Résumé : Les biofilms électroactifs (EA) sont des consortia bactériens mono ou pluri-espèces, qui ont la faculté d’échanger des électrons provenant de leur métabolisme avec les surfaces solides conductrices des électrodes. Cette découverte est à l’origine d’un nouveau dispositif énergétique : la pile à combustible microbienne (PACM). Depuis les années 2000, la littérature scientifique sur les biofilms EA et sur les PACM explose, notamment grâce à la découverte de bactéries capables de transférer par voie directe – par des pili ou protéines transmembranaires – des électrons vers les électrodes. Plusieurs sources de bactéries EA ont été à ce jour découvertes, allant de cultures pures à des communautés bactériennes plus complexes. Ces dernières sont issues de milieux aqueux naturels (milieux marins ou d’eau douce), industriels ou urbains (effluents d’industrie, eaux usées domestiques). La plupart de ces sources bactériennes proviennent d’environnements de climat tempéré. Dans ce travail de thèse, plusieurs sols de milieux humides de la Guyane ont été identifiés comme étant de sources de bactéries EA. Les expériences menées sous potentiel d’électrode imposé et constant ont permis d’étudier l’adhésion à la surface d’électrode des biofilms EA issus de la flore endogène des milieux sélectionnés. La formation de bioanodes et de biocathodes a été possible en présence respectivement d’acétate et d’oxygène dans les milieux. Une étude par voltammétrie cyclique a mis en évidence les pics d’oxydo-réduction en lien avec les échanges électroniques du biofilm EA et de l’électrode. En optimisant la procédure de formation des biofilms EA par chronoampérométrie (surface d’électrode plus importante, apport en continu et progressif du substrat), une densité de courant maximale de 12 A/m2 et un rendement faradique de 24 % ont été obtenus. Une autre méthode pour former des biofilms EA à partir d’un milieu choisi, la mangrove, a consisté à utiliser deux prototypes de PACM : une pile à compartiment unique et à cathode à air, et une pile benthique. Dans les deux cas, les biofilms EA ont pu être formés et étudiés, complétant certaines observations faites sous potentiel imposé. La PACM benthique s’est avérée être la plus proche d’une application à grande échelle puisqu’elle a été complètement autonome : anode et cathode utilisant uniquement le milieu pour fonctionner. Elle a pu être étudiée en laboratoire comme sur le terrain.
Electroactive biofilms (EA) are mono or multi-species bacterial consortia, which have the ability to exchange electrons from their metabolism with solid surfaces of conductive electrodes. This discovery is the basis for a device of energy production: microbial fuel cell (MFC). Since the 2000s, the scientific literature on EA biofilms and MFC explodes, thanks to the discovery of bacteria that are able to transfer directly – by pili or trans-membrane proteins – electrons to electrodes. Several sources of EA bacteria were discovered to date, ranging from pure cultures to more complex bacterial communities. Those last are from natural (marine and freshwater), industrial or urban (industrial effluents, domestic wastewater) aqueous environments. However, the vast majority of these are from temperate environments.In this thesis, several wetland soils of French Guiana have been identified as sources of EA bacteria. Experiments under poised and constant electrode potential were used to examine adherence to the electrode surface of EA biofilms from the endogenous flora of selected environments. Formation of bioanodes and biocathodes was possible in the presence respectively of acetate and oxygen in the media. A study by cyclic voltammetry showed the redox peaks related to electronic exchanges between EA biofilm and electrode. By optimizing the process of EA biofilm formation by chronoamperometry (larger electrode surface, providing continuous and progressive substrate), a maximum current density of 12 A/m2 and a coulombic efficiency of 24% were obtained.Another method to form EA biofilms from a chosen medium (mangrove) was to use two MFC prototypes: a single compartment and air cathode one, and a benthic one. In both cases, the EA biofilms have been trained and studied supplementing certain observations made under poised polarization. MFC benthic proved to be the closest to a wide application since it was completely autonomous, anode and cathode only using the medium to function. It has been studied in the laboratory and in the field.

