Orientador: Flávia Lombardi Lopes / Resumo: O melhoramento genético em bovinos visa a seleção de características para facilitar o manejo, a qualidade da carne, a resistência a doenças e a adaptação ao meio ambiente. Polimorfismos de nucleotídeo único (SNPs) podem gerar grandes efeitos sobre essas características fenotípicas. Os microRNAs são pequenos RNAs não-codificadores que atuam como reguladores da expressão pós-transcricional através de sua ligação a mRNAs alvo. No presente estudo, realizamos o cruzamento de dados entre ~56 milhões de SNPs contra todas as seqüências conhecidas de miRNA bovino e analisamos in silico, seus possíveis efeitos. Seguindo a predição dos alvos, mostramos que 82% dos alvos foram alterados como consequência dos SNPs que ocorrem na região de seed de miRNAs maduros. Em seguida, identificamos variações na Energia Livre Mínima (MFE) que representam a capacidade de alterar a estabilidade das moléculas e, consequentemente, a maturação dos miRNAs. Também encontramos 129 SNPs em miRNAs, que alteraram sua predição com alvos, ocorrendo em regiões de QTL e, por último, a análise dos escores de conservação evolutiva para cada locus de SNP sugeriu que eles têm uma função biológica conservada através do processo evolutivo. Nossos resultados sugerem que os SNPs em microRNAs têm o potencial de alterar os fenótipos bovinos e são de grande valor para a pesquisa de melhoramento genético, bem como para a produção. / Abstract: Genetic improvement of cattle is aimed at selection of characteristics to facilitate the handling, quality of the meat, resistance to diseases and adaptation to the environment. Single nucleotide polymorphisms (SNPs) can generate large effects on these phenotypic characteristics. MicroRNAs are small non-coding RNAs that act as regulators of posttranscriptional expression through their binding to target mRNAs. In the present study, we scanned ~56 million SNPs against all known bovine miRNA sequences and analyzed in silico, their possible effects. Following target prediction, we show that 82% of targets were altered as a consequence of SNPs that occur in the seed region of mature miRNAs. Next, we identified variations in the Minimum Free Energy (MFE) which represent the capacity to alter molecule stability and, consequently, the maturation of the miRNAs. We have also found 129 SNPs in miRNAs, with altered target prediction, occurring in QTL regions and, lastly, analysis of evolutionary conservation scores for each SNP locus suggested that they have a conserved biological function through the evolutionary process. Our results suggest that SNPs in microRNAs have the potential to alter bovine phenotypes and are of great value for genetic improvement research, as well as production. / Mestre

>Magister Scientiae - MSc / Anxiety, resilience and childhood trauma can be categorized as functional behavioural categories, with a wealth of research behind each. The research approach adopted for each, in most cases, is either from a genetic or neuropsychological standpoint, with few studies combining both to better understand all three functional behavioural categories as a multidimensional construct A number of candidate genes have been identified as markers for anxiety, resilience and childhood trauma, of which Catechol-methyl-transferase (COMT) and several respective single nucleotide polymorphisms (SNPs) are included. Although COMT SNPs have been linked to at least one of the functional categories, with a handful of haplotypes identified, to our knowledge no study has investigated the combination of SNPs selected for this study (rs6269, rs4818, rs4680, rs4633, rs737865, rs2075507) as a possible haplotype, specifically in a South African population. The use of SNaPshot for the genotyping of genes is an efficient and reliable means of identifying genotype frequencies and haplotypes in large sample groups, yet when selecting more than two SNPs of interest, the development of a multiplex assay is ideal. The first aim of the study was to design and optimize a multiplex assay to genotype several COMT SNPs. The primer design, multiplex optimization and SNaPshot conditions used showed good working parameters that can be utilized and further improved by optimization. Self-report measures are widely used to measure psychiatric disorders, such as anxiety, and has also been used for the measurement of resilience and childhood trauma. With each functional behavioural category well investigated in its respective domain, there is a need to investigate all three as a collective in a South African population due to the high rate of anxiety and childhood trauma exposure in communities. The second aim of the study was to investigate the prevalence of anxiety, resilience and childhood as functional behavioural categories in the full South African sample group; and the role of sex, through established self-report measures and respective normative data. Additionally, this carried over into investigating the correlation between anxiety, resilience and childhood trauma as a multidimensional construct in both the full South African sample and between sexes. There is a clear relationship which exists between all three functional behavioural categories, as they show a correlation in various dimensions independent of one another. Higher anxiety levels amongst females were reported, with no difference between sexes for resiliency. The empirical data collected from both COMT SNP and self-report measures for male and female where explored and reviewed against current literature for better understanding and insight into the association of COMT SNPs with anxiety, resilience and childhood trauma in a South African population. The results of this study to understand the complexity and association of all three functional behavioural categories as a multidimensional construct will be invaluable and may assist in the identification of possible risk factors which are essential for the promotion of better mental health in society.

