Vincent Lebon

Quantitative susceptibility map reconstruction from MR phase data using bayesian regularization: validation and application to brain imaging.

The diagnosis of many neurologic diseases benefits from the ability to quantitatively assess iron in the brain. Paramagnetic iron modifies the magnetic susceptibility causing magnetic field inhomogeneity in MRI. The local field can be mapped using the MR signal phase, which is discarded in a typical image reconstruction. The calculation of the susceptibility from the measured magnetic field is an ill‐posed inverse problem. In this work, a bayesian regularization approach that adds spatial priors from the MR magnitude image is formulated for susceptibility imaging. Priors include background regions of known zero susceptibility and edge information from the magnitude image. Simulation and phantom validation experiments demonstrated accurate susceptibility maps free of artifacts. The ability to characterize iron content in brain hemorrhage was demonstrated on patients with cavernous hemangioma. Additionally, multiple structures within the brain can be clearly visualized and characterized. The technique introduces a new quantitative contrast in MRI that is directly linked to iron in the brain. Magn Reson Med, 2010.

Reactive Astrocytes Overexpress TSPO and Are Detected by TSPO Positron Emission Tomography Imaging

Astrocytes and microglia become reactive under most brain pathological conditions, making this neuroinflammation process a surrogate marker of neuronal dysfunction. Neuroinflammation is associated with increased levels of translocator protein 18 kDa (TSPO) and binding sites for TSPO ligands. Positron emission tomography (PET) imaging of TSPO is thus commonly used to monitor neuroinflammation in preclinical and clinical studies. It is widely considered that TSPO PET signal reveals reactive microglia, although a few studies suggested a potential contribution of reactive astrocytes. Because astrocytes and microglia play very different roles, it is crucial to determine whether reactive astrocytes can also overexpress TSPO and yield to a detectable TSPO PET signal in vivo. We used a model of selective astrocyte activation through lentiviral gene transfer of the cytokine ciliary neurotrophic factor (CNTF) into the rat striatum, in the absence of neurodegeneration. CNTF induced an extensive activation of astrocytes, which overexpressed GFAP and become hypertrophic, whereas microglia displayed minimal increase in reactive markers. Two TSPO radioligands, [18F]DPA-714 [N,N-diethyl-2-(2-(4-(2-[18F]fluoroethoxy)phenyl)-5,7-dimethylpyrazolo[1,5-a]pyrimidin-3-yl)acetamide] and [11C]SSR180575 (7-chloro-N,N-dimethyl-5-[11C]methyl-4-oxo-3-phenyl-3,5-dihydro-4H-pyridazino[4,5-b]indole-1-acetamide), showed a significant binding in the lenti-CNTF-injected striatum that was saturated and displaced by PK11195 [N-methyl-N-(1-methylpropyl)-1-(2-chlorophenyl)-isoquinoline-3-carboxamide]. The volume of radioligand binding matched the GFAP immunopositive volume. TSPO mRNA levels were significantly increased, and TSPO protein was overexpressed by CNTF-activated astrocytes. We show that reactive astrocytes overexpress TSPO, yielding to a significant and selective binding of TSPO radioligands. Therefore, caution must be used when interpreting TSPO PET imaging in animals or patients because reactive astrocytes can contribute to the signal in addition to reactive microglia.

SIMULTANEOUS MEASUREMENT OF PERFUSION AND OXYGENATION CHANGES USING A MULTIPLE GRADIENT-ECHO SEQUENCE: APPLICATION TO HUMAN MUSCLE STUDY

We have developed a magnetic resonance imaging (MRI) technique based on a multiple gradient-echo sequence designed to probe perfusion and oxygenation simultaneously within skeletal muscle. Processing of the images acquired at successive echo times (TEs) generates two functional maps: one of the signal intensity (SI) extrapolated to zero echo time, which is sensitive to perfusion; and a second one of R2*, which reflects oxygenation. An advantage of the processing procedure lies in the selection of tissue of interest through the profile of T2* decay, leading to automatic rejection of pixels containing small vessels. This allows a more specific assessment of tissue perfusion and oxygenation. This technique was demonstrated successfully during post-ischemic reactive hyperemia in human calf. A perfusion peak of 123 mL x 100 g(-)1 x min(-1) was measured immediately after ischemia, whereas R2* value showed an 11.5% decrease at the same time, essentially reflecting blood oxygenation changes. Differences in the time courses of reperfusion and re-oxygenation were observed, oxygenation presenting a slower recovery. The mechanisms responsible for such a differential dynamic response are discussed.