http://www.theses.fr/2012AGUY0560/document

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Monocytes/macrophages (MΦ), considered as plastic cells, can differentiate into either a pro-inflammatory (M1) subtype, also known as a classically activated subtype, or an anti-inflammatory alternatively activated subtype (M2) according to their microenvironment. Phenotypic markers of mouse polarized MΦ have been extensively studied, whereas their human counterparts remain less characterized. The main goal of this study was therefore to carefully characterize phenotypic and genomic markers of primary human MΦ generated from M-CSF-treated blood monocytes and polarized towards M1 or M2 subtype upon the action of lipopolysaccharide and interferon-γ (for M1) or interleukin (IL)-4 (for M2). Membrane expression of the markers CD80 and CD200R was found to be specific of human M1 and M2 polarized MΦ, respectively, whereas, by contrast, mannose receptor (CD206) expression did not discriminate between M1 and M2. mRNA expression analysis further identified six markers of M1 polarization (IL-12p35, CXCL10, CXCL11, CCL5, CCR7 and IDO1), five markers of M2 polarization (TGF-β, CCL14, CCL22, SR-B1 and PPARγ) and transcription factors involved in MΦ polarization. Ability of human M-CSF-generated MΦ to polarize toward M1 or M2 subtype was also associated with enhanced secretion of TNFα, IL-1β, IL-12p40, CXCL10 and IL-10 (for M1) or CCL22 (for M2). Moreover, the comparison of the expression of M1 markers in M-CSF- and GM-CSF-MΦ polarized towards M1 subtype has revealed similarities. In conclusion, we demonstrated that human M-CSF MΦ can polarize toward a M1 type after IFNγ/LPS stimulation. Moreover, the M1 and M2 markers of human polarized MΦ identified in the present study may be useful to better identify human MΦ subtypes, particularly at the tissue level, in order to better understand their respective roles in the development of pathologies.
ISSN: 0008-8749

inserm-00823273
http://www.hal.inserm.fr/inserm-00823273
DOI : 10.1016/j.cellimm.2013.01.010
PUBMED : 23454681

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Human hepatocytes cultured in a monolayer configuration represent a well-established in vitro model in liver toxicology, notably used in drug transporter studies. Polarized status of drug transporters, i.e., their coordinated location at sinusoidal or canalicular membranes, remains however incompletely documented in these cultured hepatocytes. The present study was therefore designed to analyze transporter expression and location in such cells. Most of drug transporters were first shown to be present at notable mRNA levels in monolayer-cultured human hepatocytes. Cultured human hepatocytes, which morphologically exhibited bile canaliculi-like structures, were next demonstrated, through immunofluorescence staining, to express the influx transporters organic anion transporting polypeptide (OATP) 1B1, OATP2B1 and organic cation transporter (OCT) 1 and the efflux transporter multidrug resistance-associated protein (MRP) 3 at their sinusoidal pole. In addition, the efflux transporters P-glycoprotein and MRP2 were detected at the canalicular pole of monolayer-cultured human hepatocytes. Moreover, canalicular secretion of reference substrates for the efflux transporters bile salt export pump, MRP2 and P-glycoprotein as well as sinusoidal drug transporter activities were observed. This polarized and functional expression of drug transporters in monolayer-cultured human hepatocytes highlights the interest of using this human in vitro cell model in xenobiotic transport studies.
ISSN: 0887-2333

hal-01141725
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01141725
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01141725/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01141725/file/Polarized%20location%20of%20SLC.pdf
DOI : 10.1016/j.tiv.2015.03.019

