Regulation of adipogenesis and fat storage by IRX3 and IRX5

Obesity is a highly prevalent disease underlying several chronic diseases including Type 2 diabetes (T2D) and cardiovascular diseases (CVDs). Thus, increasing levels of obesity is associated with a series of co-morbidities and elevated risk of premature death. Obesity results from a chronic positive energy balance, causing white adipose tissue dysfunction, which in turn promotes dyslipidemia, systemic lipotoxicity and insulin resistance, eventually leading to ectopic fat accumulation and chronic diseases in multiple organs, including the heart, liver, arteries and kidneys. Unlike white adipocytes, beige fat cells are capable of disposing of excess energy by heat dissipation, thus protecting against obesity-related disease. Recently, two developmental transcription factors, IRX3 and IRX5, were shown to inhibit beige adipogenesis via an obesity associated risk genotype-dependent activation in preadipocytes. The aim of this study was therefore to investigate whether reducing IRX3 or IRX5 expression in adipose tissue offers protection from obesity, and if so, by which mechanisms. In paper I, we randomized wild type (WT) and Irx5 knock-out (KO) mice to a control or high-fat diet, and measured body weight, fat mass and global gene expression in adipose tissue. We found Irx5-KO mice to be lean and completely protected from diet-induced obesity. This was found to be partially attributable to increased mitochondrial respiration and thermogenesis due to reduction of Irx5 and App specifically in adipocytes. In papers II and III, we investigated the role of Irx3 in transcriptional regulation of adipogenesis, using WT and CRISPR-Cas9 mediated KO of Irx3 in preadipocytes, followed by RNA-, ATAC- and ChIP-sequencing. We found Irx3 to be critical for adipogenic identity and the ability of precursor cells to differentiate into mature adipocytes. Moreover, this lineage control was found to be mediated by direct transcriptional regulation of chromatin remodeling factors by Irx3. In conclusion, genetic repression of Irx3 or Irx5 offers strong protection against obesity, and reduces adipose tissue mass partially by increasing thermogenesis and improving mitochondrial respiration in existing adipocytes, and partially by preventing the formation of new adipocytes. This work has implications for identifying patients with genetic predisposition to obesity, who could benefit from potential therapeutic intervention targeting IRX3 or IRX5

Metabolic and Epigenetic Regulation by Estrogen in Adipocytes

Sex hormones contribute to differences between males and females in body fat distribution and associated disease risk. Higher concentrations of estrogens are associated with a more gynoid body shape and with more fat storage on hips and thighs rather than in visceral depots. Estrogen-mediated protection against visceral adiposity is shown in post-menopausal women with lower levels of estrogens and the reduction in central body fat observed after treatment with hormone-replacement therapy. Estrogen exerts its physiological effects via the estrogen receptors (ERα, ERβ and GPR30) in target cells, including adipocytes. Studies in mice indicate that estrogen protects against adipose inflammation and fibrosis also before the onset of obesity. The mechanisms involved in estrogen-dependent body fat distribution are incompletely understood, but involve, e.g., increased mTOR signaling and suppression of autophagy and adipogenesis/lipid storage. Estrogen plays a key role in epigenetic regulation of adipogenic genes by interacting with enzymes that remodel DNA methylation and histone tail post-translational modifications. However, more studies are needed to map the differential epigenetic effects of ER in different adipocyte subtypes, including those in subcutaneous and visceral adipose tissues. We here review recent discoveries of ER-mediated transcriptional and epigenetic regulation in adipocytes, which may explain sexual dimorphisms in body fat distribution and obesity-related disease risk.publishedVersio

COL6A3 expression in adipose tissue cells is associated with levels of the homeobox transcription factor PRRX1

