La leucémie myéloïde chronique (LMC) est un syndrome myéloprolifératif clonal initié par l’activité tyrosine kinase de l’oncoprotéine de fusion BCR-ABL dans une cellule souche hématopoïétique (CSH) très primitive et caractérisée par une instabilité génétique responsable de l’évolution clonale de la maladie. Les mécanismes de survie, d’autorenouvellement et ceux de la progression vers une phase de leucémie aigüe sont difficilement modélisables dans les CSH primaires de patients. La technologie des IPSC ; permettant de reprogrammer les cellules leucémiques à un état pluripotent ; pourrait permettre de générer in vitro des progéniteurs et des cellules leucémiques très primitives, dont l’évolution biologique pourrait être séquentiellement analysée. Dans ce but, nous avons généré une lignée IPSC à partir des cellules leucémiques d’un patient atteint de LMC. Nous avons montré que la lignée IPSC différenciée en hémangioblastes ou en progéniteurs hématopoïétiques présente un potentiel clonogénique accru. Ce potentiel est modulable sous l’action de l’imatinib mesylate ; qui inhibe l’autophosphorylation de BCR-ABL et celle de la protéine CRK-L ; mais également par la manipulation pharmacologique de la voie AHR impliquée dans la quiescence des CSH. En utilisant une stratégie de mutagénèse in vitro, nous avons montré la possibilté d’exacerber le potentiel hématopoietique des cellules hématopoïétiques dérivées des iPSC leucémiques. Les iPSC analysées après traitement par l’agent mutagène ENU présentent des anomalies cytologiques et cytogénétiques additionnelles reminiscentes de la phase blastiques de la maladie. Les analyses moléculaires par aCGH ont permis d’identifier dans les cellules hématopoïétiques dérivées d’IPSC traités par ENU, une signature moléculaire compatible avec celle décrite dans les cellules blastiques de patients. L’ensemble de nos résultats suggèrent qu’il est possible de modéliser certains aspects de la LMC ; notamment un phénotype myéloprolifératif ; et de génerer un modèle de progression blastique in vitro à partir des iPSC leucémiques, permettant ainsi d’identifier de nouveaux biomarquers prédictifs de progression tumorale ou le criblage de médicaments. / Chronic myeloid leukemia (CML) is a clonal myeloproliferative malignancy initiated by tyrosine kinase activity of the fusion oncoprotein BCR-ABL in very primitive hematopoietic stem cell and characterized by a genetic instability leading to clonal progression. Mechanisms of survival, self-renewal and progression of the disease are difficult to model using primary leukemic cells. The use of iPSC technology could allow reprogramming of leukemic cells to pluripotency with generation of primitive leukemic cells whose evolution can be sequentially analyzed. For this purpose, we generated an IPSC cell line from the leukemic cells of a CML patient and analyzed the possibility to generate a myeloproliferative phenotype. We have shown that this iPSC exhibits an increased hematopoietic potential either via EB or Blast-CFC generation. This potential can be modulated by the action of imatinib, inhibiting autophosphorylation of BCR-ABL and that of CRKL. We show that hematopoietic potential of CML iPSC can also be modulated by using AHR antagonists, which allow further amplification of hematopoietic cells. To evaluate the possibility of generating a clonal progression model in vitro, we have used a mutagenesis strategy. CML iPSC treated by ENU for several weeks generated hematopoietic cells with increased efficiency. These cells showed evidence of cytological and cytogenetic abnormalities reminiscent of a blast crisis. aCGH analyses of hematopoietic cells generated revealed genomic abnormalities described in CML blast crisis and a molecular signature compatible with blast crisis described in CML patients. These results suggest the feasibility of using patient specific iPSC for modeling CML blast crisis, which could be used for discovery of novel biomarkers and drug screening.