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
The outer membrane proteins (OMPs) of Gram-negative bacteria play a crucial role in virulence and pathogenesis. Identification of these proteins represents an important goal for bacterial proteomics, because it aids in vaccine development. Here, we have developed such an approach for Ehrlichia ruminantium, the obligate intracellular bacterium that causes heartwater. A preliminary whole proteome analysis of elementary bodies, the extracellular infectious form of the bacterium, had been performed previously, but information is limited about OMPs in this organism and about their role in the protective immune response. Identification of OMPs is also essential for understanding Ehrlichia’s OM architecture, and how the bacterium interacts with the host cell environment. First, we developed an OMP extraction method using the ionic detergent sarkosyl, which enriched the OM fraction. Second, proteins were separated via one-dimensional electrophoresis, and digested peptides were analyzed via nano-liquid chromatographic separation coupled with mass spectrometry (LCMALDI-TOF/TOF). Of 46 unique proteins identified in the OM fraction, 18 (39%) were OMPs, including 8 proteins involved in cell structure and biogenesis, 4 in transport/virulence, 1 porin, and 5 proteins of unknown function. These experimental data were compared to the predicted subcellular localization of the entire E. ruminantium proteome, using three different algorithms. This work represents the most complete proteome characterization of the OM fraction in Ehrlichia spp. The study indicates that suitable subcellular fractionation experiments combined with straightforward computational analysis approaches are powerful for determining the predominant subcellular localization of the experimentally observed proteins.We identified proteins potentially involved in E. ruminantium athogenesis, which are good novel targets for candidate vaccines. Thus, combining bioinformatics and proteomics, we discovered new OMPs for E. ruminantium that are valuable data for those investigating new vaccines against this organism. In summary, we provide both pioneering data and novel insights into the pathogenesis of this obligate intracellular bacterium.
ISSN: 1932-6203

hal-01203402
https://hal.univ-antilles.fr/hal-01203402
https://hal.univ-antilles.fr/hal-01203402/document
https://hal.univ-antilles.fr/hal-01203402/file/2015_Moumene_PLOSONE-1.pdf
DOI : 10.1371/journal.pone.0116758

Éditeur(s) : HAL CCSD Springer Verlag (Germany)
Résumé : International audience
Tight control of basal cytosolic Ca2+ concentration is essential for cell survival and to fine-tune Ca2+-dependent cell functions. A way to control this basal cytosolic Ca2+ concentration is to regulate membrane Ca2+ channels including store-operated Ca2+ channels and secondary messenger-operated channels linked to G-protein-coupled or tyrosine kinase receptor activation. Orai, with or without its reticular STIM partner and Transient Receptor Potential (TRP) proteins, were considered to be the main Ca2+ channels involved. It is well accepted that, in response to cell stimulation, opening of these Ca2+ channels contributes to Ca2+ entry and the transient increase in cytosolic Ca2+ concentration involved in intracellular signaling. However, in various experimental conditions, Ca2+ entry and/or Ca2+ currents can be recorded at rest, without application of any experimental stimulation. This led to the proposition that some plasma membrane Ca2+ channels are already open/activated in basal condition, contributing therefore to constitutive Ca2+ entry. This article focuses on direct and indirect observations supporting constitutive activity of channels belonging to the Orai and TRP families and on the mechanisms underlying their basal/constitutive activities.
ISSN: 0175-7571

inserm-01526286
http://www.hal.inserm.fr/inserm-01526286
http://www.hal.inserm.fr/inserm-01526286/document
http://www.hal.inserm.fr/inserm-01526286/file/EBJO-D-16-00056_R1-2%20%282%29.pdf
PUBMED : 28516266

Éditeur(s) : HAL CCSD Federation of American Society of Experimental Biology
Résumé : International audience
The innate immune system is able to detect bacterial LPS through the pattern recognition receptor CD14, which delivers LPS to various TLR signaling complexes that subsequently induce intracellular proinflammatory signaling cascades. In a previous study, we showed the overproduction of the soluble form of CD14 (sCD14) by macrophages from patients with cystic fibrosis (CF). CF is an autosomal recessive disorder that is caused by mutations in the gene that encodes the CFTR protein and is characterized by persistent inflammation. Macrophages play a significant role in the initial stages of this disease due to their inability to act as suppressor cells, leading to chronic inflammation in CF. In this work, we investigated the origin of sCD14 by human macrophages and studied the effect of sCD14 on the production of inflammatory cytokine/chemokine. Our data indicate that sCD14 stimulate proinflammatory cytokine/chemokine production in a manner that is independent of LPS but dependent on the TLR-4/CD14 membrane complex, NF-κB, and the inflammasome. Therefore, sCD14, overproduced by CF macrophages, originates primarily from the endocytosis/exocytosis process and should be considered to be a danger-associated molecular pattern. This elucidation of the origin and inflammation-induced mechanisms associated with sCD14 contributes to our understanding of maintained tissue inflammation.
ISSN: 0892-6638

hal-01526434
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01526434
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01526434/document
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01526434/file/Soluble%20CD14%20acts%20as%20a%20DAMP%20in%20human-accepted.pdf
PUBMED : 28122919

