Identification of Keratinocyte Growth Factor as a Target of microRNA-155 in Lung Fibroblasts: Implication in Epithelial-Mesenchymal Interactions

International audienceBACKGROUND: Epithelial-mesenchymal interactions are critical in regulating many aspects of vertebrate embryo development, and for the maintenance of homeostatic equilibrium in adult tissues. The interactions between epithelium and mesenchyme are believed to be mediated by paracrine signals such as cytokines and extracellular matrix components secreted from fibroblasts that affect adjacent epithelia. In this study, we sought to identify the repertoire of microRNAs (miRNAs) in normal lung human fibroblasts and their potential regulation by the cytokines TNF-alpha, IL-1beta and TGF-beta. METHODOLOGY/PRINCIPAL FINDINGS: MiR-155 was significantly induced by inflammatory cytokines TNF-alpha and IL-1beta while it was down-regulated by TGF-beta. Ectopic expression of miR-155 in human fibroblasts induced modulation of a large set of genes related to "cell to cell signalling", "cell morphology" and "cellular movement". This was consistent with an induction of caspase-3 activity and with an increase in cell migration in fibroblasts tranfected with miR-155. Using different miRNA bioinformatic target prediction tools, we found a specific enrichment for miR-155 predicted targets among the population of down-regulated transcripts. Among fibroblast-selective targets, one interesting hit was keratinocyte growth factor (KGF, FGF-7), a member of the fibroblast growth factor (FGF) family, which owns two potential binding sites for miR-155 in its 3'-UTR. Luciferase assays experimentally validated that miR-155 can efficiently target KGF 3'-UTR. Site-directed mutagenesis revealed that only one out of the 2 potential sites was truly functional. Functional in vitro assays experimentally validated that miR-155 can efficiently target KGF 3'-UTR. Furthermore, in vivo experiments using a mouse model of lung fibrosis showed that miR-155 expression level was correlated with the degree of lung fibrosis. CONCLUSIONS/SIGNIFICANCE: Our results strongly suggest a physiological function of miR-155 in lung fibroblasts. Altogether, this study implicates this miRNA in the regulation by mesenchymal cells of surrounding lung epithelium, making it a potential key player during tissue injury

miR-199a-5p Is Upregulated during Fibrogenic Response to Tissue Injury and Mediates TGFbeta-Induced Lung Fibroblast Activation by Targeting Caveolin-1

As miRNAs are associated with normal cellular processes, deregulation of miRNAs is thought to play a causative role in many complex diseases. Nevertheless, the precise contribution of miRNAs in fibrotic lung diseases, especially the idiopathic form (IPF), remains poorly understood. Given the poor response rate of IPF patients to current therapy, new insights into the pathogenic mechanisms controlling lung fibroblasts activation, the key cell type driving the fibrogenic process, are essential to develop new therapeutic strategies for this devastating disease. To identify miRNAs with potential roles in lung fibrogenesis, we performed a genome-wide assessment of miRNA expression in lungs from two different mouse strains known for their distinct susceptibility to develop lung fibrosis after bleomycin exposure. This led to the identification of miR-199a-5p as the best miRNA candidate associated with bleomycin response. Importantly, miR-199a-5p pulmonary expression was also significantly increased in IPF patients (94 IPF versus 83 controls). In particular, levels of miR-199a-5p were selectively increased in myofibroblasts from injured mouse lungs and fibroblastic foci, a histologic feature associated with IPF. Therefore, miR-199a-5p profibrotic effects were further investigated in cultured lung fibroblasts: miR-199a-5p expression was induced upon TGFβ exposure, and ectopic expression of miR-199a-5p was sufficient to promote the pathogenic activation of pulmonary fibroblasts including proliferation, migration, invasion, and differentiation into myofibroblasts. In addition, we demonstrated that miR-199a-5p is a key effector of TGFβ signaling in lung fibroblasts by regulating CAV1, a critical mediator of pulmonary fibrosis. Remarkably, aberrant expression of miR-199a-5p was also found in unilateral ureteral obstruction mouse model of kidney fibrosis, as well as in both bile duct ligation and CCl4-induced mouse models of liver fibrosis, suggesting that dysregulation of miR-199a-5p represents a general mechanism contributing to the fibrotic process. MiR-199a-5p thus behaves as a major regulator of tissue fibrosis with therapeutic potency to treat fibroproliferative diseases. © 2013 Lino Cardenas et al

Amyloid and tau pathology associations with personality traits, neuropsychiatric symptoms, and cognitive lifestyle in the preclinical phases of sporadic and autosomal dominant Alzheimer’s disease

Background Major prevention trials for Alzheimer’s disease (AD) are now focusing on multidomain lifestyle interventions. However, the exact combination of behavioral factors related to AD pathology remains unclear. In 2 cohorts of cognitively unimpaired individuals at risk of AD, we examined which combinations of personality traits, neuropsychiatric symptoms, and cognitive lifestyle (years of education or lifetime cognitive activity) related to the pathological hallmarks of AD, amyloid-β, and tau deposits. Methods A total of 115 older adults with a parental or multiple-sibling family history of sporadic AD (PREVENT-AD [PRe-symptomatic EValuation of Experimental or Novel Treatments for AD] cohort) underwent amyloid and tau positron emission tomography and answered several questionnaires related to behavioral attributes. Separately, we studied 117 mutation carriers from the DIAN (Dominant Inherited Alzheimer Network) study group cohort with amyloid positron emission tomography and behavioral data. Using partial least squares analysis, we identified latent variables relating amyloid or tau pathology with combinations of personality traits, neuropsychiatric symptoms, and cognitive lifestyle. Results In PREVENT-AD, lower neuroticism, neuropsychiatric burden, and higher education were associated with less amyloid deposition (p = .014). Lower neuroticism and neuropsychiatric features, along with higher measures of openness and extraversion, were related to less tau deposition (p = .006). In DIAN, lower neuropsychiatric burden and higher education were also associated with less amyloid (p = .005). The combination of these factors accounted for up to 14% of AD pathology. Conclusions In the preclinical phase of both sporadic and autosomal dominant AD, multiple behavioral features were associated with AD pathology. These results may suggest potential pathways by which multidomain interventions might help delay AD onset or progression

