Dataset Information

Transcriptome Modifications in Porcine Adipocytes via Toll-Like Receptors Activation.

ABSTRACT: Adipocytes are the most important cell type in adipose tissue playing key roles in immunometabolism. We previously reported that nine members of the Toll-like receptor (TLR) family are expressed in an originally established porcine intramuscular pre-adipocyte (PPI) cell line. However, the ability of TLR ligands to modulate immunometabolic transcriptome modifications in porcine adipocytes has not been elucidated. Herein, we characterized the global transcriptome modifications in porcine intramuscular mature adipocytes (pMA), differentiated from PPI, following stimulation with Pam3csk4, Poly(I:C) or LPS which are ligands for TLR2, TLR3, and TLR4, respectively. Analysis of microarray data identified 530 (218 up, 312 down), 520 (245 up, 275 down), and 525 (239 up, 286 down) differentially expressed genes (DEGs) in pMA following the stimulation with Pam3csk4, Poly(I:C), and LPS, respectively. Gene ontology classification revealed that DEGs are involved in several biological processes including those belonging to immune response and lipid metabolism pathways. Functionally annotated genes were organized into two groups for downstream analysis: immune response related genes (cytokines, chemokines, complement factors, adhesion molecules, and signal transduction), and genes involved with metabolic and endocrine functions (hormones and receptors, growth factors, and lipid biosynthesis). Differential expression analysis revealed that EGR1, NOTCH1, NOS2, TNFAIP3, TRAF3IP1, INSR, CXCR4, PPARA, MAPK10, and C3 are the top 10 commonly altered genes of TLRs induced transcriptional modification of pMA. However, the protein-protein interaction network of DEGs identified EPOR, C3, STAR, CCL2, and SAA2 as the major hub genes, which were also exhibited higher centrality estimates in the Gene-Transcription factor interaction network. Our results provide new insights of transcriptome modifications associated with TLRs activation in porcine adipocytes and identified key regulatory genes that could be used as biomarkers for the evaluation of treatments having immunomodularoty and/or metabolic functional beneficial effects in porcine adipocytes.

SUBMITTER: Igata M

PROVIDER: S-EPMC6549529 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Transcriptome Modifications in Porcine Adipocytes via Toll-Like Receptors Activation.

Igata Manami M Islam Md Aminul MA Tada Asuka A Takagi Michihiro M Kober A K M Humayun AKMH Albarracin Leonardo L Aso Hisashi H Ikeda-Ohtsubo Wakako W Miyazawa Kenji K Yoda Kazutoyo K He Fang F Takahashi Hideki H Villena Julio J Kitazawa Haruki H

Frontiers in immunology 20190529

Adipocytes are the most important cell type in adipose tissue playing key roles in immunometabolism. We previously reported that nine members of the Toll-like receptor (TLR) family are expressed in an originally established porcine intramuscular pre-adipocyte (PPI) cell line. However, the ability of TLR ligands to modulate immunometabolic transcriptome modifications in porcine adipocytes has not been elucidated. Herein, we characterized the global transcriptome modifications in porcine intramusc ...[more]

PMID: 31191544

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Project description:Adipocytes are dynamic cells that have critical functions to maintain body energy homeostasis. Function of adipocyte is affected by adipogenic differentiation, and exogenous stimulation of biochemical factors such as serotonin and TNF-. Serotonin (5-hydroxytriptamine, 5-HT) is a biogenic monoamine, which known to be involved in lipid metabolism in adipocytes. The TNF- is a proinflammatory cytokines involved in adipocyte biology. Here we investigated the global transcriptome alterations of porcine intramuscular adipocytes at undifferentiated preadipocyte, differentiated adipocytes state, and adipocytes after stimulation with serotonin and TNF- for 12 h by using Agilent’s Porcine (V2) one-color Gene Expression Microarray 4 × 44. Transcriptome analysis revealed that the expression of 443, 261, and 249 genes were altered after differentiation, and after serotonin and TNF- stimulation, respectively. Differentiation process of adipocytes are found to be involved with expression changes of APP, HNF4A, ESR1, EGR1, SRC, HNF1A, FN1, ALB, STAT3, CBL, CEBPB, AR, FOS, CFTR, PAN2, PTPN6, VDR, PPARG, STAT5A and NCOA3 genes which are enriched in the ‘PPAR signaling pathway’ and ‘insulin resistance pathway’. Dose-dependent serotonin stimulation resulted in a decreased fat accumulation in the adipocytes as well as decreased adipocyte proliferation. Serotonin-induced differentially expressed genes in adipocytes were participated in the significant enrichment of GPCR ligand-binding, Cell chemotaxis, blood coagulation and complement pathways, metabolism of lipid and lipoproteins, regulation of lipid metabolism by PPARA; and lipid digestion, mobilization and transport pathways. TNF-α-induced transcriptional modification follows the similar trend, and alteration of many genes shared between both serotonin- and TNF-α stimulations. Our results provide new insights of transcriptome modifications associated with adipogenesis, and exogenous stimulation of serotonin- and TNF- to the porcine intramuscular adipocytes.

