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Melanoma is the deadliest form of skin cancer. Prognosis for early stage melanoma patients is excellent, and surgery is often curative for these patients. However, once patients have presented with invasive disease, the average 5-year survival rate drops significantly from over 90% to between 10 and 15%. Several therapies have been developed to target a commonly mutated oncogene BRAF, or its downstream effectors. Unfortunately, while these treatments show robust initial response, most patients relapse within a year. Moreover, therapy-resistant tumors are often more invasive and metastatic. Therefore, it is important to investigate the molecular mechanisms underlying melanoma invasion and metastasis, and to prevent melanoma cell dissemination and metastatic progression. Invadopodia are proteolytic membrane protrusions used by metastatic cancer cells to degrade the extracellular matrix and to facilitate cancer cell invasion and metastasis. In my thesis research I have focused on protein fucosylation and store-operated calcium entry, two separate mechanisms involved in invadopodial regulation. Post translational modifications of proteins are essential for their structure and function. Many cell surface proteins require modifications such as glycosylation for protein-protein interactions, cell adhesion, and signal transduction. Fucosylation is a form of glycosylation that adds L-fucose on glycan structures of proteins. There is evidence indicating that fucosylation plays an important but cancer-type and branching dependent role in cancer progression. Emerging evidence indicates that the fucose salvage pathway and protein fucosylation are altered during melanoma progression and metastasis. Here, we report that the fucose salvage pathway inhibits invadopodia formation and extracellular matrix degradation by promoting α(1,2) fucosylation of cell surface proteins. The activation of the fucose salvage pathway decreases invadopodia numbers and inhibits the proteolytic activity of invadopodia in WM793 melanoma cells. Inhibiting fucokinase, one of the critical enzymes in the fucose salvage pathway, in melanoma cells abrogates L-fucose-mediated inhibition of invadopodia, suggesting dependence on the fucose salvage pathway. The inhibition of invadopodia formation by L-Fucose treatment or fucokinase overexpression could be rescued by treatment with α(1,2), but not α(1,3/4) fucosidase, implicating an α(1,2) fucose linkage-dependent inhibitory effect. The ectopic expression of FUT1, an α(1,2) fucosyltransferase, is sufficient to inhibit invadopodia formation and ECM degradation. Our findings indicate that the fucose salvage pathway can inhibit invadopodia formation, and consequently, invasiveness in melanoma via α(1,2) fucosylation. Re-activation of this pathway in melanoma could be useful for preventing melanoma invasion and metastasis. Calcium is a critical second messenger involved in a multitude of biological processes from cell proliferation to muscle contraction. In melanoma, previous studies have found that activation of the store operated calcium entry (SOCE) channel promotes tumor invasion and metastasis, in vitro and in xenograft models. The expression levels of STIM1, an essential component of the store operated calcium channels, has been found to increase with later stages of melanoma. In melanoma cell lines, the over expression of STIM1 enhances invadopodia number whereas STIM1 knockdown inhibits invadopodia formation. Similarly, gelatin degradation activity is enhanced with STIM1 overexpression and abrogated with STIM1 knockdown, implicating STIM1 as an important factor in the regulation of invadopodia formation and melanoma invasion. Though the studies published have shown a significant role of STIM1 in tumor progression, a robust transgenic animal model has not yet been established. Here, we developed a novel transgenic mouse model which, upon 4-hydroxytamoxifen (4OHT) treatment, induces the BRAFV600E mutation and PTEN, STIM1, and STIM2 deletions in melanocytes via an inducible Cre-lox system. Our investigation found that the loss of STIM1 exacerbates tumor growth and results in tumor formation significantly more quickly than STIM1 wild type mice. Whereas PCR analysis of 4OHT-treated skin showed deletion of STIM1 and PTEN, immunohistochemical staining of these genes in tumors did not convincingly demonstrate complete deletion. Therefore, it remains to be determined whether the effects we observed are due to STIM1 and STIM2 loss. These findings need to be corroborated in the future. Our studies focus on two important mechanisms required for melanoma progression and metastasis. We found that α(1,2) fucosylation is able to inhibit invadopodia formation, and melanoma cell invasion. The reestablishment of α(1,2) fucosylation in melanoma could potentially be exploited to inhibit melanoma metastasis. Additionally, early evidence points to STIM1 having a tumor suppressive role in melanoma oncogenesis and tumor growth based on the transgenic mouse model. Although the phenotype is unexpected, further investigation of this model will likely provide important insight for the complicate roles of SOCE in melanoma initiation and progression.

