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01-02-2013 | Article

Functional relevance of genes implicated by obesity genome-wide association study signals for human adipocyte biology

Authors: F. Bernhard, K. Landgraf, N. Klöting, A. Berthold, P. Büttner, D. Friebe, W. Kiess, P. Kovacs, M. Blüher, A. Körner

Published in: Diabetologia | Issue 2/2013

Abstract

Aims/hypothesis

Genome-wide association studies (GWAS) have identified numerous single-nucleotide polymorphisms associated with obesity, consequently implying a role in adipocyte biology for many closely residing genes. We investigated the functional relevance of such genes in human adipocytes.

Methods

We selected eight genes (BDNF, MAF, MTCH2, NEGR1, NPC1, PTER, SH2B1 and TMEM18) from obesity GWAS and analysed their effect in human adipogenesis using small interfering (si)RNA-mediated knockdown, their regulation by metabolic agents in adipocytes and pre-adipocytes, and gene expression in paired samples of human fat biopsies (68 non-obese, 165 obese) by quantitative real-time PCR.

Results

We show a two- to threefold upregulation of MAF, MTCH2 and NEGR1 and a two- to fourfold downregulation of BDNF and PTER during adipogenesis. Knockdown of BDNF (mean ± SEM; 83.8 ± 4.7% of control; p = 0.0002), MTCH2 (72.7 ± 9.5%; p = 0.0006), NEGR1 (70.2 ± 5.7%; p < 0.0001) and TMEM18 (70.8 ± 6.1%; p < 0.0001) significantly inhibited adipocyte maturation, while knockdown of the other proteins had no effect. Insulin slightly induced MAF (1.65-fold; p = 0.0009) and MTCH2 (1.72-fold; p < 0.0001), while it suppressed BDNF (59.6%; p = 0.0009), NEGR1 (58.0%; p = 0.0085) and TMEM18 (69.3%; p = 0.0377) in adipocytes. The synthetic glucocorticoid dexamethasone suppressed MAF (45.7%; p = 0.0022), BDNF (66.6%; p = 0.0012) and TMEM18 (63.5%; p = 0.0181), but induced NEGR1 (3.2-fold; p = 0.0117) expression. Furthermore, MTCH2, NEGR1 and TMEM18 were differentially expressed in subcutaneous and visceral adipose tissue. TMEM18 expression was decreased in the adipose tissue of obese patients, and negatively correlated with anthropometric variables and adipocyte size.

Conclusions/interpretation

Our results imply a regulatory role for TMEM18, BDNF, MTCH2 and NEGR1 in adipocyte differentiation and biology. In addition, we show a variation of MAF expression during adipogenesis, while NPC1, PTER and SH2B1 were not regulated.

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Meyre D, Delplanque J, Chèvre JC et al (2009) Genome-wide association study for early-onset and morbid adult obesity identifies three new risk loci in European populations. Nat Genet 41:157–159PubMedCrossRef

Thorleifsson G, Walters GB, Gudbjartsson DF et al (2009) Genome-wide association yields new sequence variants at seven loci that associate with measures of obesity. Nat Genet 41:18–24PubMedCrossRef

Willer CJ, Speliotes EK, Loos RJ et al (2009) Six new loci associated with body mass index highlight a neuronal influence on body weight regulation. Nat Genet 41:25–34PubMedCrossRef

Cypess AM, Lehman S, Williams G et al (2009) Identification and importance of brown adipose tissue in adult humans. N Engl J Med 360:1509–1517PubMedCrossRef

Ichimura A, Hirasawa A, Poulain-Godefroy O et al (2012) Dysfunction of lipid sensor GPR120 leads to obesity in both mouse and human. Nature 483:350–354PubMedCrossRef

Greenblum S, Turnbaugh PJ, Borenstein E (2012) Metagenomic systems biology of the human gut microbiome reveals topological shifts associated with obesity and inflammatory bowel disease. Proc Natl Acad Sci USA 109:594–599PubMedCrossRef

Cristancho AG, Lazar MA (2011) Forming functional fat: a growing understanding of adipocyte differentiation. Nat Rev Mol Cell Biol 12:722–734PubMedCrossRef

Wabitsch M, Brenner RE, Melzner I et al (2001) Characterization of a human pre-adipocyte cell strain with high capacity for adipose differentiation. Int J Obes Relat Metab Disord 25:8–15PubMedCrossRef

Körner A, Wabitsch M, Seidel B et al (2005) Adiponectin expression in humans is dependent on differentiation of adipocytes and down-regulated by humoral serum components of high molecular weight. Biochem Biophys Res Commun 337:540–550PubMedCrossRef

10.

Gagnon A, Sorisky A (1998) The effect of glucose concentration on insulin-induced 3 T3-L1 adipose cell differentiation. Obes Res 6:157–163PubMed

11.

Smith PJ, Wise LS, Berkowitz R, Wan C, Rubin CS (1988) Insulin-like growth factor-I is an essential regulator of the differentiation of 3 T3-L1 adipocytes. J Biol Chem 263:9402–9408PubMed

12.

Zilberfarb V, Siquier K, Strosberg AD, Issad T (2001) Effect of dexamethasone on adipocyte differentiation markers and tumour necrosis factor-alpha expression in human PAZ6 cells. Diabetologia 44:377–386PubMedCrossRef

13.

Burns TW, Langley PE, Terry BE et al (1981) Pharmacological characterizations of adrenergic receptors in human adipocytes. J Clin Invest 67:467–475PubMedCrossRef

14.

Kloting N, Graham TE, Berndt J et al (2007) Serum retinol-binding protein is more highly expressed in visceral than in subcutaneous adipose tissue and is a marker of intra-abdominal fat mass. Cell Metab 6:79–87PubMedCrossRef

15.

