Dissertations / Theses on the topic 'Metabolomic signature'

Doutoramento em Química
This thesis reports the application of metabolomics to human tissues and biofluids (blood plasma and urine) to unveil the metabolic signature of primary lung cancer. In Chapter 1, a brief introduction on lung cancer epidemiology and pathogenesis, together with a review of the main metabolic dysregulations known to be associated with cancer, is presented. The metabolomics approach is also described, addressing the analytical and statistical methods employed, as well as the current state of the art on its application to clinical lung cancer studies. Chapter 2 provides the experimental details of this work, in regard to the subjects enrolled, sample collection and analysis, and data processing. In Chapter 3, the metabolic characterization of intact lung tissues (from 56 patients) by proton High Resolution Magic Angle Spinning (HRMAS) Nuclear Magnetic Resonance (NMR) spectroscopy is described. After careful assessment of acquisition conditions and thorough spectral assignment (over 50 metabolites identified), the metabolic profiles of tumour and adjacent control tissues were compared through multivariate analysis. The two tissue classes could be discriminated with 97% accuracy, with 13 metabolites significantly accounting for this discrimination: glucose and acetate (depleted in tumours), together with lactate, alanine, glutamate, GSH, taurine, creatine, phosphocholine, glycerophosphocholine, phosphoethanolamine, uracil nucleotides and peptides (increased in tumours). Some of these variations corroborated typical features of cancer metabolism (e.g., upregulated glycolysis and glutaminolysis), while others suggested less known pathways (e.g., antioxidant protection, protein degradation) to play important roles. Another major and novel finding described in this chapter was the dependence of this metabolic signature on tumour histological subtype. While main alterations in adenocarcinomas (AdC) related to phospholipid and protein metabolisms, squamous cell carcinomas (SqCC) were found to have stronger glycolytic and glutaminolytic profiles, making it possible to build a valid classification model to discriminate these two subtypes. Chapter 4 reports the NMR metabolomic study of blood plasma from over 100 patients and near 100 healthy controls, the multivariate model built having afforded a classification rate of 87%. The two groups were found to differ significantly in the levels of lactate, pyruvate, acetoacetate, LDL+VLDL lipoproteins and glycoproteins (increased in patients), together with glutamine, histidine, valine, methanol, HDL lipoproteins and two unassigned compounds (decreased in patients). Interestingly, these variations were detected from initial disease stages and the magnitude of some of them depended on the histological type, although not allowing AdC vs. SqCC discrimination. Moreover, it is shown in this chapter that age mismatch between control and cancer groups could not be ruled out as a possible confounding factor, and exploratory external validation afforded a classification rate of 85%. The NMR profiling of urine from lung cancer patients and healthy controls is presented in Chapter 5. Compared to plasma, the classification model built with urinary profiles resulted in a superior classification rate (97%). After careful assessment of possible bias from gender, age and smoking habits, a set of 19 metabolites was proposed to be cancer-related (out of which 3 were unknowns and 6 were partially identified as N-acetylated metabolites). As for plasma, these variations were detected regardless of disease stage and showed some dependency on histological subtype, the AdC vs. SqCC model built showing modest predictive power. In addition, preliminary external validation of the urine-based classification model afforded 100% sensitivity and 90% specificity, which are exciting results in terms of potential for future clinical application. Chapter 6 describes the analysis of urine from a subset of patients by a different profiling technique, namely, Ultra-Performance Liquid Chromatography coupled to Mass Spectrometry (UPLC-MS). Although the identification of discriminant metabolites was very limited, multivariate models showed high classification rate and predictive power, thus reinforcing the value of urine in the context of lung cancer diagnosis. Finally, the main conclusions of this thesis are presented in Chapter 7, highlighting the potential of integrated metabolomics of tissues and biofluids to improve current understanding of lung cancer altered metabolism and to reveal new marker profiles with diagnostic value.