Éditeur(s) : HAL CCSD Springer Verlag (Germany) Springer Verlag
Résumé : Dihydroceramide ?4-desaturase 1 (DES1) catalyzes the last step of the de novo ceramide biosynthesis, which consists of the introduction of a trans ?4-double bond in the carbon chain of the dihydroceramide. It was previously observed that myristic acid binds DES1 through N-myristoylation. This N-terminal modification significantly increased the activity of the recombinant DES1 in COS-7 cells and targeted part of the enzyme initially present in the endoplasmic reticulum to the mitochondrial outer membrane, leading to an increase in ceramide levels. Since these results were obtained in a recombinant COS-7 cell model with high expression of rat DES1, the purpose of the present study was to investigate if the native DES1 enzyme was really upregulated by its N-myristoylation in cultured rat hepatocytes. We first showed that DES1 was the main dihydroceramide desaturase isoform expressed in rat hepatocytes. In this model, the wild-type myristoylable recombinant form of rat DES1 was found in both the endoplasmic reticulum and the mitochondria whereas the mutated non-myristoylable recombinant form (N-terminal glycine replaced by an alanine) was almost exclusively localized in the endoplasmic reticulum, which evidenced the importance of the myristoylation. Then, we showed that compared to other fatty acids, myristic acid was the only one to increase native DES1 activity, in both total cell lysates and mitochondrial fractions. The myristic acid-associated increase in DES1 activity was not linked to elevated mRNA or protein expression but more likely to its N-terminal myristoylation. Finally, the myristic acid-associated increase in DES1 activity slightly enhanced the number of apoptotic cells.
ISSN: 0024-4201

hal-00808026
https://hal-agrocampus-ouest.archives-ouvertes.fr/hal-00808026
DOI : 10.1007/s11745-011-3638-x

Éditeur(s) : HAL CCSD
Résumé : The ecological success of cnidarian-dinoflagellate symbiosis mainly relies on nutrient recycling. Environmental changes, such as global warming or pollution, often result to symbiosis breakdown, also called cnidarian bleaching. The understanding of mechanisms regulating the symbiosis establishment, maintenance and breakdown is essential to prevent massive bleaching phenomena. In this respect, my PhD project focused on the characterization of chemical mediators expressed in the sea anemone Anemonia viridis, their localization and their modulation by stress conditions. A comparative study was first conducted to characterize the chemical mediators and analyze their distribution within the symbiotic sea anemone. We described a great abundance and diversity of lipids in A. viridis tissues. From these results, we proposed possible transfers of FAs between the symbiotic partners. A thermal stress and a chemical stress have also been applied in laboratory-controlled conditions in order to induce symbiosis breakdown and bleaching of the sea anemones, in order to correlate A. viridis metabolome to its symbiotic status. A mapping of these metabolites has been performed by MALDI-MSI of tentacle cross-sections, as well as their evolution following stress. Some betaine lipids have thus been proposed as short-term indicators of stress. A. viridis stress response has also been evaluated with a lipidomic approach, and allowed to identify 3 lipid indicators of early stress response based on membrane fluidity markers. Overall, this study provides insight on key chemical mediators that may regulate the symbiosis maintenance, and may contribute to the symbiosis breakdown.
Le succès évolutif des Cnidaires symbiotiques réside en grande partie dans leurs échanges trophiques établis avec les Dinoflagellés du genre Symbiodinium. Cependant, le réchauffement climatique global ainsi que les pollutions ont un impact fort sur les écosystèmes coralliens, notamment en conduisant à la rupture de la symbiose, phénomène appelé blanchissement. La compréhension des mécanismes qui régissent l’établissement, le maintien et la rupture de la symbiose est essentielle à la prévention des épisodes de blanchissement massif. Dans ce contexte, les objectifs de mon projet de thèse sont de caractériser les médiateurs chimiques de l’anémone de mer Anemonia viridis, de les localiser, et d’analyser leur réponse face à un stress. Parmi les composés caractérisés, les lipides et les bétaïnes sont les plus abondants et présentent une grande diversité. Certains sont transférés des symbiotes vers l’hôte. Des anémones de mer ont ensuite été traitées en laboratoire afin de provoquer la rupture de la symbiose et le blanchissement des individus. Une étude cinétique a été menée par une approche globale comparative identique à celle réalisée sur l’anémone symbiotique. Par ailleurs, une cartographie de l’évolution de composés clé a été réalisée par MALDI-MSI. La réponse au stress a été évaluée et a permis d’identifier des lipides de bétaïne et trois indicateurs lipidiques comme marqueurs de réponse précoce au stress. L’ensemble de ces résultats apporte de nouveaux éléments de réponse concernant le rôle des médiateurs chimiques clés dans le maintien de la symbiose, ainsi que leur influence sur sa rupture.
https://tel.archives-ouvertes.fr/tel-01310668

