Dysregulation of a long noncoding RNA reduces leptin leading to a leptin-responsive form of obesity

Dallner, Olof S.; Marinis, Jill M.; Lu, Yi-Hsueh; Birsoy, Kıvanç; Werner, Emory; Fayzikhodjaeva, Gulya; Dill, Brian D.; Molina, Henrik; Moscati, Arden; Kutalik, Zoltán; Marques‐Vidal, Pedro; Kilpeläinen, Tuomas O.; Grarup, Niels; Linneberg, Allan; Zhang, Yinxin; Vaughan, Roger; Loos, Ruth J.F.; Lazar, Mitchell A.; Friedman, Jeffrey M.

doi:10.1038/s41591-019-0370-1

Cited by 92 publications

(84 citation statements)

References 43 publications

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“…Previous studies from our laboratory have shown that redundant sequences in the extreme 5' and 3' regions of the gene, greater than 10kb from the TSS, can confer fatspecific expression of the leptin gene (18,19). However, because mutations in these regions interfered with the fat-specific expression of this gene, it was impossible to define the sequences that quantitatively regulated leptin expression.…”

Section: Discussionmentioning

confidence: 98%

“…, is Homologous to LepRE1. LE1, located between -16.5 to -16.1kb, and LE2, located between +13.6 to +13.9 kb, are redundant elements that can independently confer qualitative and quantitative expression of luciferase in leptin-reporter mice --raising the possibility that functionally similar elements at both sites can regulate the level of leptin gene expression (18,19). In previous studies, deletion of LE2 in a BAGTG extending between -0.762 to +18 kb ablated fat specific expression of a luciferase reporter.…”

Section: A 3' Fat-regulated Footprint Lepre2mentioning

confidence: 99%

“…We also found that a 3' BACTG extending from -762bp to +18kb was able to drive reporter expression in a manner similar to the 5' 31kb (-22 to +8.8 kb) BACTG, while a BACTG extending from -762bp to 8.8kb lost fat-specific reporter expression. Subsequent studies have identified two redundant elements that can independently confer cell-specific expression of leptin in adipocytes; LE1 is localized between -16.5 and -16.1 kb upstream of the leptin TSS while LE2 is located between + 13.6 and +13.9 kb downstream of the leptin TSS (19). However, because deletion of these elements ablates fat-specific expression altogether, it is not possible to assess the role of these specific sequences to quantitatively regulate leptin expression.…”

Section: Introductionmentioning

confidence: 99%

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A non-canonical-PPARγ/RXRα-binding sequence regulates leptin expression in response to changes in adipose tissue mass

Zhang

Dallner

Nakadai

et al. 2018

Preprint

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Leptin expression decreases after fat loss and is increased when obesity develops and its proper quantitative regulation is essential for the homeostatic control of fat mass. We previously reported that a distant leptin enhancer (LE1), 16kb upstream from the transcription start site (TSS), confers fat-specific expression in a BAC transgenic reporter mouse (BACTG). However this and the other elements that we identified do not account for the quantitative changes in leptin expression that accompany alterations of adipose mass. In this report, we used ATAC-seq to identify a 17bp non-canonical-PPARγ/RXRα-binding site leptin regulatory element 1 (LepRE1) within LE1, and show that it is necessary for the fat-regulated quantitative control of reporter (luciferase) expression. While BACTG reporter mice with mutations in this sequence still show fatspecific expression, luciferase is no longer decreased after food restriction and weight loss. Similarly the increased expression of leptin reporter associated with obesity in ob/ob mice is impaired. A functionally analogous LepRE1 site is also found in a second, redundant DNA regulatory element 13kb downstream of the TSS. These data uncouple the mechanisms conferring qualitative and quantitative expression of the leptin gene and further suggest that factor(s) that bind to LepRE1 quantitatively control leptin expression and might be components of a lipid sensing system in adipocytes.