Fibrillar collagen COL6α3 in adipose tissue has been associated with obesity, inflammation, insulin resistance and cancer. We here aimed to identify novel transcriptional regulators of COL6A3 expression. Based on a transcriptome dataset of adipose tissue, we identified strong correlations for 56 genes with COL6A3 mRNA, including targets of TGF-β/SMAD signaling. Among the identified candidates, the homeobox transcription factor PRRX1 showed a particularly striking co-expression with COL6A3, validated across several different cohorts, including patients with extreme obesity, insulin sensitive and resistant obesity (subcutaneous and omental), after profound fat loss (subcutaneous), and lean controls (subcutaneous). In human and mouse adipose cells, PRRX1 knockdown reduced COL6A3 mRNA and PRRX1 overexpression transactivated a reporter construct with the endogenous human COL6A3 promoter. Stable PRRX1 overexpression in 3T3-L1 cells induced Col6a3 mRNA threefold specifically after adipogenic induction, whereas TGF-β1 treatment upregulated Col6a3 mRNA also in the preadipocyte state. Interestingly, pro-inflammatory stimulus (i.e., TNF-α treatment) decreased PRRX1-mediated Col6a3 transactivation and mRNA expression, supporting a role for this mechanism in the regulation of adipose tissue inflammation. In conclusion, we identified the homeobox factor PRRX1 as a novel transcriptional regulator associated with COL6A3 expression, providing new insight into the regulatory mechanisms of altered adipose tissue function in obesity and insulin resistance.publishedVersio

Perceived health status: is obesity perceived as a risk factor and disease?

One might expect that a perception of obesity being a risk factor and disease, contributes to effective obesity prevention and management strategies. However, obesity rates continue to increase worldwide. The question arises whether obesity is truly perceived as a risk factor and a disease. This paper aims at describing perception of obesity as risk factor and disease among individuals seeking care, individuals not seeking care, the society, and different professionals having a role in the field of obesity. The paper is a reflection of the lecture on the topic that was given at the EASO's New Investigators United's Summer School 2016 in Portugal and the discussion with the new investigators and other senior speakers. Individual obese patients seeking help are very much aware of obesity being a risk factor and disease, but perceptions regarding obesity seem to be flawed among those who do not seek help for obesity. Also, misperceptions regarding obesity play a role at different levels, including society, different political levels, the fields of health care and social work, prevention organizations, and the food and marketing industry. The food and marketing industry has an enormous role in changing perceptions by the society and policy makers. Obesity rates will continue to increase as long as individuals, the society, and professionals at different levels have false interpretations of the severity of obesity. Severe action is needed against those who are playing a role in maintaining false perceptions of obesity as a risk factor and disease.- (undefined

Rapid climate changes during the Lateglacial and the early Holocene as seen from plant community dynamics in the Polar Urals, Russia

A detailed, well-dated record of pollen and sedimentary ancient DNA (sedaDNA) for the period 15 000–9500 cal a bp describes changes at Lake Bolshoye Shchuchye in the Polar Ural Mountains, located far east of the classical Lateglacial sites in western Europe. Arctic tundra rapidly changed to lusher vegetation, possibly including both dwarf (Betula nana) and tree birch (B. pubescens), dated in our record to take place 14 565 cal a bp, coincident with the onset of the Bølling in western Europe; this was paralleled by increased summer temperatures. A striking feature is an early decline in Betula pollen and sedaDNA reads 300 years before the onset of the Younger Dryas (YD) in western Europe. Given the solid site chronology, this could indicate that the YD cooling started in Siberia and propagated westwards, or that the vegetation reacted to the inter-Allerød cooling at 13 100 cal a bp and did not recover during the late Allerød. During the YD, increases in steppe taxa such as Artemisia and Chenopodiaceae suggest drier conditions. At the onset of the Holocene, the vegetation around the lake reacted fast to the warmer conditions, as seen in the increase of arboreal taxa, especially Betula, and a decrease in herbs such as Artemisia and Cyperaceae.publishedVersio

A 24,000-year ancient DNA and pollen record from the Polar Urals reveals temporal dynamics of arctic and boreal plant communities