Project description:BackgroundCritically ill patients frequently develop muscle atrophy and weakness in the intensive-care-unit setting [intensive care unit-acquired weakness (ICUAW)]. Sepsis, systemic inflammation, and acute-phase response are major risk factors. We reported earlier that the acute-phase protein serum amyloid A1 (SAA1) is increased and accumulates in muscle of ICUAW patients, but its relevance was unknown. Our objectives were to identify SAA1 receptors and their downstream signalling pathways in myocytes and skeletal muscle and to investigate the role of SAA1 in inflammation-induced muscle atrophy.MethodsWe performed cell-based in vitro and animal in vivo experiments. The atrophic effect of SAA1 on differentiated C2C12 myotubes was investigated by analysing gene expression, protein content, and the atrophy phenotype. We used the cecal ligation and puncture model to induce polymicrobial sepsis in wild type mice, which were treated with the IкB kinase inhibitor Bristol-Myers Squibb (BMS)-345541 or vehicle. Morphological and molecular analyses were used to investigate the phenotype of inflammation-induced muscle atrophy and the effects of BMS-345541 treatment.ResultsThe SAA1 receptors Tlr2, Tlr4, Cd36, P2rx7, Vimp, and Scarb1 were all expressed in myocytes and skeletal muscle. Treatment of differentiated C2C12 myotubes with recombinant SAA1 caused myotube atrophy and increased interleukin 6 (Il6) gene expression. These effects were mediated by Toll-like receptors (TLR) 2 and 4. SAA1 increased the phosphorylation and activity of the transcription factor nuclear factor 'kappa-light-chain-enhancer' of activated B-cells (NF-κB) p65 via TLR2 and TLR4 leading to an increased binding of NF-κB to NF-κB response elements in the promoter region of its target genes resulting in an increased expression of NF-κB target genes. In polymicrobial sepsis, skeletal muscle mass, tissue morphology, gene expression, and protein content were associated with the atrophy response. Inhibition of NF-κB signalling by BMS-345541 increased survival (28.6% vs. 91.7%, P < 0.01). BMS-345541 diminished inflammation-induced atrophy as shown by a reduced weight loss of the gastrocnemius/plantaris (vehicle: -21.2% and BMS-345541: -10.4%; P < 0.05), tibialis anterior (vehicle: -22.7% and BMS-345541: -17.1%; P < 0.05) and soleus (vehicle: -21.1% and BMS-345541: -11.3%; P < 0.05) in septic mice. Analysis of the fiber type specific myocyte cross-sectional area showed that BMS-345541 reduced inflammation-induced atrophy of slow/type I and fast/type II myofibers compared with vehicle-treated septic mice. BMS-345541 reversed the inflammation-induced atrophy program as indicated by a reduced expression of the atrogenes Trim63/MuRF1, Fbxo32/Atrogin1, and Fbxo30/MuSA1.ConclusionsSAA1 activates the TLR2/TLR4//NF-κB p65 signalling pathway to cause myocyte atrophy. Systemic inhibition of the NF-κB pathway reduced muscle atrophy and increased survival of septic mice. The SAA1/TLR2/TLR4//NF-κB p65 atrophy pathway could have utility in combatting ICUAW.

Dataset Information

Transcriptome Modifications in Porcine Adipocytes via Toll-Like Receptors Activation.

Publications

Transcriptome Modifications in Porcine Adipocytes via Toll-Like Receptors Activation.

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