La maladie de l’œdème est une maladie connue depuis de nombreuses années chez le porc. Les premiers cas recensés dans une population de suidés sauvages sont apparus en 2013 en Ardèche. Un nouveau foyer de cette maladie est ensuite apparu en 2016 dans les Pyrénées-Orientales à la frontière entre la France et l’Espagne. Comprendre les facteurs permettant son apparition ainsi que sa transmission est nécessaire afin d’anticiper de futures mortalités dues à cette maladie. Dans cette thèse, une analyse épidémiologique de cette maladie chez le sanglier a été réalisée. Des clusters de mortalités sont alors apparus et ont permis de mettre en évidence une possible source de contamination unique et récurrente dans le temps. La mise en place d’une nouvelle méthode pour étudier la détectabilité des cadavres de sanglier a souligné la difficulté de retrouver des cadavres de sanglier en forêt. La dernière analyse épidémiologique à partir d’un modèle de type « Spatial point pattern » a mis en avant de possibles facteurs de risque d’apparition et de transmission qui ont ensuite été analysés plus précisément. L’analyse des données issus des tableaux de chasse en Ardèche a été réalisée afin de détecter des variations de la densité et du ratio J/A des populations de sanglier suggérant un stress alimentaire chez le sanglier, un prodrome ou une conséquence de la maladie. Aucun stress alimentaire ne fut détecté lors de cette analyse. Des hypothèses ont pu être émises pour expliquer certaines variations observées : i) la conséquence directe de la maladie, ii) un phénomène environnemental particulier et iii) un évènement pathogénique. La piste de l’événement pathogénique a été approfondie avec la découverte du SDRP (syndrome dysgénésique et respiratoire du porc). Les interactions porcs-sangliers, nombreuses en Ardèche, ont été déterminées comme potentiellement responsables du passage de la bactérie entre le compartiment domestique et sauvage. Une étude génétique a également été effectuée pour investiguer le gène alpha-1-fucosyltransferase associé à la sensibilité du porc à la maladie. Tous les sangliers analysés étaient sensibles à la maladie. D’autres analyses complémentaires sont nécessaires afin de comprendre au mieux cette maladie ainsi que les différents facteurs de risque pour l’apparition mais également la transmission. / Edema disease has been a known disease in pigs for many years. The first cases recorded in a population of wild suids appeared in 2013 in Ardèche. A new outbreak of this disease then emerged in 2016 in the Pyrénées-Orientales on the border between France and Spain. Understanding the factors that enable its onset and transmission is necessary to anticipate future mortality from this disease. In this thesis, an epidemiological analysis of this disease in wild boar was carried out. Clusters of mortalities then emerged, highlighting a possible single and recurrent source of contamination over time. The introduction of a new method to study the detectability of wild boar corpses highlighted the difficulty of finding wild boar corpses in the forest. The latest epidemiological analysis using a Spatial point pattern model highlighted possible risk factors for onset and transmission, which were then analysed more precisely. Analysis of data from hunting tables in the Ardèche was carried out in order to detect variations in the density and J/A ratio of wild boar populations suggesting food stress in the wild boar, a prodrome or consequence of the disease. No dietary stress was detected during this analysis. Assumptions could be made to explain some observed variations: i) the direct consequence of the disease, ii) a particular environmental phenomenon and iii) a pathogenic event. The trail of the pathogenic event was deepened with the discovery of the PRRS (Pork Respiratory and Dygesic Syndrome). The pig-boar interactions, numerous in the Ardeche, were determined as potentially responsible for the passage of the bacteria between the domestic and wild compartment. A genetic study was also conducted to investigate the alpha-1-fucosyltransferase gene associated with the susceptibility of pigs to the disease. All the wild boars tested were susceptible to the disease.Further further analysis is needed in order to better understand this disease as well as the different risk factors for both onset and transmission.