A 24,000-year record of plant community dynamics, based on pollen and ancient DNA from the sediments (sedaDNA) of Lake Bolshoye Shchuchye in the Polar Ural Mountains, provides detailed information on the flora of the Last Glacial Maximum (LGM) and also changes in plant community composition and dominance. It greatly improves on incomplete records from short and fragmented stratigraphic sequences found in exposed sedimentary sections in the western Russian Arctic. In total, 162 plant taxa were detected by sedaDNA and 115 by pollen analysis. Several shifts in dominance between and within plant functional groups occurred over the studied period, but most taxa appear to have survived in situ. A diverse arctic-alpine herb flora characterised the interval ca. 24,000–17,000 cal years BP and persisted into the Holocene. Around 17,000 cal years BP, sedges (e.g. Carex) and bryophytes (e.g. Bryum, Aulacomnium) increased. The establishment of shrub-tundra communities of Dryas and Vaccinium sp., with potentially some Betula pubescens trees (influx ∼290 grains cm2 year−1), followed at ca. 15,000 cal years BP. Forest taxa such as Picea and ferns (e.g. Dryopteris fragrans, Gymnocarpium dryopteris) established near the lake from ca. 10,000 cal years BP, followed by the establishment of Larix trees from ca. 9000 cal years BP. Picea began to decline from ca. 7000 cal years BP. A complete withdrawal of forest tree taxa occurred by ca. 4000 cal years BP, presumably due to decreasing growing-season temperatures, allowing the expansion of dwarf-shrub tundra and a diverse herb community similar to the present-day vegetation mosaic. Contrary to some earlier comparative studies, sedaDNA and pollen from Lake Bolshoye Shchuchye showed high similarity in the timing of compositional changes and the occurrence of key plant taxa. The sedaDNA record revealed several features that the pollen stratigraphy and earlier palaeorecords in the region failed to detect; a sustained, long-term increase in floristic richness since the LGM until the early Holocene, turnover in grass and forb genera over the Pleistocene-Holocene transition, persistence of a diverse arctic-alpine flora over the late Quaternary, and a variable bryophyte flora through time. As pollen records are often limited by taxonomic resolution, differential productivity and dispersal, sedaDNA can provide improved estimates of floristic richness and is better able to distinguish between different plant assemblages. However, pollen remains superior at providing quantitative estimates of plant abundance changes and detecting several diverse groups (e.g. Poaceae, Cyperaceae, Asteraceae) which may be underreported in the sedaDNA. Joint use of the two proxies provided unprecedented floristic detail of past plant communities and helped to distinguish between long-distance transport of pollen and local presence, particularly for woody plant taxa

Role of the Neutral Amino Acid Transporter SLC7A10 in Adipocyte Lipid Storage, Obesity and Insulin Resistance

Elucidation of mechanisms that govern lipid storage, oxidative stress, and insulin resistance may lead to improved therapeutic options for type 2 diabetes and other obesity-related diseases. Here, we find that adipose expression of the small neutral amino acid transporter SLC7A10, also known as alanine-serine-cysteine transporter-1 (ASC-1), shows strong inverse correlates with visceral adiposity, insulin resistance, and adipocyte hypertrophy across multiple cohorts. Concordantly, loss of Slc7a10 function in zebrafish in vivo accelerates diet-induced body weight gain and adipocyte enlargement. Mechanistically, SLC7A10 inhibition in human and murine adipocytes decreases adipocyte serine uptake and total glutathione levels and promotes reactive oxygen species (ROS) generation. Conversely, SLC7A10 overexpression decreases ROS generation and increases mitochondrial respiratory capacity. RNA sequencing revealed consistent changes in gene expression between human adipocytes and zebrafish visceral adipose tissue following loss of SLC7A10, e.g., upregulation of SCD (lipid storage) and downregulation of CPT1A (lipid oxidation). Interestingly, ROS scavenger reduced lipid accumulation and attenuated the lipid-storing effect of SLC7A10 inhibition. These data uncover adipocyte SLC7A10 as a novel important regulator of adipocyte resilience to nutrient and oxidative stress, in part by enhancing glutathione levels and mitochondrial respiration, conducive to decreased ROS generation, lipid accumulation, adipocyte hypertrophy, insulin resistance, and type 2 diabetes.acceptedVersio

