A single day of high-fat diet feeding induces lipid accumulation and insulin resistance in brown adipose tissue in mice
Kuipers, Eline N. ; Held, Ntsiki M. ; Het Panhuis, Wietse In; Modder, Melanie ; Ruppert, Philip M.M. ; Kersten, Sander ; Kooijman, Sander ; Guigas, Bruno ; Houtkooper, Riekelt H. ; Rensen, Patrick C.N. ; Boon, Mariëtte R. - \ 2019
American Journal of Physiology. Endocrinology and Metabolism 317 (2019)5. - ISSN 0193-1849 - p. E820 - E830.
brown adipose tissue - high-fat diet - lipid accumulation - macrophage - mitochondrial dynamics
Brown adipose tissue (BAT) catabolizes glucose and fatty acids to produce heat and thereby contributes to energy expenditure. Long-term high-fat diet (HFD) feeding results in so-called 'whitening' of BAT characterized by increased lipid deposition, mitochondrial dysfunction, and reduced fat oxidation. The aim of the current study was to unravel the rate and related mechanisms by which HFD induces BAT whitening and insulin resistance. Wild-type mice were fed a HFD for 0, 1, 3, or 7 days. Within 1 day of HFD, BAT weight and lipid content were increased. HFD also immediately reduced insulin-stimulated glucose uptake by BAT, indicating rapid induction of insulin resistance. This was accompanied by a tendency toward a reduced uptake of triglyceride-derived fatty acids by BAT. Mitochondrial mass and Ucp1 expression were unaltered, whereas after 3 days of HFD, markers of mitochondrial dynamics suggested induction of a more fused mitochondrial network. Additionally, HFD also increased macrophage markers in BAT after 3 days of HFD. Counterintuitively, the switch to HFD was accompanied by an acute rise in core body temperature. We showed that a single day of HFD feeding is sufficient to induce the first signs of whitening and insulin resistance in BAT, which reduces the uptake of glucose and triglyceride-derived fatty acids. BAT whitening and insulin resistance are likely sustained by reduced mitochondrial oxidation due to changes in mitochondrial dynamics and macrophage infiltration, respectively. Likely, the switch to HFD swiftly induces thermogenesis in other metabolic organs, which allows attenuation of BAT thermogenesis.
Natural genetic variation in C. elegans identified genomic loci controlling metabolite levels
Gao, Arwen W. ; Sterken, Mark G. ; De Bos, Jelmi Uit; Creij, Jelle van; Kamble, Rashmi ; Snoek, Basten L. ; Kammenga, Jan E. ; Houtkooper, Riekelt H. - \ 2018
Genome Research 28 (2018)8. - ISSN 1088-9051 - p. 1296 - 1308.
Metabolic homeostasis is sustained by complex biological networks that respond to nutrient availability. Genetic and environmental factors may disrupt this equilibrium, leading to metabolic disorders, including obesity and type 2 diabetes. To identify the genetic factors controlling metabolism, we performed quantitative genetic analysis using a population of 199 recombinant inbred lines (RILs) in the nematode Caenorhabditis elegans. We focused on the genomic regions that control metabolite levels by measuring fatty acid (FA) and amino acid (AA) composition in the RILs using targeted metabolomics. The genetically diverse RILs showed a large variation in their FA andAAlevels with a heritability ranging from 32% to 82%. We detected strongly co-correlated metabolite clusters and 36 significant metabolite QTL (mQTL). We focused on mQTL displaying highly significant linkage and heritability, including an mQTL for the FA C14:1 on Chromosome I, and another mQTL for the FA C18:2 on Chromosome IV. Using introgression lines (ILs), we were able to narrow down both mQTL to a 1.4-Mbp and a 3.6-Mbp region, respectively. RNAi-based screening focusing on the Chromosome I mQTL identified several candidate genes for the C14:1 mQTL, including lagr-1, Y87G2A.2, nhr-265, nhr-276, and nhr-81. Overall, this systems
approach provides us with a powerful platform to study the genetic basis of C. elegans metabolism. Furthermore, it allows us to investigate interventions such as nutrients and stresses that maintain or disturb the regulatory network controlling metabolic homeostasis, and identify gene-by-environment interactions.
|Natural genetic variation in C. elegans identified genomic loci controlling metabolite levels
Sterken, M.G. ; Gao, Arwen W. ; Bos, Jelmi uit de; Creij, J.W. van; Kamble, Rashmi ; Snoek, L.B. ; Kammenga, J.E. ; Houtkooper, Riekelt H. - \ 2018
Metabolic homeostasis is sustained by complex biological networks responding to nutrient availability. Genetic and/or environmental perturbations can lead to metabolic disorders, including obesity and type-2 diabetes. Model organisms are particularly suited to study the interactions between genetic and environmental factors. Thus far, metabolism in C. elegans was often studied at the transcriptional level opposed to the metabolite level. Using a recently developed metabolomics platform, we were able to measure metabolites in C. elegans on a large scale. Here, we aimed to identify the genetic factors controlling metabolism using the nematode Caenorhabditis elegans.