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Η πιο απλή μορφή γενετικής διαφοροποίησης στον άνθρωπο είναι οι μονονουκλεοτιδικοί πολυμορφισμοί (Single Nucleotide Polymorphisms - SNPs). Ο αριθμός αυτού του είδους πολυμορφισμών που έχουν βρεθεί στο ανθρώπινο γονιδίωμα και επηρεάζουν την παραγόμενη πρωτεΐνη αυξάνεται συνεχώς, αλλά η αντιστοίχηση τους σε πιθανές ασθένειες με πειραματικές μεθόδους είναι ασύμφορη από θέμα χρόνου και κόστους. Για αυτό τον λόγο έχουν αναπτυχθεί διάφορες υπολογιστικές μέθοδοι με σκοπό να ταξινομήσουν τους μονονουκλεοτιδικούς πολυμορφισμούς σε παθογόνους και μη. Οι περισσότερες από αυτές τις μεθόδους χρησιμοποιούν ταξινομητές, οι οποίοι παίρνοντας σαν είσοδο ένα σύνολο δομικών, λειτουργικών, ακολουθιακών και εξελικτικών χαρακτηριστικών, επιχειρούν να προβλέψουν αν ένας μονονουκλεοτιδικός πολυμορφισμός είναι παθογόνος ή μη. Για την εκπαίδευση αυτών των ταξινομητών, χρησιμοποιούνται δύο σύνολα μονονουκλεοτιδικών πολυμορφισμών. Το πρώτο αποτελείται από μονονουκλεοτιδικούς πολυμορφισμούς που έχει βρεθεί πειραματικά ότι οδηγούν σε παθογένεια και το δεύτερο από μονονουκλεοτιδικούς πολυμορφισμούς που έχει αποδειχθεί πειραματικά ότι είναι αδρανείς. Οι μέθοδοι αυτές διαφέρουν στα χαρακτηριστικά των μεταλλάξεων που λαμβάνουν υπόψη στην πρόβλεψη τους, καθώς επίσης και στην εκπαίδευση και τη φύση των τεχνικών ταξινόμησης, που χρησιμοποιούν για τη λήψη των αποφάσεων. Το βασικότερο προβλήματα τους ωστόσο έγκειται στο γεγονός ότι καθορίζουν τα χαρακτηριστικά, που θα χρησιμοποιήσουν σαν είσοδο στους ταξινομητές τους με τρόπο εμπειρικό και μάλιστα διαφορετικές μέθοδοι προτείνουν και χρησιμοποιούν διαφορετικά χαρακτηριστικά, χωρίς να τεκμηριώνουν επαρκώς τις αιτίες αυτής της διαφοροποίησης. Δύο ακόμα προβλήματα που δεν έχουν καταφέρει να αντιμετωπίσουν οι υπάρχουσες μεθοδολογίες είναι το πρόβλημα της ανισορροπίας των δύο κλάσεων ταξινόμησης και των ελλιπών τιμών σε πολλά από τα χαρακτηριστικά εισόδου των ταξινομητών, ώστε να επιτυγχάνουν πιο ακριβή και αξιόπιστα αποτελέσματα. Από τα παραπάνω είναι ξεκάθαρο πως υπάρχει μεγάλο περιθώριο βελτίωσης των υπάρχουσων μεθοδολογιών για το συγκεκριμένο πρόβλημα ταξινόμησης. Στην παρούσα διπλωματική εργασία προτείνουμε μια νέα υβριδική μεθοδολογία υπολογιστικής νοημοσύνης, που ξεπερνά πολλά από τα προβλήματα των υπάρχοντων μεθοδολογιών και βελτιώνει με τον τρόπο αυτό την απόδοσή τους. Δύο είναι τα βασικά βήματα που ακολουθήσαμε για την επίτευξη του στόχου αυτού. Πρώτον, συγκεντρώσαμε από τις διαθέσιμες δημόσιες βάσεις δεδομένων, τους μονονουκλεοτιδικούς πολυμορφισμούς που χρησιμοποιήθηκαν για την εκπαίδευση και τον έλεγχο των μοντέλων μηχανικής μάθησης. Συγκεκριμένα, συλλέχθησαν και φιλτραρίστηκαν τα θετικά και αρνητικά σύνολα εκπαίδευσης και ελέγχου, που αποτελούνται από μονονουκλεοτιδικούς πολυμορφισμούς που είτε οδηγούν σε παθογένεια, είτε είναι ουδέτεροι. Για κάθε πολυμορφισμό των δύο συνόλων υπολογίσαμε χρησιμοποιώντας υπάρχοντα διαθέσιμα εργαλεία όσο το δυνατό περισσότερα δομικά, λειτουργικά, ακολουθιακά και εξελικτικά χαρακτηριστικά. Για εκείνα τα χαρακτηριστικά, για τα οποία δεν υπήρχε κάποιο διαθέσιμο εργαλείο υπολογισμού τους, υλοποιήσαμε τον κατάλληλο κώδικα για τον υπολογισμό τους. Το δεύτερο βήμα της διπλωματικής αφορούσε το σχεδιασμό και την υλοποίηση της κατάλληλης υβριδικής μεθόδου για την επίλυση του προβλήματος που μελετάμε. Χρησιμοποιήσαμε μια νέα μέθοδο ταξινόμησης την EnsembleGASVR. Πρόκειται για μια ensemble μεθοδολογία, που συνδυάζει σε ένα ενιαίο πλαίσιο ταξινόμησης οκτώ διαφορετικούς ταξινομητές. Κάθε ένας από αυτούς τους ταξινομητές βασίζεται στον υβριδικό συνδυασμό των Γενετικών