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2014-09-19T12:13:58Z No. of bitstreams: 2 Dissertacao Lucas C G Pereira.pdf: 869540 bytes, checksum: 9c40d119515c83b8b52606c52be6703d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-09-19T12:56:29Z (GMT) No. of bitstreams: 2 Dissertacao Lucas C G Pereira.pdf: 869540 bytes, checksum: 9c40d119515c83b8b52606c52be6703d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-09-19T12:56:29Z (GMT). No. of bitstreams: 2 Dissertacao Lucas C G Pereira.pdf: 869540 bytes, checksum: 9c40d119515c83b8b52606c52be6703d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-05-02 / In recent years, the evolution of health expenditures and specifically drugs, has worried governments. Among the various specialties, oncology is among those dealing with the greatest difficulties in the management of drug therapy. It is known that patients treated with various drugs have variability of response and susceptibility to drug toxicity. In present work, we study the role of Multiple Drug Resistance gene (MDR1) polymorphisms C1236T and C3435T frequencies and response to treatment with imatinib mesylate in 96 patients with chronic myeloid leukemia (CML). A total of 96 patients with CML were treated according to the Brazilian National Cancer Institute (INCA) guidelines and the blood samples were collected for genotyping. Genomic DNA was extracted and C1236T and C3435T polymorphisms genotyping was performed by the polymerase chain reaction with restriction fragment length polymorphism (PCR-RFLP), which detects a variation in length of a DNA fragment generated (370pb and 340pb) by a specific endonuclease in a specific site of the genome (HaeIII and MboI). Of the 96 CML samples, 31 samples were homozygous (CC), 13 homozygous (TT) and 52 heterozygous (CT) for exon 12 (1236). For the exon 26 (3435), 35 were homozygous (CC), 12 homozygous (TT) and 49 heterozygotes (CT). All frequencies for both polymorphisms were in Hardy-Weinberg equilibrium (p = 0.229 and q = 0.414). We found percentage association between polymorphisms and their distribution in different populations, and the response to treatment both cytogenetic and molecular difference was not statistically significant (p <0.05) when compared to age and sex presented response by patients and also no statistical difference (p <0,050). We conclude that the observed allele frequency for exons 1236 were 59.4% for C and 40.6% for T and the frequencies for the exon 3435 were 62.0% for C and 38.0% for T. That the relationship between the frequencies of polymorphisms of MDR1 in populations of different geographic locations, can provide tools that help in choosing a more appropriate and effective treatment of CML. / Neste estudo, o papel dos polimorfismos C1236T e C3435T do gene de Resistência a Múltiplas Drogas (MDR1) foram investigados em relação à frequência e a resposta ao tratamento com imatinibe em pacientes com leucemia mielóide crônica (LMC). Um total de 96 pacientes com LMC foram tratados de acordo com as diretrizes do Instituto Nacional do Câncer (INCA) e amostras de sangue foram coletadas para genotipagem do gene MDR (Resistência à Multiplas Drogas). O DNA genômico foi extraído e a genotipagem dos polimorfismos C1236T e C3435T foi realizada por meio da reação em cadeia da polimerase com fragmentos de restrição (PCR-RFLP), que detectou uma variação no comprimento de um fragmento de DNA gerado (370pb e 340pb) por uma endonuclease específica em um sítio específico do genoma (HaeIII e Mbol). Analisando as 96 amostras de pacientes para o polimorfismo no éxon 12 (1236) com LMC, 31 amostras apresentaram homozigose (CC), 13 homozigose (TT) e 52 heterozigose (CT). Para o estudo do polimorfismo no éxon 26 (3435), 35 foram homozigotas (CC), 12 homozigotas (TT) e 49 heterozigotas (CT). Todas as frequências para ambos os polimorfismos apresentaram-se em equilíbrio de Hardy-Weinberg (p = 0,229 e q = 0,414). Foi encontrada associação do percentual dos polimorfismos estudados em relação à distribuição dos mesmos em grupos de diferentes localizações geográficas, e sobre a resposta ao tratamento tanto citogenética e molecular, não houve diferença estatísticamente significante (p<0,05), quando foi comparado à idade e ao gênero apresentados pelos pacientes e a resposta também não houve diferença estatística (p<0,05). Conclui-se que as frequências alélicas observadas para o éxon 1236 foram de 59,4% para C e 40,6% para T e as frequências para o éxon 3435 foram de 62,0% para C e 38,0% para T e que a relação entre as frequências de polimorfismos de MDR1 nas populações de diferentes localizações geográficas, pode fornecer ferramentas que auxiliem na escolha de um tratamento mais adequado e eficaz da LMC.