NNT : 2015NICE4108
tel-01310668
https://tel.archives-ouvertes.fr/tel-01310668
https://tel.archives-ouvertes.fr/tel-01310668/document
https://tel.archives-ouvertes.fr/tel-01310668/file/2015NICE4108.pdf

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
Pre-association of CD95 at the plasma membrane is mandatory for efficient death receptor signaling. This homotrimerization occurs through self-association of an extracellular domain called the pre-ligand assembly domain (PLAD). Using novel molecular and cellular tools, we confirmed that CD95-PLAD is necessary to promote CD95 multimerization and plays a pivotal role in the transmission of apoptotic signals. However, while a human CD95 mutant deleted of the previously described PLAD domain (amino acids 1 to 66) fails to interact with its wild-type counterpart and trigger autonomous cell death, deletion of amino acids 1 to 42 does not prevent homo- or hetero (human/mouse)-oligomerization of CD95, and thus does not alter transmission of the apoptotic signal. Overall, these findings indicate that the region between amino acids 43 to 66 corresponds to the minimal motif involved in CD95 homotypic interaction and is necessary to convey an efficient apoptotic signal. Interfering with this PLAD may represent a new therapeutic strategy for altering CD95-induced apoptotic and non-apoptotic signals.
ISSN: 1932-6203

hal-00873708
https://hal.archives-ouvertes.fr/hal-00873708
https://hal.archives-ouvertes.fr/hal-00873708/document
https://hal.archives-ouvertes.fr/hal-00873708/file/Precise_Mappingsur the_CD95-accepted.pdf
DOI : 10.1371/journal.pone.0046236

Éditeur(s) : HAL CCSD National Academy of Sciences
Résumé : International audience
We applied single-molecule photobleaching to investigate the stoichiometry of human Orai1 and Orai3 channels tagged with eGFP and expressed in mammalian cells. Orai1 was detected predominantly as dimers under resting conditions and as tetramers when coexpressed with C-STIM1 to activate Ca(2+) influx. Orai1 was also found to be tetrameric when coexpressed with STIM1 and evaluated following fixation. We show that fixation rapidly causes release of Ca(2+), redistribution of STIM1 to the plasma membrane, and STIM1/Orai1 puncta formation, and may cause the channel to be in the activated state. Consistent with this possibility, Orai1 was found predominantly as a dimer when coexpressed with STIM1 in living cells under resting conditions. We further show that Orai3, like Orai1, is dimeric under resting conditions and is predominantly tetrameric when activated by C-STIM1. Interestingly, a dimeric Orai3 stoichiometry was found both before and during application of 2-aminoethyldiphenyl borate (2-APB) to activate a nonselective cation conductance in its STIM1-independent mode. We conclude that the human Orai1 and Orai3 channels undergo a dimer-to-tetramer transition to form a Ca(2+)-selective pore during store-operated activation and that Orai3 forms a dimeric nonselective cation pore upon activation by 2-APB.
ISSN: 0027-8424

hal-00873833
https://hal.archives-ouvertes.fr/hal-00873833
DOI : 10.1073/pnas.1114814108
PUBMED : 21987805