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Section: Discussionmentioning

confidence: 98%

Section: A 3' Fat-regulated Footprint Lepre2mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A non-canonical-PPARγ/RXRα-binding sequence regulates leptin expression in response to changes in adipose tissue mass

Zhang

Dallner

Nakadai

et al. 2018

Preprint

Self Cite

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“…Among the top-ranked miR-29 targets, we focused on leptin due to its central role in regulating energy homeostasis. Recent work has identified control of leptin transcription through an enhancer region 16 kb upstream of the transcriptional start site 31,32 , but until now, no post-transcriptional regulators of leptin have been described. This is surprising since miRNAs are estimated to regulate a majority of mRNAs in mammals 33 , and the leptin 3’-UTR is roughly 3-fold the length of an average 3’-UTR in vertebrates 34 .…”

Section: Figmentioning

confidence: 99%

AGO HITS-CLIP in Adipose Tissue Reveals miR-29 as a Post-Transcriptional Regulator of Leptin

O’Connor

Murphy

et al. 2020

Preprint

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MicroRNAs (miRNAs) are short, non-coding RNAs that associate with Argonaute (AGO) to regulate mRNA stability and translation. While individual miRNAs have been shown to play important roles in white and brown adipose tissue in normal physiology and disease 1,2,3 , a comprehensive analysis of miRNA activity in these tissues has not been performed. We used high-throughput sequencing of RNA isolated by crosslinking immunoprecipitation (HITS-CLIP) to comprehensively characterize the network of high-confidence, in vivo mRNA:miRNA interactions across white and brown fat, revealing over 100,000 unique miRNA binding sites. Targets for each miRNA were ranked to generate a catalog of miRNA binding activity, and the miR-29 family emerged as a top regulator of adipose tissue gene expression. Among the top targets of miR-29 was leptin, an adipocyte-derived hormone that acts on the brain to regulate food intake and energy expenditure 4 . Two independent miR-29 binding sites in the leptin 3'-UTR were validated using luciferase assays, and miR-29 gain and loss-of-function modulated leptin mRNA and protein secretion in primary adipocytes. In mice, miR-29 abundance inversely correlated with leptin levels in two independent models of obesity. This work represents the only experimentally generated miRNA targetome in adipose tissue and identifies the first known post-transcriptional regulator of leptin. Future work aimed at manipulating miR-29:leptin binding may provide a therapeutic opportunity to treat obesity and its sequelae.Much of the work on adipocyte development and function over the last several decades has focused on transcriptional regulators. These include PPARG2, which is necessary and sufficient for the development and maintenance of white and brown adipocytes 5 , and PRDM16, which regulates brown and beige adipocyte identity 6,7 . More recently, it has become clear that post-transcriptional regulators also play 3 a key role in influencing adipocyte phenotype. Studies with adipose-specific dicer KO mice demonstrate that microRNAs (miRNAs) are essential in regulating adipogenesis, insulin sensitivity and non-shivering thermogenesis 8,9 . Several individual miRNAs have been shown to contribute to these phenotypic effects.miR-133 represses BAT function by directly targeting Prdm16 and inhibiting expression of thermogenic genes 2 . miR-196a induces browning of WAT by targeting Hoxc8 1 , and miR-26 protects mice from dietinduced obesity by blocking adipogenesis 10 . These studies, and most others in adipose tissue, rely on computational predictions and low-throughput validation to identify miRNA targets. Although highthroughput crosslinking techniques to map the miRNA targetome have been applied to liver, brain, heart and other tissues 11,12,13 , thus far, no comprehensive targetome has been described for adipose tissue.To profile the complete network of in vivo, adipose-specific miRNA binding activity across both brown and white fat, we used AGO HITS-CLIP on interscapular brown adipose tissue (iBAT) and epididymal white a...