Decreased TCA cycle rate in the rat brain after acute 3-NP treatment measured by in vivo 1H-[13C] NMR spectroscopy.

Inhibition of succinate dehydrogenase (SDH) by the mitochondrial toxin 3‐nitropropionic acid (3‐NP) has gained acceptance as an animal model of Huntingtons disease. In this study 13C NMR spectroscopy was used to measure the tricarboxylic acid (TCA) cycle rate in the rat brain after 3‐NP treatment. The time course of both glutamate C4 and C3 13C labelling was monitored in vivo during an infusion of [1‐13C]glucose. Data were fitted by a mathematical model to yield the TCA cycle rate (Vtca) and the exchange rate between α‐ketoglutarate and glutamate (Vx). 3‐NP treatment induced a 18% decrease in Vtca from 0.71 ± 0.02 µmol/g/min in the control group to 0.58 ± 0.02 µmol/g/min in the 3‐NP group (p < 0.001). Vx increased from 0.88 ± 0.08 µmol/g/min in the control group to 1.33 ± 0.24 µmol/g/min in the 3‐NP group (p < 0.07). Fitting the C4 glutamate time course alone under the assumption that Vx is much higher than Vtca yielded Vtca=0.43 µmol/g/min in both groups. These results suggest that both Vtca and Vx are altered during 3‐NP treatment, and that both glutamate C4 and C3 labelling time courses are necessary to obtain a reliable measurement of Vtca.

Influence of vascular filling and perfusion on BOLD contrast during reactive hyperemia in human skeletal muscle

Mechanisms generating BOLD contrast are complex and depend on parameters that are prone to large variations, in particular in skeletal muscle. Here, we simultaneously measured perfusion by ASL, and BOLD response in the calf muscle of 6 healthy volunteers during post‐ischemic reactive hyperemia. We tested whether the relation between the two was altered for varying degrees of leg vascular replenishment induced by prior positioning of the leg at different heights relative to the heart. We found that the BOLD response depended on perfusion, but also on the degree of repletion of leg blood vessels. We conclude that simultaneous determination of perfusion by ASL is important to identify the mechanisms underlying BOLD contrast in the skeletal muscle. Magn Reson Med, 2006.

Neuron–astrocyte interactions in the regulation of brain energy metabolism: a focus on NMR spectroscopy

An adequate and timely production of ATP by brain cells is of cardinal importance to support the major energetic cost of the rapid processing of information via synaptic and action potentials. Recently, evidence has been accumulated to support the view that the regulation of brain energy metabolism is under the control of an intimate dialogue between astrocytes and neurons. In vitro studies on cultured astrocytes and in vivo studies on rodents have provided evidence that glutamate and Na+ uptake in astrocytes is a key triggering signal regulating glucose use in the brain. With the advent of NMR spectroscopy, it has been possible to provide experimental evidence to show that energy consumption is mainly devoted to glutamatergic neurotransmission and that glutamate‐glutamine cycling is coupled in a ∼ 1 : 1 molar stoichiometry to glucose oxidation, at least in the cerebral cortex. This improved understanding of neuron–astrocyte metabolic interactions offers the potential for developing novel therapeutic strategies for many neurological disorders that include a metabolic deficit.

Multimodal neuroimaging provides a highly consistent picture of energy metabolism, validating 31P MRS for measuring brain ATP synthesis