The collapse of a stellar core is expected to produce gravitational waves (GWs), neutrinos, and in most cases a luminous supernova. Sometimes, however, the optical event could be significantly less luminous than a supernova and a direct collapse to a black hole, where the star just disappears, is possible. The GW event GW150914 was detected by the LIGO Virgo Collaboration via a burst analysis that gave localization contours enclosing the Large Magellanic Cloud (LMC). Shortly thereafter, we used DECam to observe 102 deg(2) of the localization area, including 38 deg(2) on the LMC for a missing supergiant search. We construct a complete catalog of LMC luminous red supergiants, the best candidates to undergo invisible core collapse, and collected catalogs of other candidates: less luminous red supergiants, yellow supergiants, blue supergiants, luminous blue variable stars, and Wolf-Rayet stars. Of the objects in the imaging region, all are recovered in the images. The timescale for stellar disappearance is set by the free-fall time, which is a function of the stellar radius. Our observations at 4 and 13 days after the event result in a search sensitive to objects of up to about 200 solar radii. We conclude that it is unlikely that GW150914 was caused by the core collapse of a relatively compact supergiant in the LMC, consistent with the LIGO Collaboration analyses of the gravitational waveform as best interpreted as a high mass binary black hole merger. We discuss how to generalize this search for future very nearby core-collapse candidates.

This thesis presents the characterization and calibration of the MOPITT-A instrument which uses the technique of correlation spectroscopy to ensure carbon monoxide in the atmosphere. A theoretical model is developed for the instrument and compared to MOPITT-A measurements collected under controlled laboratory conditions, which were designed to emulate atmospheric signals. It is shown that the model and measurements are in very good agreement with each other and that the MOPITT-A instrument behaves as expected. It was found that the gain of the instrument varies with time. The cause of the gain variation is not known but it is suggested that frosting inside the detector nest would be consistent with the observed nature of the variation.

This thesis presents the characterization and calibration of the MOPITT-A instrument which uses the technique of correlation spectroscopy to ensure carbon monoxide in the atmosphere. A theoretical model is developed for the instrument and compared to MOPITT-A measurements collected under controlled laboratory conditions, which were designed to emulate atmospheric signals. It is shown that the model and measurements are in very good agreement with each other and that the MOPITT-A instrument behaves as expected. It was found that the gain of the instrument varies with time. The cause of the gain variation is not known but it is suggested that frosting inside the detector nest would be consistent with the observed nature of the variation.

Les ressources étant limitées, les organismes doivent les partager entre les différents traits associés à leur valeur sélective. L'investissement optimal dans chacun de ces traits dépend des conditions environnementales et les individus sont sélectionnés pour ajuster leur stratégie d'allocation en conséquence. La théorie de la compétition locale pour l'accouplement (LMC) illustre bien cette hypothèse. Cette théorie prédit que lorsque qu'une population est structurée, de sorte de l'accouplement se fait localement et que des mâles apparentés sont en compétition pour la reproduction, les femelles devraient biaiser leur sexe ratio en faveur des filles. Bien que cette théorie ait été soutenue par de nombreuses études empiriques, les processus mécanistiques et évolutifs sous-jacents sont très peu connus. Dans cette thèse, j'aborde ces processus chez l'acarien haplo-diploïdeTetranychus urticae. Dans un premier temps, je montre que chez cette espèce, la taille des œufs es tutilisée par les femelles comme mécanisme d'ajustement du sexe-ratio. En effet, les œufs plus gros ont une plus grande probabilité de devenir femelle, et la taille des œufs produits augmente avec l'intensité de la LMC. Dans un second temps, à travers une approche d'évolution expérimentale, j'apporte le premier exemple empirique d'évolution du sexe-ratio en réponse à la LMC.En plus de varier avec les conditions environnementales, la stratégie optimale d'allocation desressources diffère souvent entre mâles et femelles, générant des conflits sexuels. Dans cette thèse,j'étudie les conséquences de tels conflits sur la stratégie reproductive des femelles chez Tetranychusurticae. Je montre notamment que l'accouplement induit une réduction de la longévité et de la fécondité, accompagnée d'une augmentation de la taille des œufs. / Because resources are limited, organisms are constrained to divide their resources between fitness components. The optimal investment into these traits can vary with environmental conditions, so that individuals are selected to adjust their allocation strategy accordingly. The theory of local mate competition (LMC) illustrates this hypothesis. This theory predicts that when populations arestructured, so that mating takes place locally and related males compete for mates, mothers should biastheir sex ratio towards females. Although this theory has been supported by many empirical studies,mechanistic and evolutionary processes underlying sex ratio adjustment remain poorly understood. Inthis thesis, I address these issues, using the haplodiploid spider mite Tetranychus urticae. First, I show that in this species, egg size is used as a mechanism of sex ratio adjustment. Indeed, larger eggs aremore likely to become female, and female spider mites use this property to adjust their sex ratio inresponse to LMC, increasing egg size with the intensity of LMC. Second, through an experimentalevolution approach, I provide the first empirical evidence that the sex ratio can evolve in response toLMC.In addition to varying with environmental conditions, the optimal strategy of resourceallocation also generally differs between male and females. Because of such conflicts, males havedeveloped adaptations aimed at manipulating the way by which females allocate their resources. Iinvestigate the consequences of mating for female reproductive strategies in spider mites. I find that mating induces a decrease of both longevity and fecundity, and an increase of egg size.