Éditeur(s) : HAL CCSD Lippincott, Williams & Wilkins
Résumé : PURPOSE OF REVIEW: Malaria represents one of the most important selective factors affecting human populations. Several inherited diseases of red blood cells lead to resistance at the erythrocytic stage. Among patients who experience hereditary elliptocytosis related to mutations of erythrocyte membrane proteins, molecular studies have shown the prevalence of particular spectrin mutations in patients from black ethnic extraction, leading one to question the selection of new malaria-resistant genes. RECENT FINDINGS: Prospective epidemiological and molecular studies in West Africa have confirmed the prevalence (between 0.6 and 1.6%) of particular spectrin mutations related to hereditary elliptocytosis. These studies have also revealed the frequency of alpha-spectrin chain polymorphisms, associated in cis with elliptocytogenic spectrin mutations and defining particular spectrin allele haplotypes. Culture studies of Plasmodium falciparum in elliptocytes bearing such elliptocytogenic alleles of spectrin showed that these alleles are supplementary genetic factors of malaria resistance in vitro. SUMMARY: Certain instances of spectrin mutations or polymorphisms have not yet been shown to constitute new factors of innate resistance to malaria in vivo. Epidemiological surveys of hereditary elliptocytosis and parasite culture studies, however, have argued that the relationships between parasite and spectrin-based skeleton should be examined more closely and the molecular interactions between parasite ligands and particular spectrin chain domains should be characterized.
ISSN: 1065-6251

inserm-00128119
http://www.hal.inserm.fr/inserm-00128119
http://www.hal.inserm.fr/inserm-00128119/document
http://www.hal.inserm.fr/inserm-00128119/file/Dhermy_et_al_figures.pdf
DOI : 10.1097/MOH.0b013e3280d21afd

Éditeur(s) : HAL CCSD Station Biologique
Résumé : International audience
Four "morphotypes" of Mytilidae were collected from wood falls in the west Pacific (Vanuatu islands) and in the Bohol sea (Philippines) between 300 and 1800 metre depth. Results from our study demonstrated the existence of extracellular bacteria located between microvilli at the apical surface of the cells all along the lateral zone of the gill filaments in the four mytilid morphotypes analyzed. Based on TEM observations, these Gram-negative bacteria were not methanotrophic due to the lack of concentric stacking of intracellular membranes in their cytoplasm. Based on FISH experiments, these bacteria belong to g-Proteobacteria. These preliminary observations described gill symbiosis-like interactions in metazoan species associated with sunken wood, strengthening the hypothesis that decomposing wood may serve as steps for the introduction of symbiotic Mytilidae to vents and seeps. Phylogenetic analysis of these extracellular bacteria are in progress in our lab in order to compare their 16SrDNA sequences to that of other marine invertebrate symbionts described to date.
ISSN: 0007-9723