We used a quantitative genetic approach with a C. elegans population consisting of 199 recombinant inbred lines (RILs). We measured fatty acid (FA) and amino acid (AA) composition in the RILs using targeted metabolomics. We were able to measure the metabolite levels of 56 metabolites. Subsequently, we determined transgression and heritabilities for these metabolites. We found large variation in metabolites levels and for 18 metabolites significant transgression was found, these metabolites were predominantly FAs. The heritability was significant for 51 metabolites and ranged between 32 to 82%. Using a single marker model, we found 36 significant metabolite quantitative trait loci (mQTL). Additionally, a full two-marker screen revealed interacting loci for 6 metabolites. Using introgression lines (ILs) we verified the mQTL for two FA’s, C14:1 and C18:2 mapping to chromosome I and IV respectively. We narrowed down both mQTL to a 1.4 Mbp and a 3.6 Mbp region, respectively. Focussing on the chromosome I mQTL we conducted a prioritized candidate screening, revealing several candidate genes that could affect C14:1 levels across the RILs. By RNAi based knock-down in the N2 strain we could verify that five candidate genes affect C14:1 levels: lagr-1, Y87G2A.2, nhr-265, nhr-276, and nhr-81.
In conclusion, genetic variation affecting metabolite levels in C. elegans proofed to be extensive. Natural variation in C. elegans can play an important role to dissect the mechanisms underlying the complex processes of metabolism in a natural and unbiased manner and allow us to identify factors important for gene-by-environment interactions. Therefore, our study provides the basis to investigate additional interventions, such as nutrients and stresses that maintain or disturb the regulatory network controlling metabolic homeostasis.
Forward and reverse genetics approaches to uncover metabolic aging pathways in Caenorhabditis elegans
Gao, Arwen W. ; Bos, Jelmi uit de; Sterken, Mark G. ; Kammenga, Jan E. ; Smith, Reuben L. ; Houtkooper, Riekelt H. - \ 2018
Biochimica et Biophysica Acta. Molecular Basis of Disease 1864 (2018)9A. - ISSN 0925-4439 - p. 2697 - 2706.
The biological mechanisms of aging have been studied in depth and prominent findings in this field promote the development of new therapies for age-associated disorders. Various model organisms are used for research on aging; among these, the nematode Caenorhabditis elegans has been widely used and has provided valuable knowledge in determining the regulatory mechanisms driving the aging process. Many genes involved in lifespan regulation are associated with metabolic pathways and are influenced by genetic and environmental factors. In line with this, C. elegans provides a promising platform to study such gene by environment interactions, in either a reverse or forward genetics approach. In this review, we discuss longevity mechanisms related to metabolic networks that have been discovered in C. elegans. We also highlight the use of wild populations to study the complex genetic basis of natural variation for quantitative traits that mediate longevity.