A presente tese reporta a aplicação da metabolómica ao estudo de tecidos e biofluidos humanos (plasma sanguíneo e urina), com o intuito de caracterizar a assinatura metabólica do cancro pulmonar primário. No Capítulo 1, apresenta-se uma breve introdução sobre a epidemiologia e a patogénese deste tipo de cancro, bem como um sumário das principais alterações metabólicas tipicamente associadas ao cancro em geral. Descreve-se ainda a abordagem metabolómica, nomeadamente os métodos analíticos e estatísticos utilizados, assim como o estado da arte da sua aplicação em estudos clínicos do cancro do pulmão. No Capítulo 2, apresentam-se os detalhes experimentais deste trabalho, no que diz respeito ao grupo de indivíduos envolvidos, à colheita e análise das amostras e ao posterior tratamento dos dados. O Capítulo 3 descreve a caracterização metabólica de tecidos do pulmão (de 56 doentes) por espetroscopia de Ressonância Magnética Nuclear (RMN) de alta resolução com rotação no ângulo mágico. Após a otimização cuidada das condições de aquisição e a identificação detalhada dos sinais espetrais (mais de 50 metabolitos identificados), os perfis metabólicos dos tumores e dos tecidos adjacentes não envolvidos (controlos) foram comparados por análise multivariada, tendo sido discriminados com uma exatidão de 97%. Os metabolitos que mais significativamente contribuíram para esta diferenciação foram: glucose e acetato (diminuídos nos tumores), lactato, alanina, glutamato, GSH, taurina, creatina, fosfocolina, glicerofosfocolina, fosfoetanolamina, nucleótidos de uracilo e péptidos (aumentados nos tumores). Algumas destas variações corroboraram alterações típicas do metabolismo do cancro (e.g., glicólise e glutaminólise aumentadas), enquanto outras sugeriram novas pistas sobre a possível relevância de processos como a proteção antioxidante e a degradação proteica. Um outro resultado novo e importante descrito neste capítulo foi a dependência da assinatura metabólica em relação ao tipo histológico do tumor. Enquanto as principais alterações observadas nos adenocarcinomas (AdC) se relacionaram com o metabolismo fosfolipídico e proteico, os carcinomas de células escamosas (SqCC) apresentaram perfis glicolíticos e glutaminolíticos mais pronunciados, sendo possível construir um modelo válido para a discriminação destes subtipos. No Capítulo 4, apresenta-se o estudo metabolómico por RMN de plasma sanguíneo de mais de 100 doentes e quase 100 controlos saudáveis, do qual resultou um modelo multivariado com uma taxa de classificação de 87%. A distinção entre os grupos foi feita essencialmente com base nos níveis de lactato, piruvato, acetoacetato, lipoproteínas LDL+VLDL e glicoproteínas (aumentados nos doentes), juntamente com os níveis de glutamina, histidina, valina, metanol, lipoproteínas HDL e dois compostos não identificados (diminuídos nos doentes). Estas variações foram detetadas desde os estádios iniciais da doença e a magnitude de algumas delas dependeu do tipo histológico, embora não permitindo discriminar AdC de SqCC. Para além disso, mostra-se neste capítulo que o desequilíbrio dos grupos controlo e cancro em termos da idade dos indivíduos poderá ter alguma influência nos resultados, e apresenta-se uma tentativa exploratória de validação externa, que resultou numa taxa de classificação de 85%. O estudo por RMN do perfil metabólico da urina dos doentes com cancro do pulmão e dos controlos é apresentado no Capítulo 5. Comparativamente ao plasma, o modelo construído com os perfis urinários apresentou uma taxa de classificação superior (97%). Após uma avaliação cuidada da possível influência do género, idade e hábitos tabágicos, um conjunto de 19 metabolitos foi proposto como estando relacionado com a doença (incluindo 3 compostos desconhecidos e 6 parcialmente identificados como metabolitos N-acetilados). Tal como no caso do plasma, estas variações foram detetadas em doentes no estádio inicial e mostraram alguma dependência em relação ao tipo histológico, obtendo-se um modelo válido para a discriminação AdC vs. SqCC, ainda que com um poder preditivo modesto. Para além disso, o teste preliminar de validação externa revelou 100% de sensibilidade e 90% de especificidade, o que é um resultado bastante promissor em termos da potencial utilização dos perfis urinários em aplicações clínicas futuras. No Capitulo 6, descreve-se a caracterização dos perfis metabólicos da urina (de um subgrupo de indivíduos) por cromatografia líquida de ultra-eficiência acoplada a espetrometria de massa (UPLC-MS). Embora não avançando muito na identificação estrutural de possíveis marcadores, este estudo reforçou o valor diagnóstico da urina, já que os modelos multivariados resultantes apresentaram taxa de classificação e poder preditivo elevados. Finalmente, no Capítulo 7, apresentam-se as principais conclusões deste trabalho, realçando o contributo da metabolómica integrada de tecidos e biofluidos para a compreensão do metabolismo alterado do cancro do pulmão e para a deteção de novos perfis marcadores com valor diagnóstico.

