Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of mouse liver exposed to 24h cytokine starvation vs. treatment with TNF alpha

ABSTRACT: Pathogenesis of atherosclerosis results from the interactions between disrupted lipid homeostasis and immune response but the molecular bridges between the major players are still a matter of controversy. We investigated the systemic effect of inflammatory cytokine tumor necrosis factor alpha (TNF-Î±), a well established anorexia agent, in livers; of mice exposed to 24h-cytokine/starvation. 10% of genes of the Agilent 10kb cDNA array is detected above level of detection in the mouse liver. 12% of these is modulated upon starvation while 22% is altered by TNF-Î±. Experiment Overall Design: Twelve mice (C57BL/6, Harlan) were randomly assigned into three groups. Three animals were i.v. injected by human recombinant TNF-Î± (30 Âµg/animal), a dose that is known to have a significant effect in Experiment Overall Design: mice without leading to mortality. Food was withdrawn after application of TNF-alpha (TNF/starvation group). Further, for three animals which were treated with corresponding volume (200Âµl) of saline a food was withdrawn (starvation group). Three animals from control group had free access to food throughout the study. Mice were sacrificed 20 hours later. Livers for RNA analysis were stored in RNAlater (Quiagen) according to manufacture's instruction. RNA of individual treated mouse was hybridized versus control sample which is obtained by pooling of control RNAs.

ORGANISM(S): Mus musculus

SUBMITTER: Drago Kuzman

PROVIDER: E-GEOD-6317 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

The Sterolgene v0 cDNA microarray: a systemic approach to studies of cholesterol homeostasis and drug metabolism.

Rezen Tadeja T Juvan Peter P Fon Tacer Klementina K Kuzman Drago D Roth Adrian A Pompon Denis D Aggerbeck Lawrence P LP Meyer Urs A UA Rozman Damjana D

BMC genomics 20080211

<h4>Background</h4>Cholesterol homeostasis and xenobiotic metabolism are complex biological processes, which are difficult to study with traditional methods. Deciphering complex regulation and response of these two processes to different factors is crucial also for understanding of disease development. Systems biology tools as are microarrays can importantly contribute to this knowledge and can also discover novel interactions between the two processes.<h4>Results</h4>We have developed a low den ...[more]

PMID: 18261244

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Project description:Schistosoma mansoni is the major causative agent of schistosomiasis in the Americas. This parasite takes advantage from host signaling molecules such as cytokines and hormones to complete its development inside the host. TNF-Î± is the most important cytokine involved in the inflammatory response when cercaria, the infective stage, penetrates the human skin and a severe inflammatory response is started. In this work the authors describe the complete sequence of a possible TNF-Î± receptor in S. mansoni and detect that the receptor is most highly expressed in cercaria among all life cycle stages. In an attempt to mimic the situation at the site of skin penetration cercariae have been mechanically transformed in vitro into schistosomula and immediately exposed to human TNF-Î± . Exposure of these early schistosomula to the human hormone caused a large-scale change in the expression of parasite genes. Exposure of adult worms to human TNF-Î± caused gene expression changes as well, although the set of parasite altered genes was entirely different from that of schistosomula. This work increases the number of known signaling pathways of the parasite, and opens new perspectives into understanding the molecular components of TNF-Î± response as well as possibly interfering with parasite-host interaction. A platform previously designed by our group (PMID: 17517391) was used. 300 ng of RNA from adult schistosomes treated with TNF and its control were used for the hybridization. RNA amplification and labeling kit (Agilent Technologies) were used according to manufacturerÂ´s instructions. Each sample was separately labeled with either Cy3 or Cy5. 825 ng cRNA from each amplification were used for hybridization; self-self experiments were performed. Washing and scanning procedures were according to the manufacturerÂ´s instructions using GenePix 4000B scanner (Molecular Devices, Sunnyvale, CA, USA). Data was extracted using Feature Extraction software (Agilent Technologies). We calculated a virtual Log2 ratio between intensities of TNF-Î± Treated/Control, for each gene. With these ratios, we used two different approaches for SAM statistical analyses. SAM one-class approach was used to identify genes with sustained changes in their expression levels along the entire time period of observation (1 and 24 h); SAM two-class approach was used to identify genes with transient changes in their expression levels. In both cases, genes were considered differentially expressed at q-value â?¤ 0.05. Hierarchical clustering of selected genes was generated using Spotfire Decision Site software (TIBCO Software Inc., Palo Alto, CA, USA). For a gene that was represented in the array by multiple probes, we picked a single representative probe by selecting the probe with the highest absolute value of the Log2 ratio between intensities of TNF-Î± Treated/Control.

Project description:Acute respiratory distress syndrome (ARDS) is a catastrophic form of acute lung injury (ALI). The necessity for mechanical ventilation (MV) renders patients at risk for ventilator induced lung injury (VILI). Exposure to repetitive cyclic stretch (CS) and/or over-inflation exacerbates injury. Reducing tidal volume (VT) is the only therapeutic strategy shown to mitigate morbidity and mortality. Cyclic stretch has been shown to differentially regulate gene expression in part through the activation of mammalian mitogen-activated protein kinase (MAPK). Although these studies have shown both molecular and cellular alterations, no unifying hypothesis to explain MV-induced lung injury has emerged. In the current study, we hypothesized that coordinated expression of cyclic stretch (CS)-responsive genes relies on the presence of common CS-sensitive regulatory elements. To identify CS-responsive genes, we undertook a comparative examination of the gene expression profile of human bronchial epithelial airway (Beas-2B) cells in response to various injurious stimuli involved in the pathogenesis of acute lung injury (ALI)/Ventilator induced lung injury (VILI): cyclic stretch, tumor necrosis factor alpha (TNF-a), and lipopolysaccharide (LPS). Experiment Overall Design: Human Bronchial Epithelial Cells (Beas-B2) cells grown on silicon elastic plates coated with Type I collagen (Flexercell International, McKeesport, PA) were exposed to six regiments for 4 h: 1) control (static, [control]); 2) mechanical stretch (25 PKa, 30 cycles per min, [stretch]); 3) LPS (1 mcg/ml [LPS]); 4) TNF-Î± (20 ng/ml; [TNF]); 5) mechanical stretch plus LPS [LPS+S], and 6) mechanical stretch plus TNF-Î± [TNF+S]. Total RNA (duplicate experiments) was extracted using TRIZOL reagent (as per manufactures specifications) and purified using Qiagen mRNA purification Kit (as per manufacturers specifications). mRNA was hybridized to Affymetrix Human U133plus2.0 chips. Probe based analysis, background reduction, and quantile data normalization was performed in MeV 4.0 of TM4 using Robust Multi-array Average (RMA).

Dataset Information

Transcription profiling of mouse liver exposed to 24h cytokine starvation vs. treatment with TNF alpha

Publications

The Sterolgene v0 cDNA microarray: a systemic approach to studies of cholesterol homeostasis and drug metabolism.

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