In Vivo Imaging Biomarkers in Mouse Models of Alzheimer's Disease: Are We Lost in Translation or Breaking Through?

Identification of biomarkers of Alzheimer's Disease (AD) is a critical priority to efficiently diagnose the patients, to stage the progression of neurodegeneration in living subjects, and to assess the effects of disease-modifier treatments. This paper addresses the development and usefulness of preclinical neuroimaging biomarkers of AD. It is today possible to image in vivo the brain of small rodents at high resolution and to detect the occurrence of macroscopic/microscopic lesions in these species, as well as of functional alterations reminiscent of AD pathology. We will outline three different types of imaging biomarkers that can be used in AD mouse models: biomarkers with clear translational potential, biomarkers that can serve as in vivo readouts (in particular in the context of drug discovery) exclusively for preclinical research, and finally biomarkers that constitute new tools for fundamental research on AD physiopathogeny

Evaluation of Human Interindividual Variation in Bioactivation of Estragole Using Physiologically Based Biokinetic Modeling

The present study investigates interindividual variation in liver levels of the proximate carcinogenic metabolite of estragole, 1′-hydroxyestragole, due to variation in two key metabolic reactions involved in the formation and detoxification of this metabolite, namely 1′-hydroxylation of estragole and oxidation of 1′-hydroxyestragole. Formation of 1′-hydroxyestragole is predominantly catalyzed by P450 1A2, 2A6, and 2E1, and results of the present study support that oxidation of 1′-hydroxyestragole is catalyzed by 17β-hydroxysteroid dehydrogenase type 2 (17β-HSD2). In a first approach, the study defines physiologically based biokinetic (PBBK) models for 14 individual human subjects, revealing a 1.8-fold interindividual variation in the area under the liver concentration-time curve (AUC) for 1′-hydroxyestragole within this group of human subjects. Variation in oxidation of 1′-hydroxyestragole by 17β-HSD2 was shown to result in larger effects than those caused by variation in P450 enzyme activity. In a second approach, a Monte Carlo simulation was performed to evaluate the extent of variation in liver levels of 1′-hydroxyestragole that could occur in the population as a whole. This analysis could be used to derive a chemical-specific adjustment factor (CSAF), which is defined as the 99th percentile divided by the 50th percentile of the predicted distribution of the AUC of 1′-hydroxyestragole in the liver. The CSAF was estimated to range between 1.6 and 4.0, depending on the level of variation that was taken into account for oxidation of 1′-hydroxyestragole. Comparison of the CSAF to the default uncertainty factor of 3.16 for human variability in biokinetics reveals that the default uncertainty factor adequately protects 99% of the populatio

Use of Physiologically Based Biokinetic (PBBK) Modeling to Study Estragole Bioactivation and Detoxification in Humans as Compared with Male Rats

The extent of bioactivation of the herbal constituent estragole to its ultimate carcinogenic metabolite 1′-sulfooxyestragole depends on the relative levels of bioactivation and detoxification pathways. The present study investigated the kinetics of the metabolic reactions of both estragole and its proximate carcinogenic metabolite 1′-hydroxyestragole in humans in incubations with relevant tissue fractions. Based on the kinetic data obtained a physiologically based biokinetic (PBBK) model for estragole in human was defined to predict the relative extent of bioactivation and detoxification at different dose levels of estragole. The outcomes of the model were subsequently compared with those previously predicted by a PBBK model for estragole in male rat to evaluate the occurrence of species differences in metabolic activation. The results obtained reveal that formation of 1′-oxoestragole, which represents a minor metabolic route for 1′-hydroxyestragole in rat, is the main detoxification pathway of 1′-hydroxyestragole in humans. Due to a high level of this 1′-hydroxyestragole oxidation pathway in human liver, the predicted species differences in formation of 1′-sulfooxyestragole remain relatively low, with the predicted formation of 1′-sulfooxyestragole being twofold higher in human compared with male rat, even though the formation of its precursor 1′-hydroxyestragole was predicted to be fourfold higher in human. Overall, it is concluded that in spite of significant differences in the relative extent of different metabolic pathways between human and male rat there is a minor influence of species differences on the ultimate overall bioactivation of estragole to 1′-sulfooxyestragol

Alzheimer disease models and human neuropathology: similarities and differences

Animal models aim to replicate the symptoms, the lesions or the cause(s) of Alzheimer disease. Numerous mouse transgenic lines have now succeeded in partially reproducing its lesions: the extracellular deposits of Aβ peptide and the intracellular accumulation of tau protein. Mutated human APP transgenes result in the deposition of Aβ peptide, similar but not identical to the Aβ peptide of human senile plaque. Amyloid angiopathy is common. Besides the deposition of Aβ, axon dystrophy and alteration of dendrites have been observed. All of the mutations cause an increase in Aβ 42 levels, except for the Arctic mutation, which alters the Aβ sequence itself. Overexpressing wild-type APP alone (as in the murine models of human trisomy 21) causes no Aβ deposition in most mouse lines. Doubly (APP × mutated PS1) transgenic mice develop the lesions earlier. Transgenic mice in which BACE1 has been knocked out or overexpressed have been produced, as well as lines with altered expression of neprilysin, the main degrading enzyme of Aβ. The APP transgenic mice have raised new questions concerning the mechanisms of neuronal loss, the accumulation of Aβ in the cell body of the neurons, inflammation and gliosis, and the dendritic alterations. They have allowed some insight to be gained into the kinetics of the changes. The connection between the symptoms, the lesions and the increase in Aβ oligomers has been found to be difficult to unravel. Neurofibrillary tangles are only found in mouse lines that overexpress mutated tau or human tau on a murine tau −/− background. A triply transgenic model (mutated APP, PS1 and tau) recapitulates the alterations seen in AD but its physiological relevance may be discussed. A number of modulators of Aβ or of tau accumulation have been tested. A transgenic model may be analyzed at three levels at least (symptoms, lesions, cause of the disease), and a reading key is proposed to summarize this analysis

