ABSTRACT
Enhancing brown fat activity and promoting white fat browning are attractive therapeutic strategies for treating obesity and associated metabolic disorders. To provide a comprehensive picture of the gene regulatory network in these processes, we conducted a series of transcriptome studies by RNA sequencing (RNA-seq) and quantified the mRNA and long noncoding RNA (lncRNA) changes during white fat browning (chronic cold exposure, beta-adrenergic agonist treatment, and intense exercise) and brown fat activation or inactivation (acute cold exposure or thermoneutrality, respectively). mRNA-lncRNA coexpression networks revealed dynamically regulated lncRNAs to be largely embedded in nutrient and energy metabolism pathways. We identified a brown adipose tissue-enriched lncRNA, lncBATE10, that was governed by the cAMP-cAMP response element-binding protein (Creb) axis and required for a full brown fat differentiation and white fat browning program. Mechanistically, lncBATE10 can decoy Celf1 from Pgc1α, thereby protecting Pgc1α mRNA from repression by Celf1. Together, these studies provide a comprehensive data framework to interrogate the transcriptomic changes accompanying energy homeostasis transition in adipose tissue.
Subject(s)
Adipose Tissue/metabolism , CELF1 Protein/metabolism , Energy Metabolism , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha/metabolism , RNA, Long Noncoding/metabolism , Animals , Cell Differentiation , Cyclic AMP/metabolism , Cyclic AMP Response Element-Binding Protein/metabolism , Gene Expression Regulation , HEK293 Cells , Humans , Male , Mice, Inbred C57BL , Primary Cell Culture , Signal Transduction , TranscriptomeABSTRACT
Human antigen R (HuR) is an essential regulator of RNA metabolism, but its function in metabolism remains unclear. This study identifies HuR as a major repressor during adipogenesis. Knockdown and overexpression of HuR in primary adipocyte culture enhances and inhibits adipogenesis in vitro, respectively. Fat-specific knockout of HuR significantly enhances adipogenic gene program in adipose tissues, accompanied by a systemic glucose intolerance and insulin resistance. HuR knockout also results in depot-specific phenotypes: it can repress myogenesis program in brown fat, enhance inflammation program in epidydimal white fat and induce browning program in inguinal white fat. Mechanistically, HuR may inhibit adipogenesis by recognizing and modulating the stability of hundreds of adipocyte transcripts including Insig1, a negative regulator during adipogenesis. Taken together, our work establishes HuR as an important posttranscriptional regulator of adipogenesis and provides insights into how RNA processing contributes to adipocyte development.
Subject(s)
Adipogenesis/genetics , Adipogenesis/physiology , ELAV-Like Protein 1/metabolism , RNA-Binding Proteins/genetics , RNA-Binding Proteins/metabolism , Adipose Tissue/pathology , Adipose Tissue, Brown/metabolism , Adipose Tissue, White/metabolism , Animals , ELAV-Like Protein 1/genetics , Gene Expression Regulation , Gene Knockout Techniques , Glucose Intolerance/metabolism , Humans , Inflammation , Insulin Resistance , Intracellular Signaling Peptides and Proteins , Male , Membrane Proteins , Mice, Inbred C57BL , Mice, KnockoutABSTRACT
Obesity induces profound transcriptome changes in adipocytes, and recent evidence suggests that long-noncoding RNAs (lncRNAs) play key roles in this process. We performed a comprehensive transcriptome study by RNA sequencing in adipocytes isolated from interscapular brown, inguinal, and epididymal white adipose tissue in diet-induced obese mice. The analysis revealed a set of obesity-dysregulated lncRNAs, many of which exhibit dynamic changes in the fed versus fasted state, potentially serving as novel molecular markers of adipose energy status. Among the most prominent lncRNAs is Lnc-leptin, which is transcribed from an enhancer region upstream of leptin (Lep). Expression of Lnc-leptin is sensitive to insulin and closely correlates to Lep expression across diverse pathophysiological conditions. Functionally, induction of Lnc-leptin is essential for adipogenesis, and its presence is required for the maintenance of Lep expression in vitro and in vivo. Direct interaction was detected between DNA loci of Lnc-leptin and Lep in mature adipocytes, which diminished upon Lnc-leptin knockdown. Our study establishes Lnc-leptin as a new regulator of Lep.
Subject(s)
Adipocytes, Brown/metabolism , Adipocytes, White/metabolism , Adipogenesis , Gene Expression Regulation , Leptin/agonists , Obesity/metabolism , RNA, Long Noncoding/metabolism , Adipocytes, Brown/drug effects , Adipocytes, Brown/pathology , Adipocytes, White/drug effects , Adipocytes, White/pathology , Adipogenesis/drug effects , Animals , Base Sequence , Biomarkers/metabolism , Cells, Cultured , Diet, High-Fat/adverse effects , Energy Metabolism/drug effects , Enhancer Elements, Genetic/drug effects , Gene Expression Profiling , Gene Expression Regulation/drug effects , Gene Ontology , Hypoglycemic Agents/pharmacology , Insulin/pharmacology , Leptin/antagonists & inhibitors , Leptin/genetics , Leptin/metabolism , Male , Mice, Inbred C57BL , Obesity/etiology , Obesity/pathology , RNA Interference , RNA, Long Noncoding/antagonists & inhibitors , RNA, Long Noncoding/chemistry , RNA, Small Interfering/metabolismABSTRACT
Obesity has emerged as an alarming health crisis due to its association with metabolic risk factors such as diabetes, dyslipidemia, and hypertension. Recent work has demonstrated the multifaceted roles of lncRNAs in regulating mouse adipose development, but their implication in human adipocytes remains largely unknown. Here we present a catalog of 3149 adipose active lncRNAs, of which 909 are specifically detected in brown adipose tissue (BAT) by performing deep RNA-seq on adult subcutaneous, omental white adipose tissue and fetal BATs. A total of 169 conserved human lncRNAs show positive correlation with their nearby mRNAs, and knockdown assay supports a role of lncRNAs in regulating their nearby mRNAs. The knockdown of one of those, lnc-dPrdm16, impairs brown adipocyte differentiation in vitro and a significant reduction of BAT-selective markers in in vivo. Together, our work provides a comprehensive human adipose catalog built from diverse fat depots and establishes a roadmap to facilitate the discovery of functional lncRNAs in adipocyte development.