Magnetic resonance imaging (MRI) reporter genes produce MRI signal in response to the molecular environment of the cells in which they are expressed. With an MRI scanner, the signal is detected and used to produce an image of the cells. We hypothesized that the magnetotactic bacterial gene mms6 has the potential to function as an MRI reporter gene. Magnetotactic bacteria produce magnetic iron oxide crystals in intracellular organelles called magnetosomes. mms6 encodes an iron-binding, magnetosome membrane protein Mms6, which plays a role in regulating the size and shape of the iron oxide crystals found within the magnetosomes. To test our hypothesis, several mammalian cell lines were transfected with mms6, and mms6-positive clones were genetically engineered. We then used MRI to image these clones in vitro. When the cells were incubated with iron-supplemented culture media, the mms6-positive clones produced more MRI image contrast than mms6-negative cells. Through a systematic process of elimination, the mms6-positive clone that generated the most in vitro MRI image contrast was identified. This clone, named 9L4S, was composed of mms6-positive rat glioma (9L) cells and was used for intracellular iron studies and in vivo imaging. The results of electron microscopy and optical emission spectrometry support the theory that mms6-positive clones enhance MRI image contrast due to an increase in intracellular iron. The main objective of this research was to assess the ability of mms6 to function as an in vivo MRI reporter gene, so a flank tumor animal model was created. Without any exogenous iron supplementation, tumors composed of mms6-positive cells produced greater negative contrast on an MRI image than mms6-negative cells. These results demonstrate that mms6 can be considered for use in studies requiring an MRI reporter gene.

Les récents développements en RMN et IRM ont eu un impact spectaculaire sur l’imagerie médicale et aident à appréhender de manière fonctionnelle et non invasive de nombreux mécanismes cérébraux. L’imagerie par RMN de perfusion a notamment une importance primordiale en neuroimagerie clinique dans la caractérisation de nombreux désordres cérébrovasculaires (tumeurs cérébrales, AVC). Cependant il n’existe pour le moment aucune technique qui quantifie de manière non-invasive le bon fonctionnement du réseau microvasculaire cérébral. Cette thèse se concentre sur l’utilisation de la séquence FENSI en imagerie préclinique à haut et ultra haut champ magnétique pour caractériser et quantifier la dynamique des flux microvasculaires dans le cerveau du rat. Nous présentons les enjeux de l’IRM de perfusion en neuroimagerie, ainsi que les contraintes associées aux méthodes conventionnelles – méthode de marquage de spin artériel (ASL) et IRM dynamique de contraste de susceptibilité magnétique (DSC-MRI) – dont la technique FENSI peut d’affranchir. Cependant d’autres problèmes sont adressés. Les images acquises avec FENSI et pondérées en flux peuvent être contaminées par des effets de transferts d’aimantation (MT) qui empêchent de quantifier le débit sanguin cérébral. Une première technique de correction de ces effets par post-traitement est proposée. Nous dérivons les premières cartes de flux sanguin cérébral chez le rat à 7 tesla. Une seconde approche est considérée, où les effets MT dans les images acquises avec et sans saturation des spins circulant dans le réseau capillaire (tag/control) se compensent. Cette seconde approche permet une vraie quantification non-invasive du flux sanguin cérébral (CBFlux) in-vivo. Le réseau microvasculaire est caractérisé via FENSI à différents stages du développement tumoral dans un modèle de gliosarcome cérébral (9L) chez le rat. Les mesures de flux mettent en évidence une forte hétérogénéité de développement vasculaire des gliosarcomes peu avancés (diamètre inférieur à 3 mm). A un stade avancé, le flux sanguin est significativement plus faible dans la tumeur (-40 %) que dans le sous cortex, en bon accord avec la littérature sur ce type de gliosarcome. De plus, les cartes paramétriques de flux permettent de distinguer différents compartiments dans la tumeur. Une comparaison avec une méthode de marquage endothélial sur coupes histologiques suggère que le flux sanguin calculé avec FENSI est corrélé avec la concentration locale en micro-vaisseaux. Nous avons également tentés d’évaluer la dynamique temporelle de la réponse vasculaire à un stimulus et d’appliquer FENSI à l’IRM fonctionnelle (IRMf). Nous avons mis en place un protocole d’IRMf chez le rat à 7 tesla et caractérisé la réponse hémodynamique obtenue par contraste BOLD. A 7 tesla la technique FENSI souffre d’un faible SNR temporel et semble plus adaptée pour quantifier des changements métaboliques associées à de longues plages temporelles. L’implémentation de la séquence à ultra-haut champ (17.2 tesla) donne lieu à de sérieux espoirs en IRMf. De plus, nous mettons en évidence à 17.2 tesla des contrastes spécifiques à l’utilisation de différents anesthésiques utilisés en routine à l’hôpital. La méthode que nous avons mise en place peut être sensibilisée à de nombreuses vitesses, augmentant le nombre de ses applications potentielles. Ainsi, un choix judicieux de paramètres permet d’explorer le volume sanguin ou l’orientation du