Nous avons réalisé une approche métabolomique ciblée par spectrométrie de masse, avec quantification de 188 métabolites incluant des lipides et des molécules polaires. Trois processus physiopathologiques en rapport avec la rétine et le nerf optique ont été étudiés : la neuropathie optique héréditaire de Leber (NOHL), l’atrophie optique autosomique dominante (AOD) par haplo-insuffisance du gène OPA1 et le préconditionnement rétinien induit par la lumière (PRIL). Les principaux résultats sont : Projet NOHL : Le pool des acides aminés et la concentration de certaines sphingomyélines (SM) sont diminués tandis que la concentration de 10 phosphatidylcholines (PC) est augmentée dans les fibroblastes des porteurs d’une mutation de type NOH. Les fibroblastes des patients atteints d’une NOHL ont montré un stress du réticulum endoplasmique réversible pharmacologiquement. Projet AOD : Les variations dans la concentration de certains lipides, du glutamate et de quelques métabolites polaires neuroprotecteurs ont suggéré une altération de la myéline et une dysfonction métabolique axonale pré-symptomatique dans le nerf optique des souris Opa1+/-. Un dimorphisme sexuel a été observé dans le métabolome du nerf optique des souris. Projet PRIL : Le stimulus lumineux préconditionnant semble provoquer un accroissement de la protéolyse et une diminution du monoxyde d’azote dans la rétine. Le stress photique parait associé à un remodelage des lipides rétiniens. Un dimorphisme sexuel a été observé dans le métabolome rétinien des rats contrôles. Ces résultats montrent que l'approche métabolomique est pertinente pour l'étude physiopathologique des maladies oculaires
We have conducted a mass spectrometry targeted metabolomics approach, enabling us to quantify 188 metabolites including lipids and more polar molecules. Three condition related to the retina and the optic nerve have been studied: Leber hereditary optic neuropathy (LHON), dominant optic atrophy (DOA) due to OPA1 haploinsufficiency and retina light-induced preconditioning (RLIP). The main results we obtained are: LHON project: Concentrations of the whole pool of amino acid and some sphingomyelins (SM) were diminished whereas those of ten phosphatidylcholines (PC)were increased in fibroblasts carrying a LHON mutation. Fibroblasts from LHON-affected patients showed pharmacologically reversible endoplasmic reticulum stress. DOA project: Variations in the concentration of some lipids, glutamate and polar neuroprotective metabolites suggested pre-symptomatic alterations of the myelin sheath along with axonal metabolic dysfunction of the optic nerve in Opa1 +/-mice. A sexual dimorphism was also observed in the metabolome of the optic nerve. RLIP project: Preconditioning light seemed to elicit acute proteolysis and decreased NO production in the retina. Light stress was also related with lipid remodeling in the retina. A sexual dimorphism was also observed in the retina of control rats. Taken as a whole, our results show that the metabolomics approach is adapted and relevant for the study of the physiopathology of ocular diseases

Maternal smoking during pregnancy (MSDP) and postnatal secondhand smoking (SHS) are ongoing public health concerns that are associated with adverse child health outcomes, but not much is known about the molecular mechanisms. We investigated the association between MSDP and placental DNA methylation, and its link with reproductive outcomes through a meta-analysis. We identified 1224 differentially methylated CpGs in placenta, which were enriched for pathways related to inflammation, growth factors and vascularization. Moreover, the methylation of many of these CpGs was associated with gestational age and birth size. We also investigated the association between MSDP and postnatal SHS, and different molecular layers in children: blood DNA methylation and transcription, plasma proteins, and serum and urinary metabolites. In utero exposure, was only associated with child DNA methylation, confirming a persistent MSDP-related signature on the blood epigenome. However, this imprint was not mirrored in the transcriptome. In contrast, postnatal SHS was related to protein and metabolite levels, which are more dynamic, and likely reflecting short-term exposures. Of note, the MSDP-associated methylome signature was tissue-specific.