Αλγορίθμων και των μοντέλων Παλινδρόμησης Διανυσμάτων Υποστήριξης (nu-Support Vector Regression). Συγκεκριμένα ένας Προσαρμοζόμενος Γενετικός Αλγόριθμος χρησιμοποιείται για να καθοριστεί το βέλτιστο υποσύνολο χαρακτηριστικών, καθώς και οι βέλτιστες τιμές των παραμέτρων των ταξινομητών. Σαν μέθοδο ταξινόμησης των μεταλλάξεων σε ουδέτερες και παθογενείς, προτείνουμε τον nu-SVR ταξινομητή, καθώς παρουσιάζει υψηλή απόδοση, καλή γενίκευση, δεν παγιδεύεται σε τοπικά βέλτιστα, ενώ ταυτόχρονα επιτυγχάνει την ισορροπία μεταξύ της ακρίβειας και της πολυπλοκότητας του μοντέλου. Μάλιστα για να ξεπεράσουμε τα πρόβληματα των ελλιπών τιμών και της ανισορροπίας των δύο κλάσεων ταξινόμησης, αλλά και για να βελτιώσουμε τη συνολική απόδοση της μεθοδολογίας μας, επεκτείναμε τον υβριδικό αλγόριθμο, ώστε να λειτουργεί σαν μία ensemble-συλλογική τεχνική, συνδυάζοντας οκτώ επί μέρους μοντέλα ταξινόμησης. Τα πειραματικά αποτελέσματα της προτεινόμενης μεθοδολογίας ήταν εξαιρετικά ελπιδοφόρα, καθώς η EnsembleGASVR μεθοδολογία υπερτερεί σημαντικά έναντι άλλων ευρέως γνωστών μεθόδων ταξινόμησης παθογενών μεταλλάξεων. / Single Nucleotide Polymorphisms (SNPs) are the most common form of genetic variations in humans. The number of SNPs that have been found in human genome and affect protein functionality is constantly increasing. Finding matches between SNPs and diseases using experimental techniques, is excessive disadvantageous in terms of time and cost. For this reason, several computational methods have been developed. These methods classify polymorphisms as pathogenic and non-pathogenic. Most of them use classifiers, which take as input a set of structural, functional, sequential and evolutionary features and predict whether a single nucleotide polymorphism is pathogenic or neutral. For training these classifiers use two sets of SNPs. The first one consists of SNPs that have been experimentally proven as pathogenic, whereas the second set consists of SNPs that have been experimentally characterized as benign. These methods differ in the classification methods they deploy and in the features they use as inputs. However, the main problem is the determination of an empirically verified set of features for training. Specifically, different methods suggest different feature sets, without adequately documenting the causes of this differentiation. In addition, the existing methodologies do not tackle efficiently the class imbalance problem between positive and negative training sets and the problem of missing values in the datasets. In this thesis a new hybrid computational intelligence methodology is proposed, that overcomes many of the problems of existing methodologies. The proposed method achieves high classification performance and systematizes the selection of relevant features. In the first phase of this study the polymorphisms were gathered from the available public databases and they were used for training and testing of the machine learning models. Specifically, the positive and negative training and test sets were collected and filtered. They consist of single nucleotide polymorphisms that lead to either pathogenesis or are neutral. For each polymorphism of the two sets, using existing available tools, a wide range of structural, functional, sequential and evolutionary features were calculated. For those features for which there was no available tool, the suitable program (code) was developed in order to compute them. In the second step a new embedded hybrid classification method called EnsembleGASVR is designed and implemented. The method uses an ensemble methodology, based on