Rôle des peptides natriurétiques dans la régulation de l'homéostasie glucidique

Il est maintenant bien établi que les niveaux circulants de peptides natriurétiques (PN) sont diminués lors de l'obésité et qu'une faible quantité est associée avec le développement futur d'un diabète de type 2 (DT2). Cependant aucune étude n'a à ce jour démontré un lien causal et mécanistique entre l'activité biologique des PN et le développement du DT2. Le but de cette thèse est d'étudier le rôle des PN au niveau du tissu adipeux et du muscle squelettique dans un contexte d'obésité et de DT2. Nous avons étudié dans un premier volet le lien entre expression et signalisation du système PN dans le tissu adipeux humain et l'insulinosensibilité. Nous avons pu montrer que l'expression du NPRA dans le tissu adipeux humain corrèle négativement avec l'insulinosensibilité et positivement avec l'expression de gènes impliqués dans la régulation du métabolisme glucidique adipocytaire. L'expression du NPRA diminue avec le grade d'obésité, le prédiabète et le diabète de type 2. Nous avons également mis en évidence un nouveau rôle biologique des PN en montrant qu'ils stimulent spécifiquement le transport de glucose dans l'adipocyte humain. Le mécanisme moléculaire implique l'activation de la signalisation d'Akt de manière GMPc-dépendante. Nous avons dans un deuxième volet mis en évidence que l'expression protéique du NPRA musculaire humain corrèle positivement avec la sensibilité à l'insuline systémique et est très diminuée chez les individus obèses. L'expression du récepteur de clairance aux PN (NPRC) est quant à lui plus élevé dans les muscles de sujets DT2. Ces résultats sont retrouvés chez la souris obèse/diabétique. De plus, nous avons montré qu'une perfusion chronique de BNP chez ces souris améliore leur contrôle glycémique ainsi que leur sensibilité à l'insuline. L'amélioration du métabolisme glucidique chez la souris DT2 s'accompagne d'une diminution de l'accumulation d'espèces lipotoxiques et d'une augmentation de la capacité oxydative lipidique musculaire. Ces résultats sont également reproduits sur des cellules musculaires humaines où un traitement chronique aux PN protège partiellement de la lipotoxicité induite par le palmitate. En résumé, cette thèse révèle que l'expression et l'activation du NPRA au niveau des muscles squelettiques et des adipocytes humains déterminent en partie la sensibilité à l'insuline et jouent un rôle clé dans le maintien de l'homéostasie glucidique.Natriuretic peptides (NP) levels are reduced in obesity and predict the risk of type 2 diabetes (T2D). Since skeletal muscle was recently shown as a key target tissue of NP, we aimed to investigate adipose and muscle NP receptor (NPR) signaling in the context of obesity and T2D. So, we studied control of glucose metabolism in adipocyte. We firstly demonstrated that NPRA expression is positively correlated with de novo lipogenesis gene expression in human adipose tissue. Then, we showed acute NP treatment in human adipocyte increase glucose uptake and oxidation in a p38 MAPK dependent manner. In parallel to this work, we found that muscle NPRA correlated positively with whole-body insulin sensitivity in humans, and was strikingly down-regulated in obese subjects and recovered in response to diet-induced weight loss. In addition, muscle NP clearance receptor (NPRC) increased in individuals with impaired glucose tolerance and T2D. Similar results were found in obese diabetic mice. Although no acute effect of BNP on insulin sensitivity was observed in lean mice, chronic BNP infusion improved blood glucose control and insulin sensitivity in skeletal muscle of obese and diabetic mice. This occurred in parallel of a reduced lipotoxic pressure in skeletal muscle due to an up­regulation of lipid oxidative capacity. In addition, chronic NP treatment in human primary myotubes increased lipid oxidation and reduced palmitate-induced lipotoxicity. Collectively, our data show that activation of NPRA signaling in skeletal muscle is important for the maintenance of long-term insulin sensitivity by decreasing lipotoxic pressure in skeletal muscle and by increasing glucose utilization in adipocytes

The Student Aerospace Challenge: a european multidisciplinary contest and tertiary educational programme