The gut microbiota as a modulator of innate immunity during melioidosis
Lankelma, Jacqueline M. ; Birnie, Emma ; Weehuizen, Tassili A.F. ; Scicluna, Brendon P. ; Belzer, Clara ; Houtkooper, Riekelt H. ; Roelofs, Joris J.T.H. ; Vos, Alex F. de; Poll, Tom van der; Budding, Andries E. ; Wiersinga, W.J. - \ 2017
PLoS Neglected Tropical Diseases 11 (2017)4. - ISSN 1935-2727
Background: Melioidosis, caused by the Gram-negative bacterium Burkholderia pseudomallei, is an emerging cause of pneumonia-derived sepsis in the tropics. The gut microbiota supports local mucosal immunity and is increasingly recognized as a protective mediator in host defenses against systemic infection. Here, we aimed to characterize the composition and function of the intestinal microbiota during experimental melioidosis. Methodology/Principal findings: C57BL/6 mice were infected intranasally with B. pseudomallei and sacrificed at different time points to assess bacterial loads and inflammation. In selected experiments, the gut microbiota was disrupted with broad-spectrum antibiotics prior to inoculation. Fecal bacterial composition was analyzed by means of IS-pro, a 16S-23S interspacer region-based profiling method. A marked shift in fecal bacterial composition was seen in all mice during systemic B. pseudomallei infection with a strong increase in Proteobacteria and decrease in Actinobacteria, with an increase in bacterial diversity. We found enhanced early dissemination of B. pseudomallei and systemic inflammation during experimental melioidosis in microbiota-disrupted mice compared with controls. Whole-genome transcriptional profiling of the lung identified several genes that were differentially expressed between mice with a normal or disrupted intestinal microbiota. Genes involved in acute phase signaling, including macrophage-related signaling pathways were significantly elevated in microbiota disrupted mice. Compared with controls, alveolar macrophages derived from antibiotic pretreated mice showed a diminished capacity to phagocytose B. pseudomallei. This might in part explain the observed protective effect of the gut microbiota in the host defense against pneumonia-derived melioidosis. Conclusions/Significance: Taken together, these data identify the gut microbiota as a potential modulator of innate immunity during B. pseudomallei infection.
Microbial stimulation of different Toll-like receptor signalling pathways induces diverse metabolic programmes in human monocytes
Lachmandas, Ekta ; Boutens, Lily ; Ratter, Jacqueline M. ; Hijmans, Anneke ; Hooiveld, Guido J. ; Joosten, Leo A.B. ; Rodenburg, Richard J. ; Fransen, Jack A.M. ; Houtkooper, Riekelt H. ; Crevel, R. van; Netea, Mihai G. ; Stienstra, R. - \ 2016
Nature Microbiology 2 (2016). - ISSN 2058-5276
Microbial stimuli such as lipopolysaccharide (LPS) induce robust metabolic rewiring in immune cells known as the Warburg effect. It is unknown whether this increase in glycolysis and decrease in oxidative phosphorylation (OXPHOS) is a general characteristic of monocytes that have encountered a pathogen. Using CD14+ monocytes from healthy donors, we demonstrated that most microbial stimuli increased glycolysis, but that only stimulation of Toll-like receptor (TLR) 4 with LPS led to a decrease in OXPHOS. Instead, activation of other TLRs, such as TLR2 activation by Pam3CysSK4 (P3C), increased oxygen consumption and mitochondrial enzyme activity. Transcriptome and metabolome analysis of monocytes stimulated with P3C versus LPS confirmed the divergent metabolic responses between both stimuli, and revealed significant differences in the tricarboxylic acid cycle, OXPHOS and lipid metabolism pathways following stimulation of monocytes with P3C versus LPS. At a functional level, pharmacological inhibition of complex I of the mitochondrial electron transport chain diminished cytokine production and phagocytosis in P3C- but not LPS-stimulated monocytes. Thus, unlike LPS, complex microbial stimuli and the TLR2 ligand P3C induce a specific pattern of metabolic rewiring that involves upregulation of both glycolysis and OXPHOS, which enables activation of host defence mechanisms such as cytokine production and phagocytosis.
Mannose-Binding Lectin Is Required for the Effective Clearance of Apoptotic Cells by Adipose Tissue Macrophages During Obesity
Stienstra, R. ; Dijk, W. ; Beek, L. van; Jansen, H. ; Heemskerk, M. ; Houtkooper, R.H. ; Denis, S. ; Harmelen, V. van; Willems van Dijk, K. ; Tack, C.J. ; Kersten, A.H. - \ 2014
Diabetes 63 (2014)12. - ISSN 0012-1797 - p. 4143 - 4153.
insulin-resistance - cytokine secretion - hepatic steatosis - dendritic cells - deficient mice - tnf-alpha - innate - protein - inflammation - adipocytes
Obesity is accompanied by the presence of chronic low-grade inflammation manifested by infiltration of macrophages into adipose tissue. Mannose-binding lectin (MBL), a soluble mediator of innate immunity, promotes phagocytosis and alters macrophage function. To assess the function of MBL in the development of obesity, we studied wild-type and MBL-/- mice rendered obese using a high-fat diet (HFD). Whereas no gross morphological differences were observed in liver, an HFD provoked distinct changes in the adipose tissue morphology of MBL-/- mice. In parallel with increased adipocyte size, MBL-/- mice displayed an increased influx of macrophages into adipose tissue. Macrophages were polarized toward an alternatively activated phenotype known to modulate apoptotic cell clearance. MBL deficiency also significantly increased the number of apoptotic cells in adipose tissue. Consistent with these observations, recombinant MBL enhanced phagocytic capacity of the stromal vascular fraction isolated from adipose tissue and modulated uptake of apoptotic adipocytes by macrophages. Despite changes in macrophage abundance and polarity, the absence of MBL did not affect systemic insulin resistance. Finally, in humans, lower levels of circulating MBL were accompanied by enhanced macrophage influx in subcutaneous adipose tissue. We propose a novel role for MBL in the recognition and clearance of apoptotic adipocytes during obesity.