Neuroimaging methods have considerably developed over the last decades and offer various noninvasive approaches for measuring cerebral metabolic fluxes connected to energy metabolism, including PET and magnetic resonance spectroscopy (MRS). Among these methods, 31P MRS has the particularity and advantage to directly measure cerebral ATP synthesis without injection of labeled precursor. However, this approach is methodologically challenging, and further validation studies are required to establish 31P MRS as a robust method to measure brain energy synthesis. In the present study, we performed a multimodal imaging study based on the combination of 3 neuroimaging techniques, which allowed us to obtain an integrated picture of brain energy metabolism and, at the same time, to validate the saturation transfer 31P MRS method as a quantitative measurement of brain ATP synthesis. A total of 29 imaging sessions were conducted to measure glucose consumption (CMRglc), TCA cycle flux (VTCA), and the rate of ATP synthesis (VATP) in primate monkeys by using 18F-FDG PET scan, indirect 13C MRS, and saturation transfer 31P MRS, respectively. These 3 complementary measurements were performed within the exact same area of the brain under identical physiological conditions, leading to: CMRglc = 0.27 ± 0.07 μmol·g−1·min−1, VTCA = 0.63 ± 0.12 μmol·g−1·min−1, and VATP = 7.8 ± 2.3 μmol·g−1·min−1. The consistency of these 3 fluxes with literature and, more interestingly, one with each other, demonstrates the robustness of saturation transfer 31P MRS for directly evaluating ATP synthesis in the living brain.

Isoflurane strongly affects the diffusion of intracellular metabolites, as shown by 1H nuclear magnetic resonance spectroscopy of the monkey brain

Isoflurane is a volatile anesthetic commonly used for animal studies. In particular, diffusion nuclear magnetic resonance (NMR) spectroscopy is frequently performed under isoflurane anesthesia. However, isoflurane is known to affect the phase transition of lipid bilayer, possibly resulting in increased permeability to metabolites. Resulting decreased restriction may affect metabolite apparent diffusion coefficient (ADC). In the present work, the effect of isoflurane dose on metabolite ADC is evaluated using diffusion tensor spectroscopy in the monkey brain. For the five detected intracellular metabolites, the ADC exhibits a significant increase when isoflurane dose varies from 1% to 2%: 13%±8% for myo-inositol, 14%±13% for total N-acetyl-aspartate, 20%±18% for glutamate, 27%±7% for total creatine and 53%±17% for total choline. Detailed analysis of ADC changes experienced by the five different metabolites argues in favor of facilitated metabolite exchange between subcellular structures at high isoflurane dose. This work strongly supports the idea of metabolite diffusion in vivo being significantly restricted in subcellular structures at long diffusion time, and provides new insights for interpreting ADC values as measured by diffusion NMR spectroscopy.

Anomalous diffusion of brain metabolites evidenced by diffusion-weighted magnetic resonance spectroscopy in vivo

Translational displacement of molecules within cells is a key process in cellular biology. Molecular motion potentially depends on many factors, including active transport, cytosol viscosity and molecular crowding, tortuosity resulting from cytoskeleton and organelles, and restriction barriers. However, the relative contribution of these factors to molecular motion in the cytoplasm remains poorly understood. In this work, we designed an original diffusion-weighted magnetic resonance spectroscopy strategy to probe molecular motion at subcellular scales in vivo. This led to the first observation of anomalous diffusion, that is, dependence of the apparent diffusion coefficient (ADC) on the diffusion time, for endogenous intracellular metabolites in the brain. The observed increase of the ADC at short diffusion time yields evidence that metabolite motion is characteristic of hindered random diffusion rather than active transport, for time scales up to the dozen milliseconds. Armed with this knowledge, data modeling based on geometrically constrained diffusion was performed. Results suggest that metabolite diffusion occurs in a low-viscosity cytosol hindered by ~2-μm structures, which is consistent with known intracellular organization.

Glycolysis versus TCA cycle in the primate brain as measured by combining 18F-FDG PET and 13C-NMR.

The glycolytic flux (cerebral metabolic rate of glucose CMRglc) and the TCA cycle flux (VTCA) were measured in the same monkeys by 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) and 13C NMR spectroscopy, respectively. Registration of nuclear magnetic resonance (NMR) and PET data were used for comparison of CMRglc and VTCA in the exact same area of the brain. Both fluxes were in good agreement with literature values (CMRglc 0.23 ± 0.03 μmol/g min, VTCA = 0.53 ± 0.13 μmol/gmin). The resulting [CMRglc/VTCA] ratio was 0.46 ± 0.12 (n = 5, mean ± s.d.), not significantly different from the 0.5 expected when glucose is the sole fuel that is completely oxidized. Our results provide a cross-validation of both techniques. Comparison of CMRglc with VTCA is in agreement with a metabolic coupling between the TCA cycle and glycolysis under normal physiologic conditions.