hybrid combination of Genetic Algorithms and nu-Support Vector Regression (nu-SVR) models. An Adaptive Genetic Algorithm is used to determine the optimal subset of features and the optimal values of the parameters of classifiers. We propose the nu-SVR classifier, since it exhibits high performance, good generalization ability, it is not trapped in local optima and achieves a balance between accuracy and complexity of the model. In order to overcome the problem of missing values and class imbalance, we extended the above algorithm to function as a collective ensemble-technique, combining eight individual classification models. In overall, the method achieves 87.45% accuracy, 71.78% sensitivity and 93.16% specificity. These priliminary results are very promising and shows that EnsembleGASVR methodology significantly outperforms other well-known classification methods for pathogenic mutations.

Female reproductive performance is a critical component of sustainable pig production systems. There is abundant evidence of genetic variation in these traits among pig breeds. The aims of this study were to identify quantitative trait loci (QTL) affecting reproductive traits and to identify and characterise positional candidate gene(s) underlying the QTL. A Large White - Meishan F2 population was scanned for QTL with effects on reproductive traits. This analysis revealed 13 putative QTLs on seven different chromosomes with effects on five different traits: ovulation rate (OR), teat number (TN), prenatal survival (PS), total born alive (TBA) and litter size (LS). QTL for PS and LS on chromosome 8 were fine mapped and Secreted Phosphoprotein 1 (SPP1) confirmed as a candidate gene. A genome-wide association study was performed on a diverse population of different breeds and crosses lines, for reproductive traits including LS, TBA, number of stillborn piglets, and number of mummified piglets. Fourteen SNPs were found significantly associated with reproductive traits. The functional study of SPP1 examined the hypothesis that differences in foetal growth may be associated with the effectiveness of conceptus attachment, as measured by SPP1 expression. Patterns of SPP1 mRNA and protein expression in placental and uterine tissues supplying the smallest and a normal-sized foetus from the same uterus were examined in Large White-Landrace (LW-LR), Large White (LW) and Meishan (MS) females 40 and 45 of pregnancy. The smallest LW-LR foetuses tended to have a higher level of SPP1 mRNA in endometrium tissue compared to the normal-sized foetuses. However, placenta expression was higher in the normal-sized foetuses compared to the smallest ones. SPP1 protein levels in normal sized foetuses were significantly higher than in the smallest litter mates for all the tissues. Significantly higher levels of SPP1 mRNA and protein were found in MS compared to LW. In both breeds, significant differences between sizes were found in some tissues, with similar expression patterns in respect to size, for both mRNA and protein in endometrial tissues when compared to contemporary LW. In placenta, the direction of the expression differed between breeds, with a higher expression of mRNA and protein in the normal-sized MS foetuses and in the smallest sized LW foetuses. The comparison of SPP1 expression between different foetal sizes and different breeds revealed associations between breed, foetal size, and SPP1 protein, factors implicated in PS and LS. These results together with the genetic evidence indicate that the potential role of SPP1 in placental and foetal development merits further investigation.