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“…For example, lncRNA Blnc1 works in concert with another important hepatic nuclear receptor, liver X receptor (LXR), to activate the lipogenic gene program (Zhao et al, 2018). Down-regulation of lncRNA lncOb reduces leptin, leading to a leptin responsive form of obesity (Dallner et al, 2019). Other studies found that lncRNAs are regulated during adipocyte differentiation (Yuan et al, 2019a;Yuan et al, 2019b), are expressed in liver in a sex-dependent manner when regulated by growth hormone (Melia et al, 2016;Melia and Waxman, 2019), and can be strongly induced by xenobiotic exposure (Lodato et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Long non-coding RNA Gm15441 attenuates hepatic inflammasome activation in response to metabolic stress

Brocker

Kim

Melia

et al. 2019

Preprint

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Fasting paradigms elicit a wide-range of health benefits including suppressing inflammation.Exploring the molecular mechanisms that prevent inflammation during caloric restriction may yield promising new therapeutic targets. During fasting, activation of the nuclear receptor peroxisome proliferator-activated receptor alpha (PPARA) promotes the utilization of lipids as an energy source. Herein, we show that ligand activation of PPARA directly upregulates the long non-coding RNA gene Gm15441 through binding sites within its promoter. Gm15441 expression suppresses its antisense transcript, encoding thioredoxin interacting protein (TXNIP). This, in turn, decreases TXNIP-stimulated NLRP3 inflammasome activation, caspase-1 (CASP1) cleavage, and proinflammatory interleukin 1 beta (IL1B) maturation. Gm15441-null mice were developed and shown to be more susceptible to NLRP3 inflammasome activation and to exhibit elevated CASP1 and IL1B cleavage in response to metabolic and inflammatory stimuli. These findings provide evidence for a novel mechanism by which PPARA attenuates hepatic inflammasome activation in response to metabolic stress through lncRNA Gm15441 induction. 4Txnip expression in vivo. Gm15441 LSL mice were treated with a PPARA agonist or fasted to assess how loss of Gm15441 impacts hepatic inflammasome activation in response to both pharmacological and physiologically-induced metabolic stress. TXNIP protein, caspase-1 (CASP1) levels, and interleukin 1 beta (IL1B) cleavage were elevated in Gm15441 LSL mice and were further increased by PPARA activation, indicating that this lncRNA plays a major role in attenuating inflammasome activation. Thus, hepatic PPARA directly regulates the lncRNA Gm15441, which in turn suppresses Txnip expression, which attenuates NLRP3 inflammasome activation during periods of metabolic stress. These studies revealed a novel regulatory mechanism supporting the beneficial effects of fasting, namely, reduced inflammation. Results LncRNA regulation by PPARA is highly tissue-specificTo assess the regulation of lncRNAs by PPARA, RNA-seq was performed using total liver RNA isolated from Ppara +/+ mice, both with and without dietary exposure to the PPARA agonist WY-14643. A lncRNA discovery pipeline was implemented that identified 15,558 liver-expressed lncRNA genes. Of these, 13,343 were intergenic, 1,966 were antisense, and 249 were intragenic lncRNAs. 44% of the 15,558 liver-expressed lncRNAs are considered novel (Melia and Waxman, 2019). Differential gene expression analysis revealed that a total of 1,735 RefSeq genes and 442 liver-expressed lncRNA genes responded to treatment with WY-14643 at an expression foldchange >2 at FDR<0.05, with 968 RefSeq genes and 245 lncRNA genes upregulated, and 767

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Dysregulation of a long noncoding RNA reduces leptin leading to a leptin-responsive form of obesity

Cited by 92 publications

References 43 publications

A non-canonical-PPARγ/RXRα-binding sequence regulates leptin expression in response to changes in adipose tissue mass

A non-canonical-PPARγ/RXRα-binding sequence regulates leptin expression in response to changes in adipose tissue mass

AGO HITS-CLIP in Adipose Tissue Reveals miR-29 as a Post-Transcriptional Regulator of Leptin

Long non-coding RNA Gm15441 attenuates hepatic inflammasome activation in response to metabolic stress

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