Relationships between soil factors, Quercus robur health, Collybia fusipes root infection and Phytophthora presence

About sixty oaks (Quercus robur), with different status of crown health, located in three plots in north eastern France were assessed for their degree of infection by C. fusipes. Baiting of Phytophthora spp. from soil was carried out at different seasons in a sub sample of these oaks. The variability of soil characteristics, such as depth to waterlogging signs and depth to a gravel layer limiting rooting was evaluated. Soil factors were tightly correlated. Infection induced by C. fusipes was significantly more severe in non-waterlogged conditions. In two plots Phytophthora spp. were present at the base of the majority of trees investigated. Success of Phytophthora isolation was significantly linked with the season. The crown health status deteriorates with increasing severity of infection by C. fusipes and with the depth to waterlogging. Conversely, Phytophthora was not correlated with crown health status.Relations entre les facteurs du sol, l'état du houppier de Quercus robur, les infections racinaires induites par Collybia fusipes et la présence de Phytophthora. Le niveau d'infection induit par Collybia fusipes a été évalué sur environ 60 chênes pédonculés (Quercus robur) présentant différents états sanitaires, situés dans trois parcelles du nord-est de la France. Le piégeage des Phytophthora spp. présents dans le sol a été effectué sur un sous-échantillon de chênes à différentes saisons. La variabilité des facteurs du sol concernant la profondeur d'apparition des traces d'hydromorphie et la profondeur d'apparition d'un plancher de graviers limitant l'enracinement a été évaluée. Les facteurs du sol étaient fortement inter-dépendants. Les infections induites par C. fusipes diminuaient lorsque les traces d'hydromorphie apparaissaient plus profondément. Différentes espèces de Phytophthora ont été détectées dans deux des trois parcelles. Les Phytophthora étaient alors présents à la base de la majorité des chênes. La détection positive des Phytophthora était fortement liée à la saison de piégeage. L'état sanitaire des chênes diminuait avec la sévérité des infections induites par C. fusipes et la profondeur d'apparition des traces d'hydromorphie. Au contraire, aucun lien n'a pu être mis en évidence entre l'état sanitaire des chênes et la présence du Phytophthora dans le sol

Relationships between soil factors, Quercus robur health, Collybia fusipes root infection and Phytophthora presence

About sixty oaks (Quercus robur), with different status of crown health, located in three plots in north eastern France were assessed for their degree of infection by C. fusipes. Baiting of Phytophthora spp. from soil was carried out at different seasons in a sub sample of these oaks. The variability of soil characteristics, such as depth to waterlogging signs and depth to a gravel layer limiting rooting was evaluated. Soil factors were tightly correlated. Infection induced by C. fusipes was significantly more severe in non-waterlogged conditions. In two plots Phytophthora spp. were present at the base of the majority of trees investigated. Success of Phytophthora isolation was significantly linked with the season. The crown health status deteriorates with increasing severity of infection by C. fusipes and with the depth to waterlogging. Conversely, Phytophthora was not correlated with crown health status.Relations entre les facteurs du sol, l'état du houppier de Quercus robur, les infections racinaires induites par Collybia fusipes et la présence de Phytophthora. Le niveau d'infection induit par Collybia fusipes a été évalué sur environ 60 chênes pédonculés (Quercus robur) présentant différents états sanitaires, situés dans trois parcelles du nord-est de la France. Le piégeage des Phytophthora spp. présents dans le sol a été effectué sur un sous-échantillon de chênes à différentes saisons. La variabilité des facteurs du sol concernant la profondeur d'apparition des traces d'hydromorphie et la profondeur d'apparition d'un plancher de graviers limitant l'enracinement a été évaluée. Les facteurs du sol étaient fortement inter-dépendants. Les infections induites par C. fusipes diminuaient lorsque les traces d'hydromorphie apparaissaient plus profondément. Différentes espèces de Phytophthora ont été détectées dans deux des trois parcelles. Les Phytophthora étaient alors présents à la base de la majorité des chênes. La détection positive des Phytophthora était fortement liée à la saison de piégeage. L'état sanitaire des chênes diminuait avec la sévérité des infections induites par C. fusipes et la profondeur d'apparition des traces d'hydromorphie. Au contraire, aucun lien n'a pu être mis en évidence entre l'état sanitaire des chênes et la présence du Phytophthora dans le sol

Le récepteur P2X7, une nouvelle cible thérapeutique dans la maladie d’Alzheimer

International audienceNo abstract availabl

Selection of similar single domain antibodies from two immune VHH libraries obtained from two alpacas by using different selection methods.

International audienceThe two most used methods to select camelid single-domain antibody-fragments (VHHs) are: displaying their repertoires on the surface of filamentous bacteriophages (phage display) or linking them to ribosomes (ribosome display). In this study, we compared specific VHHs isolated from two different immune libraries coming from two different alpacas by using these two selection methods. Three anti-GFAP (glial fibrillary acidic protein) VHHs were derived from an immune library obtained by ribosome display after immunization of one alpaca with purified GFAP, a protein expressed by astroglial cells. In parallel, three other anti-GFAP VHHs were derived from an immune library by phage display after immunization of another alpaca with a human brain tissue extract containing GFAP. All the VHHs were closely related and one VHH was found to be strictly identical in both studies. This highlights the selection pressure exerted by the camelid immune system to shape the paratope of an antibody against a defined antigen