Regulation of adipogenesis and fat storage by IRX3 and IRX5

Obesity is a highly prevalent disease underlying several chronic diseases including Type 2 diabetes (T2D) and cardiovascular diseases (CVDs). Thus, increasing levels of obesity is associated with a series of co-morbidities and elevated risk of premature death. Obesity results from a chronic positive energy balance, causing white adipose tissue dysfunction, which in turn promotes dyslipidemia, systemic lipotoxicity and insulin resistance, eventually leading to ectopic fat accumulation and chronic diseases in multiple organs, including the heart, liver, arteries and kidneys. Unlike white adipocytes, beige fat cells are capable of disposing of excess energy by heat dissipation, thus protecting against obesity-related disease. Recently, two developmental transcription factors, IRX3 and IRX5, were shown to inhibit beige adipogenesis via an obesity associated risk genotype-dependent activation in preadipocytes. The aim of this study was therefore to investigate whether reducing IRX3 or IRX5 expression in adipose tissue offers protection from obesity, and if so, by which mechanisms. In paper I, we randomized wild type (WT) and Irx5 knock-out (KO) mice to a control or high-fat diet, and measured body weight, fat mass and global gene expression in adipose tissue. We found Irx5-KO mice to be lean and completely protected from diet-induced obesity. This was found to be partially attributable to increased mitochondrial respiration and thermogenesis due to reduction of Irx5 and App specifically in adipocytes. In papers II and III, we investigated the role of Irx3 in transcriptional regulation of adipogenesis, using WT and CRISPR-Cas9 mediated KO of Irx3 in preadipocytes, followed by RNA-, ATAC- and ChIP-sequencing. We found Irx3 to be critical for adipogenic identity and the ability of precursor cells to differentiate into mature adipocytes. Moreover, this lineage control was found to be mediated by direct transcriptional regulation of chromatin remodeling factors by Irx3. In conclusion, genetic repression of Irx3 or Irx5 offers strong protection against obesity, and reduces adipose tissue mass partially by increasing thermogenesis and improving mitochondrial respiration in existing adipocytes, and partially by preventing the formation of new adipocytes. This work has implications for identifying patients with genetic predisposition to obesity, who could benefit from potential therapeutic intervention targeting IRX3 or IRX5

Metformin inhibits hepatocellular glucose, lipid and cholesterol biosynthetic pathways by transcriptionally suppressing steroid receptor coactivator 2 (SRC-2)

The ability of the anti-diabetic drug metformin to inhibit anabolic processes including gluconeogenesis and lipogenesis is partly attributable to activation of the AMP-activated protein kinase (AMPK) pathway. The p160 steroid receptor coactivator 2 (SRC-2) is a key regulator of cellular metabolism and drives expression of the gluconeogenic enzyme glucose-6-phosphatase (G6Pc). Here, we uncovered a role for SRC-2 in the metabolic reprogramming imposed by metformin. In FaO cells, metformin dose-dependently reduced mRNA expression of SRC-2. Microarray analysis of metformin- treated cells revealed an overrepresentation of downregulated genes involved in biosynthesis of lipids and cholesterol. Several metformin-regulated genes including fatty acid synthase (FASN) were validated as transcriptional targets of SRC-2 with promoters characterized by sterol regulatory element (SRE) binding protein (SREBP) recognition sequences. Transactivation assays of the FASN promoter confirmed that SRC-2 is a coactivator of SREBP-1. By suppressing SRC-2 at the transcriptional level, metformin impeded recruitment of SRC-2 and RNA polymerase II to the G6Pc promoter and to SREs of mutual SRC-2/SREBP-1 target gene promoters. Hepatocellular fat accretion was reduced by metformin or knock-down of both SRC-2 and SREBP-1. Accordingly we propose that metformin inhibits glucose and lipid biosynthesis partly by downregulating SRC-2 gene expression

Metabolic and Epigenetic Regulation by Estrogen in Adipocytes

Sex hormones contribute to differences between males and females in body fat distribution and associated disease risk. Higher concentrations of estrogens are associated with a more gynoid body shape and with more fat storage on hips and thighs rather than in visceral depots. Estrogen-mediated protection against visceral adiposity is shown in post-menopausal women with lower levels of estrogens and the reduction in central body fat observed after treatment with hormone-replacement therapy. Estrogen exerts its physiological effects via the estrogen receptors (ERα, ERβ and GPR30) in target cells, including adipocytes. Studies in mice indicate that estrogen protects against adipose inflammation and fibrosis also before the onset of obesity. The mechanisms involved in estrogen-dependent body fat distribution are incompletely understood, but involve, e.g., increased mTOR signaling and suppression of autophagy and adipogenesis/lipid storage. Estrogen plays a key role in epigenetic regulation of adipogenic genes by interacting with enzymes that remodel DNA methylation and histone tail post-translational modifications. However, more studies are needed to map the differential epigenetic effects of ER in different adipocyte subtypes, including those in subcutaneous and visceral adipose tissues. We here review recent discoveries of ER-mediated transcriptional and epigenetic regulation in adipocytes, which may explain sexual dimorphisms in body fat distribution and obesity-related disease risk