La leucémie myéloïde chronique (LMC) est une affection hématologique maligne caractérisée par l’apparition dans la moelle osseuse et le sang, d’un nombre important de globules blancs dont certains sont immatures. A l’heure actuelle, le principal traitement est l’Imatinib, commercialisé sous le nom de Glivec®. Ce traitement conduit, chez certains patients, à l'émergence de souches résistantes, ce qui complique la thérapeutique et nécessite un large panel de molécules actives pour contourner les mécanismes de résistance. Par conséquent, la recherche de nouvelles molécules bioactives agissant sur de nouvelles cibles biologiques reste toujours un défi important et d’actualité. Au cours de ce projet de thèse, nous nous sommes intéressés au développement de nouveaux procédés de synthèse pour accéder et découvrir de nouvelles molécules bioactives, en série nucléosidique, pour contourner les mécanismes de résistances dans des modèles de LMC. Nous avons tout d’abord développé une nouvelle méthode de synthèse propre et efficace de nouveaux analogues de C-nucléosides en utilisant une catalyse par le fer. Nous avons ensuite réalisé plusieurs modifications post-synthétiques pour accéder à de nouveaux C-nucléosides hautement fonctionnalisés, en particulier les analogues de la thiophènefurine. Nous avons également développé un nouveau procédé de synthèse d’une famille de O-glycosides, en combinant l’activation par les ultrasons à la catalyse par le fer. Ce procédé a été exploité par la suite pour concevoir et synthétiser les analogues glycosidiques du résvératrol. Enfin, nous avons mis au point une méthode efficace et peu couteuse d’azidation de sucres protégés. / Chronic myeloid leukemia (CML) is a malignant hematological disorder characterized by the formation, in the blood and bone marrow, of an excessive number of white blood cells, some of which are immature. Currently, the main treatment of CML is based on tyrosine-kinase inhibitors, such as Imatinib, the first approved drug of this class of compounds and marketed as Gleevec ®. However, this treatment results, in some patients, to the emergence of resistance, which complicates the treatment and requires a wide range of active molecules to circumvent resistance mechanisms. Therefore, the search for new bioactive molecules featuring new mode of action still remains a challenging.In this thesis project, we were interested in the design and discovery of new bioactive molecules in nucleoside series to circumvent resistance mechanisms in CML models, as well as in the development of new synthetic methods for the preparation of these targeted molecules. We first developed a clean and efficient procedure for the synthesis of new C- nucleoside analogues using iron catalysis. Then, several post-synthetic modifications were carried out, starting from halogenated nucleoside derivatives, to access highly functionalized C- nucleosides, as new analogues of thiophenefurin . We also developed a new procedure for the synthesis of O-glycosides using a cooperative effect of iron catalysis and ultrasound activation. This method has been applied for the synthesis of resveratrol O-glycosides. Finally, we developed an effecient and inexpensive method for anomeric sugar azidation. This method was applied for the synthesis of 1,2,3-triazolyl glycosides using a one-pot azidation-click procedure.