Depuis le premier séquençage du génome humain au début des années 2000, de grandes initiatives se sont lancé le défi de construire la carte des variabilités génétiques inter-individuelles, ou bien encore celle des altérations de l'ADN tumoral. Ces projets ont posé les fondations nécessaires à l'émergence de la médecine de précision, dont le but est d'intégrer aux dossiers médicaux conventionnels les spécificités génétiques d'un individu, afin de mieux adapter les traitements et les stratégies de prévention. La traduction des variations et des altérations de l'ADN en prédictions phénotypiques constitue toutefois un problème difficile. Les séquenceurs ou puces à ADN mesurent plus de variables qu'il n'y a d'échantillons, posant ainsi des problèmes statistiques. Les données brutes sont aussi sujettes aux biais techniques et au bruit inhérent à ces technologies. Enfin, les vastes réseaux d'interactions à l'échelle des protéines obscurcissent l'impact des variations génétiques sur le comportement de la cellule, et incitent au développement de modèles prédictifs capables de capturer un certain degré de complexité.Cette thèse présente de nouvelles contributions méthodologiques pour répondre à ces défis.Tout d'abord, nous définissons une nouvelle représentation des profils de mutations tumorales, qui exploite leur position dans les réseaux d'interaction protéine-protéine. Pour certains cancers, cette représentation permet d'améliorer les prédictions de survie à partir des données de mutations, et de stratifier les cohortes de patients en sous-groupes informatifs. Nous présentons ensuite une nouvelle méthode d'apprentissage permettant de gérer conjointement la normalisation des données et l'estimation d'un modèle linéaire. Nos expériences montrent que cette méthode améliore les performances prédictives par rapport à une gestion séquentielle de la normalisation puis de l'estimation. Pour finir, nous accélérons l'estimation de modèles linéaires parcimonieux, prenant en compte des interactions deux à deux, grâce à un nouvel algorithme. L'accélération obtenue rend cette estimation possible et efficace sur des jeux de données comportant plusieurs centaines de milliers de variables originales, permettant ainsi d'étendre la portée de ces modèles aux données des études d'associations pangénomiques. / Since the first sequencing of the human genome in the early 2000s, large endeavours have set out to map the genetic variability among individuals, or DNA alterations in cancer cells. They have laid foundations for the emergence of precision medicine, which aims at integrating the genetic specificities of an individual with its conventional medical record to adapt treatment, or prevention strategies.Translating DNA variations and alterations into phenotypic predictions is however a difficult problem. DNA sequencers and microarrays measure more variables than there are samples, which poses statistical issues. The data is also subject to technical biases and noise inherent in these technologies. Finally, the vast and intricate networks of interactions among proteins obscure the impact of DNA variations on the cell behaviour, prompting the need for predictive models that are able to capture a certain degree of complexity. This thesis presents novel methodological contributions to address these challenges. First, we define a novel representation for tumour mutation profiles that exploits prior knowledge on protein-protein interaction networks. For certain cancers, this representation allows improving survival predictions from mutation data as well as stratifying patients into meaningful subgroups. Second, we present a new learning framework to jointly handle data normalisation with the estimation of a linear model. Our experiments show that it improves prediction performances compared to handling these tasks sequentially. Finally, we propose a new algorithm to scale up sparse linear models estimation with two-way interactions. The obtained speed-up makes this estimation possible and efficient for datasets with hundreds of thousands of main effects, thereby extending the scope of such models to the data from genome-wide association studies.

>Magister Scientiae - MSc / As human beings we are curious about our origin and ancestry. A curiosity has led to an investigation of human evolution and expansion across the world by means of population genetics and phylo-genetics by evaluating a region in Southern Africa that is largely unknown. The objective of this study was to develop a quick, inexpensive and accurate hierarchical diagnostic screening system of the MtDNA phylogenetic tree, AI-SNPs in the mtDNA genome by using High Resolution Melting analysis to evaluate the population composition and ancestral haplogroups of Southern African populations in Limpopo. The admixture between the ‘Khoesan’ hunter-gatherers, herders and the Bantu speaking populations led to population growth and expansion in Limpopo. This has contributed to populations settling in Limpopo and has thus shaped the ancestral contemporary populations. No research on these individuals residing in Limpopo has been done before, thus an investigation of their ancestral origin was necessary. A total of 760 saliva samples were collected from individuals residing in Limpopo. Only 500 saliva samples were extracted by means of an optimized salting out technique. Five hundred extracted genomic samples were genotyped by means of a quick, inexpensive High-resolution melting analysis. Of the 500 samples, the genotyping results showed 95 individuals derived for the L3 haplogroup which gives a 19% ratio of individuals screened with Multiplex 1. Only 56 individuals were derived for the L1 haplogroup, which gives a percentage of 11%. A total of 249 individuals were derived for the L0 haplogroup, making up a 50% of the total individuals genotyped. Only 100 samples were derived for L0a, making up 20% of individuals screened with Multiplex 1. Of the 95 samples derived for the L3 haplogroup, the results showed 87 individuals to be ancestral for both M and N, making up 91.57% of individuals screened with Multiplex 2. http://etd.uwc.ac.za/. In population genetics using SNPs to infer population history and ancestral origin has become significant, this study allowed researchers to evaluate population groups by investigating their genetic markers and the application of the results allowed for downstream analyses. Finally, this study provides a quick and simple screening method for the selection of lineages that are of interest for further studies.