Fumar durant l’embaràs i l’exposició a tabac passiu de manera postnatal són preocupacions en la salut pública que estan associades a conseqüències adverses en la salut infantil, però poc se sap sobre els mecanismes moleculars. Hem investigat l’associació entre fumar durant l’embaràs i la metilació de l’ADN placentari, i la seva relació amb les conseqüències reproductives a través d’un meta-anàlisi. Hem identificat 1224 CpGs diferencialment metilats a placenta, enriquits en vies relacionades amb inflamació, factors de creixement i vascularització. A més a més, la metilació de molts CpGs està associada a l’edat gestacional i les mides al naixement. Hem investigat també l’associació entre fumar durant l’embaràs i l’exposició a tabac passiu de manera postnatal, i diferents capes moleculars en nens: metilació de l’ADN i transcripció en sang, proteïnes en plasma, metabòlits en sèrum i orina. L’exposició uterina, estava només associada amb la metilació de l’ADN del nen, confirmant un efecte persistent del patró de l’epigenoma en sang. Tot i això, aquesta empremta no es veu reflectida en el transcriptoma. En canvi, l’exposició a tabac passiu estava relacionada amb proteïnes i metabòlits, que són més dinàmics, i reflecteixen efectes a més curt termini. Hem observat també que el patró de metilació és específic de teixit.

Les cancers du sein et de la prostate sont parmi les cancers ayant la plus forte incidence dans le monde,notamment dans les pays occidentaux. Les principaux défis actuels sont d’améliorer la compréhension des relations nutrition/santé et l’identification des personnes à plus haut risque bien avant l’apparition du cancer afin de mettre en place des actions de préventions. De nombreux facteurs influencent la mise en place et la progression du cancer. Parmi eux, la nutrition apparaît comme un facteur clé, puisqu’il s’agit d’un facteur modifiable sur lequel il est possible d'agir via des interventions, il est donc essentiel d’évaluer sa contribution. Pour cela, une mesure précise de l'apport nutritionnel est nécessaire. La métabolomique permettant l’identification de potentiels biomarqueurs endogènes, exogènes et microbiens ouvre donc de nouvelles perspectives en épidémiologie nutritionnelle. A ce jour, encore très peu d’études ont investigué l’impact de l’alimentation globale sur le métabolisme et le risque de cancer du sein et de la prostate par profilage métabolomique. Dans le cadre de cette thèse, nous avons donc conduit des études cas-témoins nichées et transversales dans la cohorte SU.VI.MAX afin de mettre en évidence des signatures plasmatiques du risque de cancers du sein et de la prostate et de l’alimentation globale. Les échantillons plasmatiques ont été collectés à l’inclusion dans la cohorte et analysés par deux méthodes complémentaires : la spectrométrie de masse couplée à la chromatographie liquide et la résonance magnétique nucléaire du proton. Les habitudes alimentaires des participants ont été estimées grâce à des enregistrements alimentaires de 24h répétés et les données socio-démographiques et de mode de vie ont été obtenues grâce à des questionnaires autodéclarés. Ces recherches ont permis de mettre en évidence des métabolites endogènes et issus du métabolisme microbien associés à l’alimentation globale et également des biomarqueurs candidats d’une exposition alimentaire spécifique. Nous avons également identifié des métabolites associés au risque de cancers du sein et de la prostate, endogènes, exogènes et microbiens suggérant une perturbation métabolique jusqu’à 13 ans avant le diagnostic du cancer. Par ailleurs, l’alimentation semble jouer un rôle dans la variation des taux plasmatiques de certains métabolites permettant de discriminer les individus à plus haut risque de développer un cancer du sein ou de la prostate. Ces résultats devront être répliqués dans d’autres études indépendantes d’observation et d’intervention.A terme, l’identification de signatures métaboliques robustes du risque de cancers du sein et de la prostate, de l’impact de l’alimentation sur le métabolisme et la cancérogenèse et de l’apport alimentaire pourraient permettre de contribuer à l’amélioration de la compréhension des relations entre environnement et santé, de l’évaluation de l’exposition nutritionnelle voire à la mise en place de nouvelles recommandations en matière de santé publique en vue de la diminution de l’incidence de ces pathologies