Inspired by the first successful tests of a private manned spaceplane in 2004, the Student Aerospace Challenge was created in 2006 by the European Astronaut Club and its partners - Dassault Aviation, the European Space Agency, the International Astronautical Federation, Safran and Thales at the time - to allow European university students to explore some aspects of manned suborbital vehicles. Until 2020, the Challenge focused on a local reusable vehicle reaching Mach 3.5 and an altitude of 100 km. Since the 15th edition, to better respond to the evolution of the sector, a second vehicle is proposed: a hypersonic vehicle dedicated to point-to-point transportation taking, for example, less than two hours to travel from Barcelona to Tokyo. Each year, the Steering Committee defines several work packages corresponding to a large variety of study domains realistically related to this type of innovative vehicles like aerodynamic and flight control, structure, reusable propulsion, airworthiness, promotion, market analysis, legal frame & medicine. The introduction of a second vehicle having a quite different mission led the Committee to introduce dedicated topics. In addition, for the current edition, a new work package was proposed to cover potential applications of suborbital flights other than carrying passengers. In function of their background and interest, European University students have the opportunity to work, during several months, on a topic related to one of the work packages and to explore new solutions. Proposed projects should be technically realistic, economically viable and environmentally friendly. Reports and posters issued by student teams are evaluated by the Steering Committee some weeks before the “Suborbital Day”, a dedicated event organised like a mini-symposium, usually on-site where students present orally their projects and meet representatives of the different partners. The best-quoted projects are rewarded with prizes, among them, the ESA Grand Prize offering the winner team the unique opportunity to present their project in an appropriate European space-related event. To date, 216 teams and 998 University students coming from all over Europe already took part in the Student Aerospace Challenge, a motivating and ambitious multidisciplinary educational programme. Their participation allowed them to complement their knowledge, learn new skills and enlarge their network in the space secto

Cationic poly(amidoamine) promotes cytosolic delivery of bovine RNase A in melanoma cells, while maintaining its cellular toxicity

Ribonucleases are known to cleave ribonucleic acids, inducing cell death. RNase A, a member of the ribonuclease family, generally displayed poor in vitro activity. This has been attributed to factors such as low intracellular delivery. Poly(amidoamine)s have been used to promote the translocation of non-permeant proteins to the cytosol. Our objective was to demonstrate that poly(amidoamine)s could potentially promote the delivery of RNase A to selected cell line. Interactions of three cationic poly(amidoamine)s (P1, P2 and ISA1) with wild-type bovine RNase A were investigated using gel retardation assays, DLS and microcalorimetry. Although the polymers and the protein are essentially cationic at physiological pH, complexation between the PAAs and RNase A was observed. The high sensitivity differential scanning calorimetry (HSDSC) thermograms demonstrated that the thermal stability of the protein was reduced when complexed with ISA1 (Tmax decreased by 6.5 °C) but was not affected by P1 and P2. All the polymers displayed low cytotoxicity towards non-cancerous cells (IC50 > 3.5 mg mL?1). While RNase A alone was not toxic to mouse melanoma cells (B16F1), P1 was able to promote cytosolic delivery of biologically active RNase A, increasing cell death (IC50 = 0.09 mg mL?1)

Using Bayesian Programming for Multi-Sensor Data Fusion in Automotive Applications

International audienceA prerequisite to the design of future Advanced Driver Assistance Systems for cars is a sensing sytem providing all the information required for high-level driving assistance tasks. Carsense is a European project whose purpose is to develop such a new sensing system. It will combine different sensors (laser, radar and video) and will rely on the fusion of the information coming from these sensors in order to achieve better accuracy, robustness and an increase of the information content. This paper demonstrates the interest of using probabilistic reasoning techniques to address this challenging multi-sensor data fusion problem. The approach used is called Bayesian Programming. It is a general approach based on an implementation of the Bayesian theory. It was introduced first to design robot control programs but its scope of application is much broader and it can be used whenever one has to deal with problems involving uncertain or incomplete knowledge

Using Bayesian Programming for Multi-Sensor Multi-Target Tracking in Automotive Applications