Contexte - La variation interindividuelle de la réponse aux corticostéroïdes (CS) est un problème important chez les patients atteints de maladies inflammatoires d’intestin. Ce problème est bien plus accentué chez les enfants avec la prévalence de la corticodépendance extrêmement (~40 %) élevée. La maladie réfractaire au CS a des répercussions sur le développement et le bien-être physique et psychologique des patients et impose des coûts médicaux élevés, particulièrement avec la maladie active comparativement à la maladie en rémission, le coût étant 2-3 fois plus élevé en ambulatoire et 20 fois plus élevé en hôpital. Il est ainsi primordial de déterminer les marqueurs prédictifs de la réponse aux CS. Les efforts précédents de découvrir les marqueurs cliniques et démographiques ont été équivoques, ce qui souligne davantage le besoin de marqueurs moléculaires. L'action des CS se base sur des processus complexes déterminés génétiquement. Deux gènes, le ABCB1, appartenant à la famille des transporteurs transmembraneaux, et le NR3C1, encodant le récepteur glucocorticoïde, sont des éléments importants des voies métaboliques. Nous avons postulé que les variations dans ces gènes ont un rôle dans la variabilité observée de la réponse aux CS et pourraient servir en tant que les marqueurs prédictifs. Objectifs - Nous avons visé à: (1) examiner le fardeau de la maladie réfractaire aux CS chez les enfants avec la maladie de Crohn (MC) et le rôle des caractéristiques cliniques et démographiques potentiellement liés à la réponse; (2) étudier l'association entre les variantes d'ADN de gène ABCB1 et la réponse aux CS; (3) étudier les associations entre les variantes d'ADN de gène NR3C1 et la réponse aux CS. Méthodes - Afin d’atteindre ces objectifs, nous avons mené une étude de cohorte des patients recrutés dans deux cliniques pédiatriques tertiaires de gastroentérologie à l’Ottawa (CHEO) et à Montréal (HSJ). Les patients avec la MC ont été diagnostiqués avant l'âge de 18 ans selon les critères standard radiologiques, endoscopiques et histopathologiques. La corticorésistance et la corticodépendance ont été définies en adaptant les critères reconnus. L’ADN, acquise soit du sang ou de la salive, était génotypée pour des variations à travers de gènes ABCB1 et NR3C1 sélectionnées à l’aide de la méthodologie de tag-SNP. La fréquence de la corticorésistance et la corticodépendance a été estimée assumant une distribution binomiale. Les associations entre les variables cliniques/démographiques et la réponse aux CS ont été examinées en utilisant la régression logistique en ajustant pour des variables potentielles de confusion. Les associations entre variantes génétiques de ABCB1 et NR3C1 et la réponse aux CS ont été examinées en utilisant la régression logistique assumant différents modèles de la transmission. Les associations multimarqueurs ont été examinées en utilisant l'analyse de haplotypes. Les variantes nongénotypées ont été imputées en utilisant les données de HAPMAP et les associations avec SNPs imputés ont été examinées en utilisant des méthodes standard. Résultats - Parmi 645 patients avec la MC, 364 (56.2%) ont reçu CS. La majorité de patients étaient des hommes (54.9 %); présentaient la maladie de l’iléocôlon (51.7%) ou la maladie inflammatoire (84.6%) au diagnostic et étaient les Caucasiens (95.6 %). Huit pourcents de patients étaient corticorésistants et 40.9% - corticodépendants. Le plus bas âge au diagnostic (OR=1.34, 95% CI: 1.03-3.01, p=0.040), la maladie cœxistante de la région digestive supérieure (OR=1.35, 95% CI: 95% CI: 1.06-3.07, p=0.031) et l’usage simultané des immunomodulateurs (OR=0.35, 95% CI: 0.16-0.75, p=0.007) ont été associés avec la corticodépendance. Un total de 27 marqueurs génotypés à travers de ABCB1 (n=14) et NR3C1 (n=13) ont été en l'Équilibre de Hardy-Weinberg, à l’exception d’un dans le gène NR3C1 (rs258751, exclu). Dans ABCB1, l'allèle rare de rs2032583 (OR=0.56, 95% CI: 0.34-0.95, p=0.029) et génotype hétérozygote (OR=0.52, 95% CI: 0.28-0.95 p=0.035) ont été négativement associes avec la dépendance de CS. Un haplotype à 3 marqueurs, comprenant le SNP fonctionnel rs1045642 a été associé avec la dépendance de CS (p empirique=0.004). 