Breast and prostate cancers are among the cancers with the highest incidence worldwide and notably in Western countries. The main current challenges lie in the improvement of understanding of nutrition/health relationships and in the identification of individuals at higher risk long before the development of overt cancer to set up prevention actions. A variety of factors exert an impact on the onset and progression of cancer. Among these, nutrition appears as a key factor, in that it can be modified and acted upon through interventions. It is therefore crucial to assess its contribution. For this purpose,detailed and accurate assessment of nutritional intake is essential. Metabolomics, allowing the identification of endogenous, exogenous and microbial biomarkers, opens new perspectives in nutritional epidemiology. So far, few have studies investigated the impact of overall diet on metabolism and risk of breast and prostate cancer through metabolomic profiling. As part of this thesis, we conducted nested case-controls and cross-sectional studies within the SU.VI.MAX cohort to highlight plasma signatures of breast and prostate cancer risks and of overall diet. Plasma samples were collected at baseline and were analysed using two complementary methods : mass spectrometry coupled with liquid chromatography and proton nuclear magnetic resonance. Participants dietary habits were estimated using repeated 24h dietary records and socio-demographic and lifestyle data were collected from self-administered questionnaires.These investigations highlighted endogenous and microbial metabolites associated with overall diet as well as candidate biomarkers of specific dietary exposures. We also identified endogenous, exogenous and microbial metabolites associated with breast and prostate cancers risk suggesting a metabolic disruption up to 13 years before cancer diagnostic. Furthermore, diet appears to be implicated in the variation in plasma levels of some metabolites discriminating individuals at higher risk of developing breast or prostate cancers. These results need to be replicated in future independent observational and interventional studies. In the future, the identification of robust metabolic signatures of breast and prostate cancers risk, of the impact of diet on metabolism and carcinogenesis, and food intake would contribute to better understand health and environment relationships, to better estimate nutritional exposure or even to contribute to the set-up of new public health recommendations in order to reduce the incidence of these pathologies

La disponibilité de sources de protéines pour l’alimentation humaine est une préoccupation majeure du fait des transitions démographiques, économiques et nutritionnelles mondiales. Les apports en protéines doivent couvrir les besoins en neuf acides aminés considérés comme indispensables (AAI). Il est important que leur besoin soit couvert pour éviter des situations de déséquilibre du métabolisme protéique et énergétique. Différentes études ont été développées afin de déterminer ce besoin : bilan azoté, méthode factorielle, et méthodes utilisant les isotopes stables. Cependant, ces méthodes sont lourdes et invasives et les valeurs de besoins obtenues présentes des différences importantes. Il est donc nécessaire de développer des approches plus précises et non invasives, telle que la métabolomique, recommandée par la FAO. Les objectifs de cette thèse sont d’évaluer l’impact de la déficience en protéines et en AAI (lysine et thréonine) sur le métabolisme protéique et énergétique et d’identifier des marqueurs de la déficience de ces deux acides aminés chez le rat en croissance. Des niveaux sévères de déficience (85% ; 75%) en protéines et en lysine et thréonine diminuent le poids et la masse maigre et augmentent la prise alimentaire. Ces effets sont associés à une diminution de la synthèse protéique. et à une augmentation du métabolisme énergétique pour les faibles apports en protéines. Ces effets semblent médiés par FGF21. Les analyses de métabolomiques dans les urines révèlent que des variations du pipecolate et de la taurine signent respectivement une déficience en lysine et en thréonine
The availability of protein sources for human nutri-tion is a major concern due to global demographics, economics and nutritional transitions. Protein intakes must cover the need for nine indispensable amino acids (IAA). It is important that this need is met in order to avoid situations of protein and energy me-tabolism imbalance. Various studies have been de-veloped to determine this need: nitrogen balance, the factorial method, and methods using stable iso-topes. However, these methods are difficult and invasive, and the obtained values of needs present significant differences. It is therefore necessary to develop more precise and non-invasive approaches, such as metabolomics, as recommended by the FAO.The objectives of this thesis are to assess the impact of protein and IAA (lysine and threonine) deficiency on protein and energy metabolism and to identify markers of deficiency for these two amino acids in the growing rat. Severe levels of deficiency (85%; 75%) in protein and lysine and threonine decrease weight and lean mass and increase food intake. These effects are associated with a decrease in protein synthesis and an increase in energy metabolism in low protein diets. These effects seems to be mediated by FGF21. Analyses of metabolomics in urine show that variations in pipecolate and taurine indicate lysine and threonine deficiencies, respectively