Biological responses of caged juvenile sea bass (Dicentrarchus labrax) and turbot (Scophtalmus maximus) in a harbour polluted area

International audienceField caging experiments were conducted on 0-group juvenileseabass (Dicentrarchuslabrax) and turbot (Scophtalmusmaximus) in apollutedarea. The fish were caged for 38 days in a northern French harbour (Boulogne sur Mer) at three locations with varying levels of contamination. Sediment from each location was analysed for PAHs, PCBs and metal contents, and their respective bioavailability. In the most heavily contaminated station, all the fish died during the two first weeks. At the two other stations, fish recovery was at least 75%. Growth and several health indices were individually measured on the tagged fish and metal bioaccumulation was measured in the fish gills. Juvenileseabass physiological performance showed significantly higher growth rates, RNA:DNA ratio and Fulton's K condition index in the least contaminated station. The growth in weight and the condition index of the turbotcaged in the less contaminated station were also higher. Similarly, for both species, lipid storage index based on the ratio of the quantity of triacylglycerols to sterols (TAG:ST), was significantly higher in the less contaminated station. This general decrease of growth and condition indices, associated with a metal accumulation in gills, reflected the adverse effects of contaminated sediment on the health of the fish. Species-specific responses to chemical contamination were recorded since a stronger relationship between physiological biomarkers and chemical contamination was observed in the seabass. Therefore, this study supports the use of seabass as model species and recommends measurements of physiological biomarkers for monitoring complex exposur

Identification of different Cu

Different copper-titanium oxides were obtained by impregnation of copper nitrate (atomic ratio Cu/Ti = 0.1) on three TiO2 samples with different initial structures: amorphous (AM), anatase (AN) and rutile (RT). X-band Electron Paramagnetic Resonance (EPR) analysis of Cu-TiO2 samples revealed the formation of different copper species depending on the initial structural state of TiO2. Aggregates of Cu(II) ions and isolated Cu2+ ions located either on the surface or in the solids were observed. Copper ions incorporated in anatase exhibit an octahedral symmetry mainly characterized by a significant elongation of Cu-O bonds following the z-axis. With the transformation of TiO2 from the anatase to the rutile form, copper atoms occupied Ti4+ sites in the rutile structure

Metal concentrations, growth and condition indices in European juvenile flounder (Platichthys flesus) relative to sediment contamination levels in four Eastern English Channel estuaries

International audienceThe aim of the present study was to evaluate the effects of metal contamination on the biological responses of 0-group juvenile European flounder and to assess and compare the quality of four estuarine habitats located in the Eastern English Channel. Fish otolith growth and condition indices (RNA : DNA ratio, Fulton's K condition index) were measured and found to be significantly lower in individuals from the Seine estuary compared to those of the Canche, Authie and Somme estuaries. No obvious effects of hydrological condition or food availability on the flounder biological responses were observed. Sediments from the Seine showed the highest metal concentrations, bioavailable proportion and enrichment factors. Higher metal concentrations were observed in fish from the Seine compared to the other ones caught in less polluted estuaries. These results suggest that contaminants may have a negative impact on the early life history stage of flounder

Synthesis and inclusion ability of a bis-beta-cyclodextrin pseudo-cryptand towards Busulfan anticancer agent

The synthesis of a C2-symmetric receptor including two β-cyclodextrins connected by urea linkers to a chiral diaza-crown ether organising platform is reported. This molecular system, long thought to be a potent selective carrier for chiral/achiral organic/inorganic guests at the supramolecular level, was found to be an efficient complexing tool towards the Busulfan anticancer agent

Growth and condition indices of juvenile turbot, Scophthalmus maximus, exposed to contaminated sediments: Effects of metallic and organic compounds.

International audienceSince sediments have the potential to form associations with several classes of pollutants, they have been recognized as a possible and significant source of contamination for the benthic environment. Flatfish maintain a close association with sediments for food and cover, and are therefore more likely to be exposed to contaminated sediments, especially in coastal areas (e.g. nursery grounds). The assessment of these potential biological effects involves the use of adapted biomonitoring tools. The main objective of this study was to assess and compare the response of several physiological biomarkers measured on juvenile turbot (Scophthalmus maximus) exposed to contaminated sediments. Sediments were collected from three stations in a harbour in northern France (Boulogne-sur-Mer), in an anthropogenic French estuary (the Seine), and in a reference site (exposed sandy beach of Wimereux). Unexposed lab-reared juvenile turbots were exposed to sediments for 7 and 21 days in laboratory conditions. Sediments were analysed for metals, PAH and PCB contamination. Several fish growth and condition indices were individually analysed in fish according to the chemical contaminant availability in sediment, the metal concentrations in gills and the estimation of PAH metabolites in their bile. Significant decreases in growth rates, morphometric index, RNA:DNA ratio and the lipid storage index, based on the ratio of the quantity of triacylglycerols on sterols (TAG:ST), were observed with increasing level of chemical contamination. This decrease in the fish's physiological status could be related to the significant increase of several metal concentrations in contaminated fish gills and the significant increase of PAH metabolites in bile. In a field situation, such a reduction in growth and energetic status of juvenile fish could dramatically decrease their over-winter survival in contaminated nursery grounds