24 SNPs imputés introniques et six haplotypes ont été significativement associés avec la dépendance de CS. Aucune de ces associations n'a cependant maintenu la signification après des corrections pour des comparaisons multiples. Dans NR3C1, trois SNPs: rs10482682 (OR=1.43, 95% CI: 0.99-2.08, p=0.047), rs6196 (OR=0.55, 95% CI: 0.31-0.95, p=0.024), et rs2963155 (OR=0.64, 95% CI: 0.42-0.98, p=0.039), ont été associés sous un modèle additif, tandis que rs4912911 (OR=0.37, 95% CI: 0.13-1.00, p=0.03) et rs2963156 (OR=0.32, 95% CI: 0.07-1.12, p=0.047) - sous un modèle récessif. Deux haplotypes incluant ces 5 SNPs (AAACA et GGGCG) ont été significativement (p=0.006 et 0.01 empiriques) associés avec la corticodépendance. 19 SNPs imputés ont été associés avec la dépendance de CS. Deux haplotypes multimarqueurs (p=0.001), incluant les SNPs génotypés et imputés, ont été associés avec la dépendance de CS. Conclusion - Nos études suggèrent que le fardeau de la corticodépendance est élevé parmi les enfants avec le CD. Les enfants plus jeunes au diagnostic et ceux avec la maladie coexistante de la région supérieure ainsi que ceux avec des variations dans les gènes ABCB1 et NR3C1 étaient plus susceptibles de devenir corticodépendants. / Background - Inter-individual variation in response to treatment by corticosteroids (CS) is an important problem in the management of inflammatory bowel disease (IBD) patient’s. This problem is even more prominent in children, the prevalence of steroid dependence (~40%) in whom is extremely high. Steroid refractoriness has a considerable impact on the physical and psychological development of these children, also imposing high medical costs related to treatment. Active disease, as opposed to quiescent, increases medical costs 2-3 times in ambulatory patients and 20 times in hospitalized cases. Identifying markers that could predict steroid response is therefore a high clinical priority. Previous attempts to investigate potential clinical and demographic markers have been equivocal, highlighting the need for further investigations of other predictive markers. It is well known that the action of CS entails complex processes controlled by genetic factors. Two genes, the ABCB1 gene, which belongs to the family of trans-membrane transporters, and the NR3C1 gene, coding for the glucocorticoid receptor, are major elements of the pathway. We postulated that inter-individual variations in these genes may play a role in the observed variability of the response to CS and could serve as potential predictors. Objectives - We aimed to: (1) examine the burden of steroid refractoriness in children diagnosed with CD and explore the potential clinical/demographic factors related to CS response; (2) study the association between DNA variants in the ABCB1 gene and CS response; (3) investigate the associations between DNA variants in the NR3C1 gene and CS response. Methods - We investigated these objectives in a cohort of CD patients recruited from two tertiary paediatric gastroenterology clinics from Ottawa (CHEO) and Montreal (HSJ). CD patients diagnosed prior to age 18 using standard clinical, radiological, endoscopic and histopathological criteria were included. Published criteria were adapted to define CS-resistance and dependence. DNA acquired from blood and/or saliva was genotyped for variations across the ABCB1 and NR3C1 genes selected using the tag-SNP methodology. The frequencies of steroid resistance and dependence were estimated assuming a binomial distribution. Associations between clinical/demographic variables and steroid responses were examined using logistic regression modeling after accounting for potential confounding variables. Associations between ABCB1 and NR3C1 genes’ variants and steroid responses were examined using logistic regression assuming different models of inheritance. Multi-marker associations were examined via haplotype