Les huiles essentielles (HE) de lavande et de lavandin appartiennent au patrimoine de la région méditerranéenne. Comme tout produit naturel ou synthétique à valeur ajoutée, ces HE doivent être contrôlées pour justifier la qualité du produit. Ce contrôle qualité nécessite la mise en place d'une méthodologie analytique fiable. Dans cette étude, une nouvelle approche utilisant les techniques spectroscopiques et chromatographiques et le traitement de données à l'aide d'outils chimiométriques permet de différencier les HE de lavande et de lavandin par origine variétale. Cette discrimination s'effectue grâce à leur empreinte spectrale ou chromatographique. L’intérêt d’utiliser la chromatographie chirale associée à la détection polarimétrique et de la signature chiroptique acquise a également été étudié. Cette étude nous a permis d’identifier des marqueurs métabolomiques qui sont primordiaux pour caractériser les variétés. Les résultats montrent qu'il est possible de discriminer les HE de lavande et de lavandin selon leurs variétés avec une bonne justesse sur l'ensemble des techniques utilisées
Lavender and lavandin essential oils (EOs) belong to the heritage of the Mediterranean region. Like any natural or synthetic product with an added value, these EOs must be controlled to justify the quality of the product. This quality control requires the establishment of a reliable analytical methodology. In this study, a new approach using spectroscopic and chromatographic techniques for data processing associated to chemometric tools allows to discriminate lavender and lavandin EOs. This discrimination is carried out thanks to their spectroscopic or chromatographic fingerprints. The interest to use the chiral chromatography combined with polarimetric detection and of acquired chiroptical signature was also studied. This methodology has allowed us to identify metabolomic markers which are paramount to characterize the varieties. The results show that it is possible to discriminate the lavender and lavandin EOs according to their varieties with good accuracy on all of the techniques used

Les progrès de la recherche sur le cancer du sein dépendent de la disponibilité d’outils appropriés, comme les lignées cellulaires qui peuvent être implantées chez des souris immunocompétentes. La souche de souris C57Bl/6 est la plus étudiée et c’est la seule pour laquelle certaines variantes génétiques sont disponibles. Étant donné qu'aucune lignée cellulaire de carcinome mammaire à récepteurs hormonaux positifs de souche C57Bl/6 n'est disponible, nous avons décidé d'établir des lignées cellulaires de ce type. Nous avons induit des cancers du sein chez des souris C57BL/6 femelles en utilisant un analogue synthétique de la progestérone combiné à un agent endommageant l'ADN. Des lignées cellulaires ont été établies à partir de ces tumeurs et sélectionnées pour leur positivité au niveau du double récepteur (estrogène + progestérone), ainsi que pour leur transplantabilité chez les femelles C57BL/6. Parmi plusieurs lignées, une lignée cellulaire, que nous avons appelée MD5, remplissait ces critères et a permis l'établissement de tumeurs mal différenciées et très prolifératives. Ces tumeurs ont réduit leur croissance (sans toutefois régresser) lors du traitement par des antagonistes des récepteurs d’œstrogènes, ainsi que par une chimiothérapie à base d'anthracylines. Cependant, ce dernier effet n'a pas été influencé par la déplétion des lymphocytes T et, en outre, ces tumeurs n'ont pas répondu au blocage de PD-1, ce qui suggère que les tumeurs MD5 sont immunologiquement froides. En conclusion, les cellules MD5, dérivées des animaux C57BL/6, constituent un modèle de cancer du sein à récepteurs hormonaux positifs de mauvais pronostic
Progress in breast cancer research relies on the availability of suitable cell lines that can be implanted in immunocompetent laboratory mice. The best explored mouse strain, C57Bl/6, is also the only one for which multiple genetic variants are available. Driven by the fact that no hormone receptor-positive C57Bl/6-derived mammary carcinoma cell lines are available, we decided to establish such cell lines. Breast cancers were induced in female C57BL/6 mice using a synthetic progesterone analogue combined with a DNA damaging agent. Cell lines were established from these tumors and selected for dual (estrogen + progesterone) receptor positivity, as well as transplantability into C57BL/6 females. One cell line, which we called MD5,fulfilled these criteria and allowed for the establishment of poorly differentiated, highly proliferative, immune cold tumors. Such tumors reduced their growth (though did not regress) upon treatment with estrogen receptor antagonists, as well as with anthracyline-based chemotherapy. However, the latter effect was not influenced by T cell depletion and MD tumors failed to respond to PD-1 blockade, suggesting that they are immunologically cold. In conclusion, C57BL/6-derived MD5 cells constitute a model of poor prognosis hormone receptor-positive breast cancer