Orientador: Maria Aparecida Custódio Domingues / Resumo: Introdução: A Leucemia Mieloide Crônica (LMC) consiste em uma neoplasia hematopoiética maligna de células-tronco, caracterizada pela aquisição do gene de fusão BCR-ABL1 associado à translocação [t(9;22) (q34;q11)]. O início é insidioso com progressão lenta seguida por uma fase acelerada, que culmina na crise blástica. Os nichos da medula óssea estão relacionados com o microambiente tumoral, afetando a sobrevivência e a proliferação das células-tronco leucêmicas. Objetivos: Análise da quantidade e distribuição de células-tronco hematopoiéticas nos diferentes nichos na LMC em diferentes fases e correlação destes achados com a distribuição de galectina-3 nas fases acelerada e blástica. Material e métodos: Revisão clínico-morfológica de 20 pacientes na fase crônica da LMC, 1 paciente na fase acelerada e 1 paciente na crise blástica. Análise imunofenotípica (CD34, CD31, CD44, CD117 e galectina-3) em Tissue Microarrays para avaliação dos diferentes nichos e sua correlação com a matriz proteica. Resultados: Observou-se aumento na expressão de todos os marcadores na fase crônica quando comparados ao controle. Na fase acelerada há aumento na expressão de CD44 e galectina-3 quando comparado ao controle enquanto que na crise blástica há aumento na expressão de CD44 e CD117. Conclusão: A LMC constitui uma neoplasia mieloproliferativa em que o remodelamento dos nichos malignos favorece a manutenção das células-tronco leucêmicas. O presente estudo demonstrou elevação em todos os marcadores... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: Chronic Myeloid Leukemia (CML) is a malignant hematopoietic stem cell neoplasm, characterized by the acquisition of the translocation-associated BCR-ABL1 gene [t(9;22)(q34;q11)]. The onset is insidious with the slow progression followed by an accelerated phase, culminating in the blast crisis. The bone marrow niches are related to the tumor microenvironment, the survival and proliferation of the leukemic stem cells. Objectives: To analyze the amount and distribution of hematopoietic stem cells in different niches in CML in different phases and correlate these findings with the galectin-3 distribution in the accelerated and blastic phases. Material and methods: Clinical-morphological review of 20 patients in CML chronic phase, 1 patient in the accelerated phase and 1 patient in the blast crisis. Immunophenotypic analysis (CD34, CD31, CD44 and galectin-3) in Tissue Microarrays to evaluate the different niches and correlation with the protein matrix. Results: We observed the presence of all markers in the chronic phase when compared to the control. In the accelerated phase there is an increase in expression of CD44 and galectin-3, when compared to control while in blast crisis there is an increase in expression of CD44 and CD117. Conclusion: CML is a myeloproliferative neoplasm in which remodeling of malignant niches favors the maintenance of leukemic stem cells. The present study demonstrated an increase in expression of all immunohistochemical markers at differen... (Complete abstract click electronic access below) / Mestre

Red Supergiant (RSG) stars are the most luminous stars in the infrared sky. Their intrinsic luminosities combined with the low dust extinction observed in this regime makes these objects very attractive to study in the near-infrared (IR). In addition, RSGs are necessarily young objects, as they are tracers of recent star formation in extra-galactic systems. As the next generation of telescopes will be optimised for study in the near-IR, it is clear that, in the coming years, RSGs will play a prominent role in the way that astronomers probe the local Universe and out to larger distances with space-based observations. Therefore, it is vital to better our understanding of these objects now and develop the tools that will allow us to take full advantage of the suite of instrumentation that will become available in the near future. This thesis aims to further the understanding of RSGs by focusing on quantitative studies of near-IR spectroscopic observations. To this end, I develop an analysis technique that uses spectroscopic and photometric observations to estimate stellar parameters of RSGs. The observations are compared with synthetic spectra extracted from stellar model atmospheres, where departures from local thermodynamic equilibrium have been calculated for the diagnostic spectral lines. This technique is tested thoroughly on synthetic and real observations and is shown to reliably estimate stellar parameters in both regimes when compared with input parameters and previous studies respectively. Using the analysis routines developed in Chapter 3, in Chapter 4 I measure the chemistry and kinematics of