analysis. Un-genotyped variants in the genes were imputed using HAPMAP data as the reference panel and associations with imputed SNPs examined using standard methods. Results - Among 645 CD patients diagnosed at the study centers, 364 (56.2%) received corticosteroids during the first year since diagnosis. The majority of patients were male (54.9%), had inflammatory (84.6%), ileo-colonic (51.7%) disease phenotypes at diagnosis and were Caucasians (95.6%). Eight percent of patients developed CS-resistance and 40.9% became CS-dependent. Younger age at diagnosis (OR=1.34, 95% CI: 1.03-3.01, p=0.040), coexisting upper digestive tract involvement (OR=1.35, 95% CI: 1.06-3.07, p=0.031) and concomitant immunomodulators use (OR=0.35, 95% CI: 0.16-0.75, p=0.007) were significantly associated with CS-dependency in multivariate analysis. From among the 27 markers genotyped across the ABCB1 (n=14) and NR3C1 genes (n=13), all except one in NR3C1 gene (rs258751, excluded) were in Hardy-Weinberg Equilibrium. For ABCB1, the rare allele of rs2032583 (OR=0.56, 95% CI: 0.34-0.95, p=0.029) and heterozygous genotype (OR=0.52, 95% CI: 0.28-0.95, p=0.035) conferred protection from CS dependency. A 3-marker haplotype including the functional SNP rs1045642 was associated with CS-dependence (empiric p-value=0.004). On imputation 24 intronic SNPs and six haplotypes were statistically significantly associated with CS dependence. None of these associations however maintained significance after corrections for multiple comparisons. For the NR3C1 gene 3 SNPs, rs10482682 (OR=1.43, 95% CI: 0.99-2.08, p=0.047), rs6196 (OR=0.55, 95% CI: 0.31-0.95, p=0.024), and rs2963155 (OR=0.64, 95% CI: 0.42-0.98, p=0.039), showed associations under an additive model whereas rs4912911 (OR=0.37, 95% CI: 0.13-1.00, p=0.03) and rs2963156 (OR=0.32, 95% CI: 0.07-1.12, p=0.047) showed associations under a recessive model. Two haplotypes encompassing these 5 SNPs (AAACA and GGGCG) were significantly (empirical p=0.006 and 0.01 respectively) were associated with CS-dependence. On imputation 19 SNPs were associated with CS-dependence. Two multi-marker haplotypes (p-values=0.001 each) including genotyped and imputed SNPs conferred susceptibility for CS-dependency. Conclusions - Our studies suggest that the burden of steroid dependence is high among children with CD. Children diagnosed at a younger age, those with co-existent upper tract disease and with variations in the ABCB1 and NR3C1 genes were more likely to become CS dependent.

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El treball presentat en aquesta tesi és un estudi de la diversitat genètica en un conjunt de gens implicats en funcions neurològiques ("Gens cerebrals"). Hom ha examinat vint-i-dos gens implicats en els sistemes de neurotransmissió dopaminèrgic, serotoninèrgic i glutamatèrgic. L'objectiu de l'estudi té dos vessants: per una banda l'anàlisi de la diversitat genètica en un conjunt de gens implicats en malaltia humana, en aquest cas en malaltia psiquiàtrica, en poblacions humanes mundials, amb la intenció d'assentar les bases per propers esforços de mapatge genètic; i per altra banda analitzar la diversitat genètica en aquest conjunt de gens per tal de descobrir evidències d'esdeveniments històrics, incloent possibles evidències de selecció. / The work presented in this thesis is a study of the genetic variation in a set of genes related to neurological function ('Brain genes'). Twenty two genes are examined, all of which are involved in either the Dopaminergic, Serotonergic or the Glutamatergic systems of neurotransmission.The objective of the study has two aspects: on the one hand the analysis of genetic variation in a set of genes which are implicated in human disease, in this case psychiatric disease, across global human populations, towards the end of providing some new insight for gene mapping efforts, and on the other hand the study of genetic variation in this set of genes may reveal traces of the population history events undergone, including possible evidence for selection.