Calorie restriction-like effects of 30 days of resveratrol supplementation on energy metabolism and metabolic profile in obese humans
Timmers, Silvie ; Konings, Ellen ; Bilet, Lena ; Houtkooper, Riekelt H. ; Weijer, Tineke van de; Hoeks, Joris ; Krieken, Sophie van der; Ryu, Dongryeol ; Kersten, Sander ; Moonen-Kornips, Esther ; Goossens, Gijs H. ; Hesselink, Matthijs K. ; Kunz, Iris ; Schrauwen-Hinderling, Vera B. ; Blaak, Ellen E. ; Auwerx, Johan ; Schrauwen, Patrick - \ 2011
Homo sapiens - GSE32357 - PRJNA14724 - Homo sapiens - GSE32357
Resveratrol is a naturally occurring compound that profoundly affects energy metabolism and mitochondrial function and serves as a calorie restriction mimetic, at least in animal models of obesity. Here we treated 10 healthy, obese men with placebo and 150 mg/day resveratrol in a randomized double-blind cross-over study for 30 days. Resveratrol supplementation significantly reduced sleeping- and resting metabolic rate. In muscle, resveratrol activated AMPK, increased SIRT1 and PGC-1alpha protein levels, increased citrate synthase activity, and improved muscle mitochondrial respiration on a fatty acid-derived substrate. Furthermore, resveratrol elevated intramyocellular lipid levels, and decreased intrahepatic lipid content, circulating glucose, triglycerides, alanine-aminotransferase, and inflammation markers. Systolic blood pressure dropped and HOMA index improved after resveratrol. In the postprandial state, adipose tissue lipolysis and plasma fatty acid and glycerol decreased. In conclusion, we demonstrate that 30 days of resveratrol supplementation induces profound metabolic changes in obese subjects, mimicking the effects of calorie restriction.
Calorie Restriction-like Effects of 30 Days of Resveratrol Supplementation on Energy Metabolism and Metabolic Profile in Obese Humans
Timmers, S. ; Konings, E. ; Bilet, L. ; Houtkooper, R.H. ; Weijer, T. van de; Goossens, G.H. ; Hoeks, J. ; Krieken, S. van der; Ryu, D. ; Kersten, A.H. ; Moonen-Kornips, E. ; Hesselink, M.K.C. ; Kunz, I. ; Schrauwen-Hinderling, V.B. ; Blaak, E.E. ; Auwerx, J. ; Schrauwen, P. - \ 2011
Cell Metabolism 14 (2011)5. - ISSN 1550-4131 - p. 612 - 622.
beta-cell function - life-span - insulin sensitivity - mitochondrial-function - skeletal-muscle - saccharomyces-cerevisiae - sirt1 activation - lipid-metabolism - fat oxidation - exercise
Resveratrol is a natural compound that affects energy metabolism and mitochondrial function and serves as a calorie restriction mimetic, at least in animal models of obesity. Here, we treated 11 healthy, obese men with placebo and 150 mg/day resveratrol (resVida) in a randomized double-blind crossover study for 30 days. Resveratrol significantly reduced sleeping and resting metabolic rate. In muscle, resveratrol activated AMPK, increased SIRT1 and PGC-1 alpha protein levels, increased citrate synthase activity without change in mitochondrial content, and improved muscle mitochondrial respiration on a fatty acid-derived substrate. Furthermore, resveratrol elevated intramyocellular lipid levels and decreased intrahepatic lipid content, circulating glucose, triglycerides, alanine-aminotransferase, and inflammation markers. Systolic blood pressure dropped and HOMA index improved after resveratrol. In the postprandial state, adipose tissue lipolysis and plasma fatty acid and glycerol decreased. In conclusion, we demonstrate that 30 days of resveratrol supplementation induces metabolic changes in obese humans, mimicking the effects of calorie restriction.