NGC2100, a young massive cluster (YMC) of stars in the Large Magellanic Cloud, using near-IR spectroscopic observations of 14 RSGs taken with the new K-band multi-object spectrograph (KMOS). I estimate the average metallicity to be -0.43±0.10 dex, which is in good agreement with previous studies. I compare the observed location of the target RSGs on the Hertzsprung{Russell diagram with that of a Solar-like metallicity YMC and show that there appears to be no significant difference in the appearance of the RSGs in these two clusters. By combining the individual RSG spectra, I create an integrated-light cluster spectrum and show that the stellar parameters estimated, using the same technique as for individual RSGs, are in good agreement with the average properties of the cluster. In addition, I measure - for the first time - an upper limit of the dynamical mass of NGC2100 to be 15.2 X 10⁴Mʘ, which is consistent with the literature measurement of the photometric mass of the cluster. In Chapter 5, I present observations of RSGs in NGC6822, a dwarf irregular with a turbulent history, observed with KMOS. The data reduction process with KMOS is described in detail, in particular where the reduction has been optimised for the data. Stellar parameters are estimated using the technique presented in Chapter 3 and an average metallicity in NGC6822 of -0.55±0.13 dex is found, consistent with previous measurements of young stars in this galaxy. The spatial distribution of metallicity is estimated and weak evidence is found for a radial metallicity gradient, which will require follow-up observations. In addition, I show that the metallicities of the young and old populations of NGC6822 are well explained using a simple closed-box chemical evolution model, an interesting result, as NGC6822 is expected to have undergone significant recent interactions. In Chapter 6, I present multi-epoch KMOS observations of 22 RSGs in the Sculptor Group galaxy NGC55. Radial velocities are measured for the sample and are shown to be in good agreement with previous studies. Using the multi-epoch data, I find no evidence for radial velocity variables within the sample. Stellar parameters are estimated for 10 targets and are shown to be in good agreement with previous estimates. I conclude this thesis by summarising the main results and present a first-look calibration of the relationship between galaxy mass and metallicity using RSGs. By comparing the RSG metallicity estimates to metallicities estimated from ~ 50 000 Sloan digital sky survey galaxies, I show that the absolute metallicities of the two samples disagree. A more quantitative analysis requires additional RSG observations. In addition, using ~ 80 RSGs, with stellar parameters estimated in a consistent way, I show that there appears to be no dependence of the temperature of RSGs upon metallicity. This is in disagreement with current evolutionary models, which display a temperature change of ~ 450K over the studied range in metallicity. Finally, I outline potential areas for future work, focusing on follow-up studies that have been identified as a result of the work done in this thesis.

La leucémie myéloïde chronique (LMC) est un syndrome myéloprolifératif lié à l’acquisition d’une anomalie chromosomique t(9;22) conduisant à l’expression d’une protéine de fusion p210 Bcr-Abl dont l’activité tyrosine kinase dérégulée est nécessaire et suffisante pour engendrer la maladie.Cette pathologie bénéficie depuis 2002 d’une avancée thérapeutique : les inhibiteurs de tyrosine kinase (ITK). Cette thérapeutique dite ciblée, dont le chef de file est l’imatinib, est très efficace puisque 80% des patients entre en rémission. Malheureusement, 20% des patients traités développent des résistances primaires ou secondaires, dépendantes ou non de l’oncogène Bcr-Abl dont certaines ont été caractérisées. A ce titre, la LMC est devenue un modèle d’étude à la fois des mécanismes oncogéniques mais aussi des résistances.La résistance aux ITK dans la LMC peut être considérée sur deux plans. D’une part la résistance qui permet à la cellule leucémique d’échapper à la pression thérapeutique des ITK et d’autre part la résistance « intrinsèque » de la cellule souche leucémique par des mécanismes certainement multiples. Ce second niveau de résistance est à l’origine de la récurrence de la LMC lors de l’arrêt du traitement.Cette thèse a consisté à déterminer comment pouvait mourir les cellules de LMC en réponse aux ITK pour mettre en évidence les morts induites et les régulations qui existent entre-elles. De plus, cela a permis d’utiliser les morts non-apoptotiques pour contourner les mécanismes de résistance aux ITK.Nous avons montré pour la première fois en utilisant différents modèles cellulaires de LMC (cellules K562, Lama-84 