Les huiles essentielles (HE) de lavande et de lavandin appartiennent au patrimoine de la région méditerranéenne. Comme tout produit naturel ou synthétique à valeur ajoutée, ces HE doivent être contrôlées pour justifier la qualité du produit. Ce contrôle qualité nécessite la mise en place d'une méthodologie analytique fiable. Dans cette étude, une nouvelle approche utilisant les techniques spectroscopiques et chromatographiques et le traitement de données à l'aide d'outils chimiométriques permet de différencier les HE de lavande et de lavandin par origine variétale. Cette discrimination s'effectue grâce à leur empreinte spectrale ou chromatographique. L’intérêt d’utiliser la chromatographie chirale associée à la détection polarimétrique et de la signature chiroptique acquise a également été étudié. Cette étude nous a permis d’identifier des marqueurs métabolomiques qui sont primordiaux pour caractériser les variétés. Les résultats montrent qu'il est possible de discriminer les HE de lavande et de lavandin selon leurs variétés avec une bonne justesse sur l'ensemble des techniques utilisées
Lavender and lavandin essential oils (EOs) belong to the heritage of the Mediterranean region. Like any natural or synthetic product with an added value, these EOs must be controlled to justify the quality of the product. This quality control requires the establishment of a reliable analytical methodology. In this study, a new approach using spectroscopic and chromatographic techniques for data processing associated to chemometric tools allows to discriminate lavender and lavandin EOs. This discrimination is carried out thanks to their spectroscopic or chromatographic fingerprints. The interest to use the chiral chromatography combined with polarimetric detection and of acquired chiroptical signature was also studied. This methodology has allowed us to identify metabolomic markers which are paramount to characterize the varieties. The results show that it is possible to discriminate the lavender and lavandin EOs according to their varieties with good accuracy on all of the techniques used

La reperfusion précoce à la phase aiguë de l’infarctus du myocarde améliore le pronostic des patients. Elle induit cependant une partie des lésions d’ischémie-reperfusion (I/R) liées à l’activation de mécanismes métaboliques et inflammatoires. Le conditionnement ischémique à distance (RIC) d’une part et les approches pharmacologiques d’autre part, constituent un espoir dans la prévention des lésions de reperfusion contre lesquelles nous ne disposons pas de traitement validé chez l’homme. L’objectif de cette thèse était d’explorer ces stratégies de cardioprotection à la phase aiguë de l’infarctus du myocarde. Grace à une approche métabolomique, nous avons identifié la kynurénine et glycine comme métabolites associés au RIC sur des plasmas de rats et confirmés dans une cohorte de patients. Nous avons ainsi validé in vivo l’action bénéfique de la kynurénine et de la glycine dans un modèle d’infarctus du myocarde. Nous avons ensuite étudié la modulation de la voie des kynurénines dans la cardioprotection induite par le RIC. Nous avons observé une activation de la voie de synthèse du NAD+ associée à une déacétylation des protéines mitochondriales hépatiques. Dans un dernier travail réalisé in vivo et in vitro, nous avons étudié le rôle cardioprotecteur de la colchicine dans l’I/R et analysé ces effets sur la modulation de l’inflammation et l’activation des voies de survie
The introduction of early reperfusion in the acute phase of myocardial infarction has improved the prognosis of patients. However, it induces irreversible damages called ischemia-reperfusion (I / R) injury followed by myocardial metabolic and inflammatory disorders. Remote ischemic conditioning (RIC) on the one hand and pharmacological approaches on the other hand, constitute a hope in the prevention of reperfusion injury against which we do not have validated treatment in humans.The aim of this thesis was to explore cardioprotection strategies in the acute phase of myocardial infarction. Using a metabolomics approach, we identified kynurenine and glycine as RIC-associated metabolites in rat plasmas and confirmed in a cohort of patients. We have also validated in vivo the beneficial effect of kynurenine and glycine in a model of myocardial infarction. We then studied the modulation of the kynurenine pathway in RIC-induced cardioprotection. We observed an activation of the NAD + synthesis pathway associated with deacetylation of hepatic mitochondrial proteins. In a last work carried out in vivo and in vitro, we studied the cardioprotective role of colchicine in I / R and analyzed its immunomodulatory effect and activation of survival pathways