International audienceA prerequisite to the design of future Advanced Driver Assistance Systems for cars is a sensing system providing all the information required for high-level driving assistance tasks. Carsense is a European project whose purpose is to develop such a new sensing system. It will combine different sensors (laser, radar and video) and will rely on the fusion of the information coming from these sensors in order to achieve better accuracy, robustness and an increase of the information content. This paper demonstrates the interest of using probabilistic reasoning techniques to address this challenging multi-sensor data fusion problem. The approach used is called Bayesian Programming. It is a general approach based on an implementation of the Bayesian theory. It was introduced first to design robot control programs but its scope of application is much broader and it can be used whenever one has to deal with problems involving uncertain or incomplete knowledge

Poly(amidoamine)s synthesis, characterisation and interaction with BSA

Cationic poly(amidoamine)s (PAAs) were synthesised and characterised by NMR and gel permeation chromatography. Their thermal properties were investigated using thermogravimetric analysis and differential scanning calorimetry. Although poly(amidoamine)s have been used as endosomolytic polymers for protein intracellular delivery, the interaction of the polymers with the proteins still need to be investigated. BSA was used as a model protein and complexation with the different poly(amidoamine) s was investigated using gel retardation assays, fluorescence spectroscopy and high sensitivity differential scanning calorimetry. Our results indicate that the thermal stability of BSA was affected upon interaction and complexation with the poly(amidoamine)s, however these interactions did not seem to modify the structure of the protein. Polymer flexibility seemed to favour polymer/protein complexation and promoted thermal stability

Loss of Aip1 reveals a role in maintaining the actin monomer pool and an in vivo oligomer assembly pathway

Enhanced polymerization of actin in latrunculin A–treated aip1Δ cells shows that filament assembly does not occur from monomeric actin alone in vivo

A new role for human dyskerin in vesicular trafficking

Dyskerin is an essential, conserved, multifunctional protein found in the nucleolus, whose loss of function causes the rare genetic diseases X-linked dyskeratosis congenita and Hoyeraal-Hreidarsson syndrome. To further investigate the wide range of dyskerin's biological roles, we set up stable cell lines able to trigger inducible protein knockdown and allow a detailed analysis of the cascade of events occurring within a short time frame. We report that dyskerin depletion quickly induces cytoskeleton remodeling and significant alterations in endocytic Ras-related protein Rab-5A/Rab11 trafficking. These effects arise in different cell lines well before the onset of telomere shortening, which is widely considered the main cause of dyskerin-related diseases. Given that vesicular trafficking affects many homeostatic and differentiative processes, these findings add novel insights into the molecular mechanisms underlining the pleiotropic manifestation of the dyskerin loss-of-function phenotype

Poly(amidoamine)-BSA conjugates synthesised by Michael addition reaction retained enzymatic activity

Polymer-protein conjugates are key to overcome some of the therapeutic protein limitations, including inefficient intracellular delivery. Poly(amidoamine)s are bioresponsive polyelectrolytes, which can form complexes with proteins and promote their delivery into the cytosol of cells. To investigate if conjugation would affect the activity of the protein, two poly(amidoamine)-BSA conjugates were synthesised using a “grafted to” method and Michael addition reaction. Following purification, the conjugates were characterised by electrophoresis, size exclusion chromatography (Mn(C1) = 140.7 kDa ; Mn(C2) = 218.6 kDa) and light scattering (Dh(C1) = 37.5 nm ; Dh(C2) = 75.1 nm). As a result of the conjugation with the cationic polymer, the conjugates had a positive zeta potential (?(C1) = +15.4 mV; ?(C2) = +20.2 mV). TNBS assays demonstrated that 16% to 25% of the protein amine groups were modified and HPLC analysis indicated that the amount of protein in the conjugate was 0.76 mg of BSA/mg of PAA (C1) and 0.43 mg of BSA /mg of PAA (C2). Enzymatic assays indicated the conjugates displayed an esterase activity similar (C1) or reduced ~ 35% (C2) compare to BSA. Altogether the results demonstrated that the conjugation of poly(amidoamine)s to a model protein can lead to the formation of bioconjugates that retain the enzymatic activity of the native protein. Such conjugates could have some application in protein delivery and enzyme engineering for biocatalysis and biosensors