Quantitative Proteomic Approach Reveals Altered Metabolic Pathways in Response to the Inhibition of Lysine Deacetylases in A549 Cells under Normoxia and Hypoxia.

Growing evidence is showing that acetylation plays an essential role in cancer, but studies on the impact of KDAC inhibition (KDACi) on the metabolic profile are still in their infancy. Here, we analyzed, by using an iTRAQ-based quantitative proteomics approach, the changes in the proteome of KRAS-mutated non-small cell lung cancer (NSCLC) A549 cells in response to trichostatin-A (TSA) and nicotinamide (NAM) under normoxia and hypoxia. Part of this response was further validated by molecular and biochemical analyses and correlated with the proliferation rates, apoptotic cell death, and activation of ROS scavenging mechanisms in opposition to the ROS production. Despite the differences among the KDAC inhibitors, up-regulation of glycolysis, TCA cycle, oxidative phosphorylation and fatty acid synthesis emerged as a common metabolic response underlying KDACi. We also observed that some of the KDACi effects at metabolic levels are enhanced under hypoxia. Furthermore, we used a drug repositioning machine learning approach to list candidate metabolic therapeutic agents for KRAS mutated NSCLC. Together, these results allow us to better understand the metabolic regulations underlying KDACi in NSCLC, taking into account the microenvironment of tumors related to hypoxia, and bring new insights for the future rational design of new therapies

Analyse comparée des écosystèmes digestifs du rumen de la vache et du caecum du lapin

Dans cette revue nous avons synthétisé les données obtenues dans notre équipe et celles de la bibliographie afin de contribuer à une meilleure connaissance de l’écologie des communautés bactériennes et archées des fermenteurs digestifs des mammifères herbivores. L’analyse a porté sur la comparaison des deux principales stratégies digestives rencontrées chez les mammifères herbivores actuels : un fermenteur en position proximale, le rumen, et un fermenteur en position distale, le caecum. Parmi les espèces d’intérêt agronomique,la vache et le lapin on été choisis comme animaux modèles. Après avoir rappelé les caractéristiques anatomiques et physicochimiques de ces fermenteurs digestifs, nous avons analysé les spécificités de leurs communautés procaryotiques liées à l’hôte, la variabilité individuelle, la structuration spatiale (inter- et intra- fermenteurs digestifs) et la dynamique temporelle (journalière et hebdomadaire) avec ou sans perturbation nutritionnelle induite

Comparison of chromosomal imbalances in neuroendocrine and non-small-cell lung carcinomas.

International audienceLung carcinomas are represented by non-small-cell lung carcinomas (NSCLC) and neuroendocrine carcinomas (NE) which differ in their clinical presentation and prognosis. We used comparative genomic hybridization (CGH) to characterize and compare the chromosomal pattern of 11 NSCLC and 11 high-grade NE lung carcinomas. Overall, the total number of aberrations was higher in NSCLC than in high-grade NE lung tumors (p < 0.05) and gains predominated over losses in NSCLC (p < 0.0003). Gains common to both lung tumor phenotypes were detected in 1p, 1q, 3q, 5p, 6p, 8q, 12, 17q, 19p, 19q, 20p, 20q, and X, whereas common losses were found in 2q, 3p, 4p, 4q, 5q, 8p, 9p, 10p, 11p, 11q, 13q, and 17p. Major gains on 18q and losses on 2p and 16q were exclusively detected in high-grade NE lung tumors. On the other hand, major gains on 2p and 15q and losses on 21q were found only in NSCLC. Furthermore, gains within 22q11-q12 and 7p12-p15 were associated with NSCLC (p < 0.05). The differences in the pattern and distribution of genetic changes observed in NSCLC as opposed to high-grade NE lung carcinomas suggest the existence of distinct tumorigenic pathways between these two major classes of lung tumors

[Secretome: definitions and biomedical interest]

International audienceINTRODUCTION: The secretome, or secretomics refers to the global study of proteins that are secreted by a cell, a tissue or an organism. EXEGESIS: The secretome is an important class of proteins that control many biological and physiological processes. Many secretion pathways are implicated in the release of these proteins. CONCLUSION: The secretome is a potential source suitable for the discovery of new therapeutic targets or biomarker candidates

Peptides OFFGEL electrophoresis: a suitable pre-analytical step for complex eukaryotic samples fractionation compatible with quantitative iTRAQ labeling.

