Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcriptomic profile of human preadipocyte treated by macrophages from adipose tissue

ABSTRACT: Obesity is characterized by a chronic auto-inflammation of the hypertrophied white adipose tissue (WAT) associated with metabolic and vascular complications. WAT of obese subjects produces inflammatory factors like cytokines (IL6, TNF-alpha) and chemokines (CCL2 and IL-8) originating mainly from the accumulated macrophages. WAT macrophages profoundly affect adipose cell biology leading in particular to the inflammation of the adipocytes and altered differentiation of their precursors, the pre-adipocytes. We previously showed that conditioned media from WAT macrophages (ATM CM) induced inflammation in human pre-adipocytes. We performed pangenomic cDNA microarrays analysis to compare gene expression profiles between control and ATM CM treated preadipocytes. By a functional analysis, we identified the biological themes that best characterized the proinflammatory preadipocytes. The most significantly overrepresented GO Cellular Component categories were "cytokine-cytokine receptor interaction" and "chemokine signaling pathway", emphasizing the importance of cytokine and chemokine production by the inflammatory preadipocyte. The chemokines acting in concert to recruit leukocytes in inflamed tissues, still need to be more precisely identified in human WAT. Thus, our data lead to the identification of new chemokines involved in attraction of immune cells in inflamed human WAT. The transcriptomic profile of human preadipocytes was compared to that of preadipocyes treated by macrophages isolated from human adipose tissue. Two-condition experiment,human control preadipocyte vs preadipocyte treated by macrophages from adipose tissue. 5 biological replicates (with a dye-swap design).

ORGANISM(S): Homo sapiens

SUBMITTER: DaniM-CM-(le Lacasa

PROVIDER: E-GEOD-28445 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Roles of chemokine ligand-2 (CXCL2) and neutrophils in influencing endothelial cell function and inflammation of human adipose tissue.

Rouault Christine C Pellegrinelli Vanessa V Schilch Raphaela R Cotillard Aurélie A Poitou Christine C Tordjman Joan J Sell Henrike H Clément Karine K Lacasa Danièle D

Endocrinology 20130131 3

The hypertrophied white adipose tissue (WAT) during human obesity produces inflammatory mediators, including cytokines (IL-6 and TNFα) and chemokines ([C-C motif] chemokine ligand 2 and IL-8). These inflammatory factors are preferentially produced by the nonadipose cells, particularly the adipose tissue infiltrating macrophages. We identified the chemokine (C-X-C motif) ligand 2 (CXCL2) by a transcriptomic approach. Because CXCL2 could represent a WAT-produced chemokine, we explored its role in ...[more]

PMID: 23372021

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Project description:Full title: Expression data from human primary subcutaneous preadipocytes treated with glucocorticoids prior to the initiation of differentiation. Preadipocytes are continuously exposed to glucocorticoids in situ due to both steroid present in the circulatory system as well as adipose tissue specific 11Î²HSD1 activity. While the effects of glucocorticoids during differentiation are well studied, the effect of exposure of preadipocytes to glucocorticoids prior to differentiation is unknown. We therefore treated confluent human primary preadipocytes drived from subcutaneous adipose tissue with the synthetic glucocorticoid dexamethasone for 48 hours prior to the initiation of differentiation and assessed what effect this had on their subsequent potential to differentiate. We found that pretreatment with glucocorticoids had a priming effect and resulted in increased differentiation of these preadipocytes. Furthermore, this treatment was additive to the effects of glucocorticoids during the initial phase of adipogenesis. Microarray analysis performed subsequent to the pretreatment with glucocorticoids (at the time point at which preadipocytes would have been induced to differentiate) identified glucocorticoid-responsive, candidate genes whose altered expression could mediate these effects. keywords: glucocorticoids, glucocorticoid receptor, preadipocytes, adipogenesis, human primary preadipocytes, subcutaneous, adipose tissue Experiment Overall Design: Human subcutaneous primary preadipocytes were purchased from Zen-Bio Inc. Preadipocytes from 5 female donors (average BMI 22.5Â±0.2kg/m2) were pooled prior to the initial seeding in T75 flasks. Cells were maintained at 5% CO2 in DMEM with 1.0g/L glucose, 20% calf serum, 100U/ml penicillin, 100mg/ml streptomycin and 50U/ml nystatin. Cells were expanded once prior to seeding in Nunc-brand 12-well dishes. Upon reaching confluence (24 h post-splitting), preadipocytes were stimulated with vehicle or 1uM dex for 48 hours in growth media containing 3% calf serum. Microarray analysis was performed on duplicate samples.

Project description:Differentiation of 3T3-L1 preadipocytes can be induced by a 2-d treatment with a factor "cocktail" (DIM) containing the synthetic glucocorticoid dexamethasone (dex), insulin, the phosphodiesterase inhibitor methylisobutylxanthine (IBMX) and fetal bovine serum (FBS). We temporally uncoupled the activities of the four DIM components and found that treatment with dex for 48 h followed by IBMX treatment for 48 h was sufficient for adipogenesis, whereas treatment with IBMX followed by dex failed to induce significant differentiation. Similar results were obtained with C3H10T1/2 and primary mesenchymal stem cells. The 3T3-L1 adipocytes differentiated by sequential treatment with dex and IBMX displayed insulin sensitivity equivalent to DIM adipocytes, but had lower sensitivity to isoproterenol-stimulated lipolysis and reduced triglyceride content. The nondifferentiating IBMX–then-dex treatment produced transient expression of adipogenic transcriptional regulatory factors C/EBP{beta} and C/EBP{delta}, and little induction of terminal differentiation factors C/EBP{alpha} and PPAR{gamma}. Moreover, the adipogenesis inhibitor preadipocyte factor-1 (Pref-1) was repressed by DIM or by dex-then-IBMX, but not by IBMX-then-dex treatment. We conclude that glucocorticoids drive preadipocytes to a novel intermediate cellular state, the dex-primed preadipocyte, during adipogenesis in cell culture, and that Pref-1 repression may be a cell fate determinant in preadipocytes. We sought to define the genes differentially expressed in response to dexamethasone (dex) in 3T3L1 mouse preadipocytes. We treated 3T3L1 cells with dex or DMSO vehicle for 2 or 36 hrs and then prepared total RNA. We hybridized each sample (Cy5) to a pool of the samples, which served as the reference (Cy3) channel. All linear models for differential gene expression were constructed with the "limma" package (Smyth, 2004) in BioConductor (Gentleman et al., 2004). A linear model for dex was constructed to compare the 4 biological replicates of DMSO vehicle treatment with dex treatment at either 2 or 36hr. Genes were considered differentially expressed if at least one probe had log-odds greater than 50:50 in the linear model.

Dataset Information

Transcriptomic profile of human preadipocyte treated by macrophages from adipose tissue

Publications

Roles of chemokine ligand-2 (CXCL2) and neutrophils in influencing endothelial cell function and inflammation of human adipose tissue.

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