCNT could adhere to the urothelium of explanted mouse bladders, but the amount of the covered urothelial area was with 2–7% rather low. As indicated by fuorescence measurements, it was possible to attach CS-ODN by adsorption and covalent binding to functionalized MWCNT. Adsorption of CS-ODN to pristine MWCNT, MWCNT-COOH and MWCNT-NH2 as well as covalent coupling to MWCNT-NH2 and MWCNT-SH resulted in the best binding capacity and stability. Subsequently, therapeutic AS-ODN could be hybridized to and reversibly released from the CS-ODN coupled via both strategies to the functionalized MWCNT. The release of AS-ODN at experimental conditions (80 °C, bufer) was most efective from CS-ODN adsorbed to MWCNT-OH and MWCNT-NH2 as well as from CS-ODN covalently attached to MWCNT-COOH, MWCNT-NH2 and MWCNT-SH. Furthermore, we could exemplarily demonstrate that AS-ODN could be released following hybridization to CS-ODN adsorbed to MWCNT-OH at physiological settings (37 °C, urine). Conclusions: In conclusion, functionalized MWCNT might be used as nanotransporters in antisense therapy for the local treatment of BCa.