International audienceABSTRACT: BACKGROUND: The proteomes of mammalian biological fluids, cells and tissues are complex and composed of proteins with a wide dynamic range. The effective way to overcome the complexity of these proteomes is to combine several fractionation steps. OFFGEL fractionation, recently developed by Agilent Technologies, provides the ability to pre-fractionate peptides into discrete liquid fractions and demonstrated high efficiency and repeatability necessary for the analysis of such complex proteomes. RESULTS: We evaluated OFFGEL fractionator technology to separate peptides from two complex proteomes, human secretome and human plasma, using a 24-wells device encompassing the pH range 3-10. In combination with reverse phase liquid chromatography, peptides from these two samples were separated and identified by MALDI TOF-TOF. The repartition profiles of the peptides in the different fractions were analyzed and explained by their content in charged amino acids using an algorithmic model based on the possible combinations of amino acids. We also demonstrated for the first time the compatibility of OFFGEL separation technology with the quantitative proteomic labeling technique iTRAQ allowing inclusion of this technique in complex samples comparative proteomic workflow. CONCLUSION: The reported data showed that OFFGEL system provides a highly valuable tool to fractionate peptides from complex eukaryotic proteomes (plasma and secretome) and is compatible with iTRAQ labeling quantitative studies. We therefore consider peptides OFFGEL fractionation as an effective addition to our strategy and an important system for quantitative proteomics studies

Adaptation rapide de la communauté bactérienne cæcale du lapin à une réduction du taux de fibres alimentaires

CT3 : Digestion, alimentation et valeur des alimentsNational audienceLes maladies digestives sont une des causes majeures de mortalité et de pertes économiques dans les élevages cunicoles. L’objectif de travail de notre équipe est de contribuer à l’émergence de solutions alternatives à l’antibiothérapie. Dans ce sens, nous avons déjà démontré qu’une augmentation du taux de fibres dans la ration améliorait la santé digestive du lapin. Cependant l’impact sur l’équilibre du microbiote cæcal n’est pas connu. Ainsi, cette étude a pour objectif d’étudier la réponse de l’écosystème cæcal du lapin à une perturbation nutritionnelle basée sur une réduction du taux de fibres

Cardiac phosphoproteome reveals cell signaling events involved in doxorubicin cardiotoxicity.

International audienceThe successful use of anthracyclines like doxorubicin in chemotherapy is limited by their severe cardiotoxicity. Despite decades of clinical application, a satisfying description of the molecular mechanisms involved and a preventive treatment have not yet been achieved. Here we address doxorubicin-induced changes in cell signaling as a novel potential mediator of doxorubicin toxicity by applying a non-biased screen of the cardiac phosphoproteome. Two-dimensional gel electrophoresis, phosphospecific staining, quantitative image analysis, and MALDI-TOF/TOF mass spectrometry were combined to identify (de)phosphorylation events occurring in the isolated rat heart upon Langendorff-perfusion with clinically relevant (5μM) and supraclinical concentrations (25μM) of doxorubicin. This approach identified 22 proteins with a significantly changed phosphorylation status and these results were validated by immunoblotting for selected phosphosites. Overrepresentation of mitochondrial proteins (>40%) identified this compartment as a prime target of doxorubicin. Identified proteins were mainly involved in energy metabolism (e.g. pyruvate dehydrogenase and acyl-CoA dehydrogenase), sarcomere structure and function (e.g. desmin) or chaperone-like activities (e.g. α-crystallin B chain and prohibitin). Changes in phosphorylation of pyruvate dehydrogenase, regulating pyruvate entry into the Krebs cycle, and desmin, maintaining myofibrillar array, are relevant for main symptoms of cardiac dysfunction related to doxorubicin treatment, namely energy imbalance and myofibrillar disorganization. This article is part of a Special Issue entitled: Translational Proteomics SI: Translational Proteomics

Caractérisation comparée des communautés bactériennes du contenu cæcal, des cæcotrophes et des fæces dures chez le lapin par PCR-SSCP

National audienc

Characterization of bacterial communities in caecum, hard and soft feces of rabbit using 16S rRNA genes capillary electrophoresis single-strand conformation polymorphism (CE-SSCP).

International audienceDynamic and individual variability of caecal bacterial community were compared to those of hard and soft feces using capillary electrophoresis single strand conformation polymorphism (CE-SSCP). Soft and hard feces of 14 adult rabbits were weekly sampled during 5 weeks while caecal content was sampled on the 3rd (after surgery) and 5th week (after sacrifice). Bacterial communities were compared for their structure (peaks of CE-SSCP profiles analysis) and their diversity (estimation of Simpson diversity index on CE-SSCP profiles, D’). Bacterial community of caecal content, soft and hard feces presented individual variability (P<0.05 for diversity and structure). Without disturbance (1st to 3rd and 4th to 5th weeks), bacterial communities of soft and hard feces remained constant in time for diversity and structure. Sampling caecal content by surgical way greatly modified all bacterial communities (P<0.05 for diversity and structure) suggesting to find an alternative sample type to realize dynamic studies of bacterial caecal community within a same individual. The bacterial communities of the 3 types of sample were different (P<0.05), but presented a similar diversity (D’=3.96±0.48; NS). However, bacterial community of caecal content was closer to that of soft feces than that of hard feces (R of ANOSIM=0.13 and 0.24, respectively) and differed only on some peaks. Therefore, we can assume that soft feces could be used to realize dynamic studies of caecal bacterial community