et AR-230), que l’imatinib (ainsi que les autres ITK nilotinib et dasatinib) induit de la sénescence en plus d’une réponse apoptotique. En absence d’apoptose, par inhibition de cette dernière, la réponse sénescente devient une réponse majeure des cellules de LMC suggérant que l’apoptose a un rôle de « frein » sur la sénescence. L’autophagie activée par les ITK régule négativement la réponse apoptotique alors qu’elle est nécessaire pour une réponse sénescente majeure. Nous avons pu mettre en évidence deux types de sénescences induites par l’imatinib : une sénescence dépendante et une indépendante de l’autophagie. L’autophagie semble donc au cœur de la régulation des morts cellulaires. Puisque les cellules de LMC peuvent mourir par des morts non-apoptotiques, nous avons cherché à éliminer les cellules résistantes par des morts non-apoptotiques. Pour cela différentes molécules ont été utilisées telles que l’acide mycophénolique (MPA), un immunosuppresseur déjà utilisé en clinique. Le MPA en inhibant la synthèse de GTP permet d’induire des dommages à l’ADN et une réponse apoptotique et/ou sénescente. Dans ce contexte, l’autophagie protège la cellule de la réponse apoptotique mais ne protège pas la cellule de la sénescence. Le MPA est au contraire un puissant inducteur d’apoptose sur les cellules primaires. En effet, il induit une apoptose massive des cellules primaires résistantes aux ITK quelque soit le mécanisme impliqué (surexpression de tyrosine kinase, mutation de Bcr-Abl). Le MPA est l’exemple parfait des molécules qu’il nous faut rechercher pour éliminer les cellules résistantes de LMC notamment dans le cas où les patients sont en crise blastique et donc résistants aux thérapeutiques.Ces résultats suggèrent que la sénescence est une des morts qui peut être induite pour dépasser la résistance des cellules cancéreuses. / Chronic Myeloid Leukemia is a myeloproliferative syndrome connected to the acquisition of a chromosomal abnormality t(9;22) leading to the expression of a fusion protein p210 Bcr-Abl of whom the tyrosine kinase activity deregulated is necessary and sufficient to engender the disease.This pathology benefits since 2002 of a therapeutic advance: the tyrosine kinase inhibitors (TKI). This targeted therapeutics, from which imatinib is the front-line, is very effective because 80 % of patients enters in remission. However, 20 % of the treated patients develop primary or secondary resistances which can be dependent or not to the Bcr-Abl oncogene among which some have been characterized. Indeed, CML is now a model to study both oncogenic and resistances mechanisms.Resistance to TKI in CML can be considered on two sides. On one hand the resistance allowing the leukemic cell to escape the therapeutic pressure of TKI and on a second hand the “intrinsic” resistance of Leukemic stem cells by multiple mechanisms. This second level of resistance is at the origin of the CML recurrence.This thesis consisted in determining how could die the CML cells in response to TKI to bring to light cell deaths induced and the regulations existing between them. Furthermore, it allowed exploring the use of non-apoptotic cell deaths to overcome resistance to TKI.We showed for the first time by using CML cell lines (K562, Lama-84 and AR-230), that imatinib (as well as nilotinib and dasatinib) induced senescence besides an apoptotic response. In absence of apoptosis, by its inhibition, senescence becomes a major response of CML cells suggesting that apoptosis is limiting senescence. Autophagy activated by TKI negatively regulates apoptosis while it is necessary for a major senescent response. We were able to bring to light two types of senescence in response to TKI : a senescence dependent and a senescence independent of autophagy suggesting it plays a critical role in cell death regulation.Because CML cells can die by non-apoptotic cell deaths, we used them to eliminate TKI resistant cells. Mycophenolic acid (MPA), an immonusuppressor already used in therapeutic as an immunosuppressive agent has been extensively used. MPA by inhibiting the synthesis of GTP induces DNA damage and apoptotic and\or senescent response. In this context, autophagy protects the cells from apoptotic response but do not from senescence. Conversely, MPA is a powerful inductor of apoptosis on hematopoietic primary cells. Indeed, it induces apoptosis of TKI resistant primary cells whatever the mechanism involved (overexpression of tyrosine kinases or mutation of Bcr-Abl). MPA illustrates the need to look for new molecules to eliminate TKI resistant CML cells, particularly when patients are in the evolved blastic phase of the disease.These results suggest that senescence is one of the deaths which can be used to overcome resistance of cancer cells.