hal-00753023
https://hal.univ-antilles.fr/hal-00753023

Éditeur(s) : HAL CCSD
Résumé : Communication between tumoral cells and the microenvironnement plays an essential role in the developpement of tumors. In that context, the new family of metalloproteases, the ADAM proteins, are major regulators of the tumoral progression by acting on the bioavaibility of importants mediators of cellular communication as cytokines and growth factors. Among this family, ADAM12 is the most associated with cancer. It has been shown to mediate signaling pathways by a process independant of its metalloproteasis activity, in particular for TGFβ signaling. This study show for the first time a role for the membrane form of ADAM12 in the induction of epithelial to mesenchymal transition (EMT), a essential process involved in tumor invasion, whom TGFβ is a main inducer. This effect is mediated by the activation of TGFβ signaling pathways, SMAD3, AKT and ERK and require the cytoplasmic tail of ADAM12L but not its catalytic activity. Activation of these pathways could involve a relocalisation of ADAM12L in special signaling platform in lipid rafts. Moreover, we have shown that ADAM12L interact with ZO-1 and ZO-2, two proteins of tight junctions, and could facilitate their desassembling during EMT. This work underscore for the first time a new function of ADAM12L in EMT, by a mecanism invovlving a modulation of signals regulating this process. A better understanding of the dynamic of these molecular mecanisms could allow the developpement of new targeted therapies to fight against tumoral progression.
Les échanges entre les cellules tumorales et le microenvironnement jouent un rôle essentiel dans le développement des tumeurs. Dans ce contexte, la nouvelle famille de métalloprotéases, les protéines ADAM, constituent aujourd’hui des régulateurs majeurs de la progression tumorale en agissant sur la biodisponibilité des médiateurs de la communication cellulaire que sont les cytokines, chimiokines et facteurs de croissance. Au sein de cette famille, ADAM12 est la plus associée au cancer. Elle possède la particularité de jouer un rôle dans la signalisation cellulaire, de façon indépendante de son activité métalloprotéase, notamment dans les voies de signalisation du TGFβ. Notre étude montre pour la première fois un rôle pour la forme membranaire d'ADAM12 dans l'induction de la transition épithélio-mésenchymateuse (EMT), un processus essentiel à l'invasion tumorale dont le TGFβ est un inducteur majeur. Cet effet est médié par l'activation des voies de signalisation du TGFβ, impliquant les protéines SMAD3, AKT et ERK et requiert le domaine cytoplasmique d'ADAM12L mais pas son domaine catalytique. L'activation de ces voies de signalisation pourrait impliquer une relocalisation d'ADAM12L au sein de plates-formes de signalisation dans les radeaux lipidiques. Par ailleurs, nous avons montré qu'ADAM12L interagit avec les protéines ZO-1 et ZO-2, des protéines des jonctions serrées, et pourrait favoriser leur désassemblage au cours de l'EMT. Nos travaux ont permis de mettre en évidence une nouvelle fonction pour ADAM12L dans l'EMT, par un mécanisme impliquant une modulation des signaux régulant ce processus. Une meilleure compréhension de la dynamique de ces mécanismes moléculaires pourrait permettre de développer de nouvelles thérapies ciblées pour lutter contre la progression tumorale.
https://tel.archives-ouvertes.fr/tel-01267577

Droits : info:eu-repo/semantics/OpenAccess
NNT : 2015REN1S063
tel-01267577
https://tel.archives-ouvertes.fr/tel-01267577
https://tel.archives-ouvertes.fr/tel-01267577/document
https://tel.archives-ouvertes.fr/tel-01267577/file/RUFF_Michael.pdf

Éditeur(s) : HAL CCSD Wiley Open Access
Résumé : International audience
Estetrol (E4) is a natural estrogen with a long half-life produced only by the human fetal liver during pregnancy. The crystal structures of the estrogen receptor α (ERα) ligand-binding domain bound to 17β-estradiol (E2) and E4 are very similar, as well as their capacity to activate the two activation functions AF-1 and AF-2 and to recruit the coactivator SRC3. In vivo administration of high doses of E4 stimulated uterine gene expression, epithelial proliferation, and prevented atheroma, three recognized nuclear ERα actions. However, E4 failed to promote endothelial NO synthase activation and acceleration of endothelial healing, two processes clearly dependent on membrane-initiated steroid signaling (MISS). Furthermore, E4 antagonized E2 MISS-dependent effects in endothelium but also in MCF-7 breast cancer cell line. This profile of ERα activation by E4, uncoupling nuclear and membrane activation, characterizes E4 as a selective ER modulator which could have medical applications that should now be considered further.
ISSN: 1757-4676

hal-01118028
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01118028
PUBMED : 25214462
PUBMEDCENTRAL : PMC4287935
DOI : 10.15252/emmm.201404112