Metabolic Syndrome (MetS) is an obesity-related disorder that predisposes an individual to several life-threatening diseases such as cardiovascular disease, hypertension and type 2 diabetes mellitus. The diagnosis of metabolic syndrome is based on the presence of at least 3 of the following 5 risk factors: elevated triglycerides, high blood pressure, high blood glucose, low HDL cholesterol and central adiposity. However, the biochemical mechanisms underlying the contribution of these irregularities are not fully understood. Currently, there is a need to better characterize MetS. Irregularity of lipid abundances, dyslipidemia, is known to be associated with MetS. However, little is known about the link between plasma phospholipids and human metabolic syndrome. In this study, mass spectrometry-based metabolomic approaches were employed using ultrahigh-resolution FTICR mass spectrometry to qualitatively analyze human plasma phospholipids and high-resolution QTOF mass spectrometry to quantitatively detect differences in the human plasma phospholipid profiles from 10 clinically-diagnosed metabolic syndrome patients and 8 lean healthy controls. The results point to the existence of a phospholipid signature of MetS. Five of the top twenty phospholipids contributing most to the difference in phospholipid abundance between the MetS and control group were identified using accurate mass-based database searching and MS/MS for structural confirmation. Relative differences in phospholipid abundances between MetS and controls for all top 20 phospholipids were shown to be statistically significant. These results may aid biomarker discovery and the accurate evaluation and prevention of diseases associated with dyslipidemia including human metabolic syndrome.
Graduate

碩士
輔仁大學
營養科學系碩士班
107
Lung cancer is the leading cause of cancer death, 85% is non-small cell lung cancer (NSCLC). One carbon nutritional status was associated with lung cancer risk, folate malnutrition may reprogram Warburg metabolism, induced cancer stemness releasing lactate and changed microenvironment to motivate malignant tumor progression. However, tumor energy metabolite – lactate in Taiwan NSCLC patients may be related to one carbon nutrition which is unknown. Thus, it is necessary to evaluate NSCLC patients’ one carbon nutritional status, correlation between lactate and tumor metabolites. We recruited 54 lung cancer patients in NTUH Division of Thoracic Surgery, dietary intake was analyzed by semi-quantitative one carbon nutrient food frequency questionnaire, blood was analyzed for lactate and one carbon nutrients; 12 pairs tumor/non-tumor tissues were analyzed by metabolomics. We found that tumor metabolomic markers, such as lactate, ADP, AMP, and GDP were significantly higher than non-tumor tissues, but glucose was upside down, which meant that tumor extremely required energy. Other study found that lung tumor would use lactate as fuel; Through the correlation heat map, which assumed that lactate tended to TCA cycle. Compared several metabolites/glucose ratios at different stages, we were found that lactate/glucose ratio was significantly higher in late stage than early stage of tumor. Compared blood values of different stages, we found that lactate levels in tumor late stage was significantly higher than benign tumor (p<.05), it was also found that while tumor stages increased, folate intake, vitamin B12 intake, RBC folate levels, plasma vitamin B12 levels significantly increased too (p<.05). Compared the relationship between lactate and methyl nutrients, we found that RBC folate, plasma vitamin B12 had significantly positive correlation with plasma lactate (r=0.329, p=0.015; r=0.453, p=0.004). According to the above, we assumed that lactate was associated with methyl nutrition. However, tumor’s nutrition came from the microenvironment provided nutrients from blood vessel, therefore we needed to understand methyl nutritional status of NSCLC patients in Taiwan. Blood folate malnutrition rate 18-20%, folate intake malnutrition rate was 1/3, and 7.5% was exceeded UL. Blood vitamin B12 malnutrition rate was 11.1%, which was exceeded the upper limit level, and vitamin B12 malnutrition rate was 1/5. Therefore, more researches were needed to understand the effect between methyl nutrients and lung cancer, which could give the correct direction of methyl nutritional supplementation in clinical.