débit ciblé. Les forces et faiblesses de la méthode sont détaillées. L’utilisation de FENSI n’est en général pas justifiée par un gain de signal par rapport à des séquences ASL optimisées, mais bénéficie de l’actuelle montée en champ des imageurs cliniques pour être étudiée. Les applications potentielles varient de l’IRMf et l’imagerie de diffusion au suivi pharmacologique et diagnostic de désordres cérébrovasculaires, dont l’étude via ASL est limitée dû à l’allongement des temps de transit sanguins. / The discoveries, implementations and developments of NMR and MRI have had a major impact in medical imaging. Compared to other imaging modalities (PET, SPECT, CT), current MRI research helps to further and better understand the inner mechanisms of the human body in a less invasive manner. In clinical neuroimaging, perfusion MRI is of spectacular importance to study cerebrovascular diseases and cancer. However, at the moment, there is no perfusion MRI sequence that allows for a complete, non-invasive and precise quantification of microvascular flow dynamics. This work focuses on the use of the recently introduced Flow Enhanced Signal Intensity method (FENSI) to characterize and quantify vasculature at capillary level, at high and ultra high magnetic field (7 and 17.2 tesla). For that purpose, the possible quantification of blood flux with FENSI is explored in vivo. The combination of flux quantification and flow-enhanced signal (compared to Arterial Spin Labeling) can make of FENSI an ideal method to characterize in a complete non-invasive way the brain microvasculature. After removal of magnetization transfer (MT) effects, the blood flow dynamics are studied with FENSI in a very aggressive and propagative rat brain tumor model: the 9L gliosarcoma. The objective is to assess whether FENSI is suitable for a longitudinal non-invasive characterization of microvascular changes associated with tumor growth. The results obtained with FENSI are compared with literature on 9L perfusion and immuno-histochemistry. In the first paper published on FENSI, a first glance was also casted on the potential of the flow enhanced technique when applied to fMRI. The results obtained at the time were contaminated by MT effects. With the implementation of a new MT-free FENSI technique, the possibility to map the brain cerebral functioning based on a quantitative physiological parameter (CBFlux) more directly related to neuronal activity than the usual BOLD signal is within reach. At ultra high field, the influence of different anesthetics on the rat brain microvascular network and BOLD contrast is also considered. After many developments around the FENSI technique, the method is compared to classical ASL and DSC perfusion MRI sequences. The strengths and weaknesses of the FENSI method, its characteristics, ‘precautions for use’, and potential main applications are detailed and discussed.

The discoveries, implementations and developments of NMR and MRI have had a major impact in medical imaging. Compared to other imaging modalities (PET, SPECT, CT), current MRI research helps to further and better understand the inner mechanisms of the human body in a less invasive manner. In clinical neuroimaging, perfusion MRI is of spectacular importance to study cerebrovascular diseases and cancer. However, at the moment, there is no perfusion MRI sequence that allows for a complete, non-invasive and precise quantification of microvascular flow dynamics. This work focuses on the use of the recently introduced Flow Enhanced Signal Intensity method (FENSI) to characterize and quantify vasculature at capillary level, at high and ultra high magnetic field (7 and 17.2 tesla). For that purpose, the possible quantification of blood flux with FENSI is explored in vivo. The combination of flux quantification and flow-enhanced signal (compared to Arterial Spin Labeling) can make of FENSI an ideal method to characterize in a complete non-invasive way the brain microvasculature. After removal of magnetization transfer (MT) effects, the blood flow dynamics are studied with FENSI in a very aggressive and propagative rat brain tumor model: the 9L gliosarcoma. The objective is to assess whether FENSI is suitable for a longitudinal non-invasive characterization of microvascular changes associated with tumor growth. The results obtained with FENSI are compared with literature on 9L perfusion and immuno-histochemistry. In the first paper published on FENSI, a first glance was also casted on the potential of the flow enhanced technique when applied to fMRI. The results obtained at the time were contaminated by MT effects. With the implementation of a new MT-free FENSI technique, the possibility to map the brain cerebral functioning based on a quantitative physiological parameter (CBFlux) more directly related to neuronal activity than the usual BOLD signal is within reach. At ultra high field, the influence of different anesthetics on the rat brain microvascular network and BOLD contrast is also considered. After many developments around the FENSI technique, the method is compared to classical ASL and DSC perfusion MRI sequences. The strengths and weaknesses of the FENSI method, its characteristics, 'precautions for use', and potential main applications are detailed and discussed.