Heard-Costa NL, Zillikens MC, Monda KL et al (2009) NRXN3 is a novel locus for waist circumference: a genome-wide association study from the CHARGE Consortium. PLoS Genet 5:e1000539PubMedCrossRef

16.

Lindgren CM, Heid IM, Randall JC et al (2009) Genome-wide association scan meta-analysis identifies three loci influencing adiposity and fat distribution. PLoS Genet 5:e1000508PubMedCrossRef

17.

Renström F, Payne F, Nordström A et al (2009) Replication and extension of genome-wide association study results for obesity in 4923 adults from northern Sweden. Hum Mol Genet 18:1489–1496PubMedCrossRef

18.

Friebe D, Löffler D, Schönberg M et al (2011) Impact of metabolic regulators on the expression of the obesity associated genes FTO and NAMPT in human pre-adipocytes and adipocytes. PLoS One 6:e19526PubMedCrossRef

19.

Boissel S, Reish O, Proulx K et al (2009) Loss-of-function mutation in the dioxygenase-encoding FTO gene causes severe growth retardation and multiple malformations. Am J Hum Genet 85:106–111PubMedCrossRef

20.

Fischer J, Koch L, Emmerling C et al (2009) Inactivation of the Fto gene protects from obesity. Nature 458:894–898PubMedCrossRef

21.

Mountjoy KG, Mortrud MT, Low MJ, Simerly RB, Cone RD (1994) Localization of the melanocortin-4 receptor (MC4-R) in neuroendocrine and autonomic control circuits in the brain. Mol Endocrinol 8:1298–1308PubMedCrossRef

22.

Huszar D, Lynch CA, Fairchild-Huntress V et al (1997) Targeted disruption of the melanocortin-4 receptor results in obesity in mice. Cell 88:131–141PubMedCrossRef

23.

Siersbaek R, Nielsen R, Mandrup S (2010) PPARgamma in adipocyte differentiation and metabolism—novel insights from genome-wide studies. FEBS Lett 584:3242–3249PubMedCrossRef

24.

Berndt J, Klöting N, Kralisch S et al (2005) Plasma visfatin concentrations and fat depot-specific mRNA expression in humans. Diabetes 54:2911–2916PubMedCrossRef

25.

Jurvansuu J, Zhao Y, Leung DSY et al (2008) Transmembrane protein 18 enhances the tropism of neural stem cells for glioma cells. Cancer Res 68:4614–4622PubMedCrossRef

26.

Scherag A, Dina C, Hinney A et al (2010) Two new loci for body-weight regulation identified in a joint analysis of genome-wide association studies for early-onset extreme obesity in French and German study groups. PLoS Genet 6:e1000916PubMedCrossRef

27.

Speliotes EK, Willer CJ, Berndt SI et al (2010) Association analyses of 249,796 individuals reveal 18 new loci associated with body mass index. Nat Genet 42:937–948PubMedCrossRef

28.

Nagahara AH, Tuszynski MH (2011) Potential therapeutic uses of BDNF in neurological and psychiatric disorders. Nat Rev Drug Discov 10:209–219PubMedCrossRef

29.

Cordeira J, Rios M (2011) Weighing in the role of BDNF in the central control of eating behavior. Mol Neurobiol 44:441–448PubMedCrossRef

30.

Xu B, Goulding EH, Zang K et al (2003) Brain-derived neurotrophic factor regulates energy balance downstream of melanocortin-4 receptor. Nat Neurosci 6:736–742PubMedCrossRef

31.

Unger TJ, Calderon GA, Bradley LC, Sena-Esteves M, Rios M (2007) Selective deletion of Bdnf in the ventromedial and dorsomedial hypothalamus of adult mice results in hyperphagic behavior and obesity. J Neurosci 27:14265–14274PubMedCrossRef

32.

El Gharbawy AH, Adler-Wailes DC, Mirch MC et al (2006) Serum brain-derived neurotrophic factor concentrations in lean and overweight children and adolescents. J Clin Endocrinol Metab 91:3548–3552PubMedCrossRef

33.

Krabbe KS, Nielsen AR, Krogh-Madsen R et al (2007) Brain-derived neurotrophic factor (BDNF) and type 2 diabetes. Diabetologia 50:431–438PubMedCrossRef

34.

Kernie SG, Liebl DJ, Parada LF (2000) BDNF regulates eating behavior and locomotor activity in mice. EMBO J 19:1290–1300PubMedCrossRef

35.

Zaltsman Y, Shachnai L, Yivgi-Ohana N et al (2010) MTCH2/MIMP is a major facilitator of tBID recruitment to mitochondria. Nat Cell Biol 12:553–562PubMedCrossRef

36.

Kulyté A, Rydén M, Mejhert N et al (2011) MTCH2 in human white adipose tissue and obesity. J Clin Endocrinol Metab 96:E1661–E1165PubMedCrossRef

37.

Gutierrez-Aguilar R, Kim DH, Woods SC, Seeley RJ (2012) Expression of new loci associated with obesity in diet-induced obese rats: from genetics to physiology. Obes (Silver Spring) 20:306–312CrossRef

38.

Hamm JK, el Jack AK, Pilch PF, Farmer SR (1999) Role of PPAR gamma in regulating adipocyte differentiation and insulin-responsive glucose uptake. Ann NY Acad Sci 892:134–145PubMedCrossRef

39.

Vidal-Puig AJ, Considine RV, Jimenez-Linan M et al (1997) Peroxisome proliferator-activated receptor gene expression in human tissues. Effects of obesity, weight loss, and regulation by insulin and glucocorticoids. J Clin Invest 99:2416–2422PubMedCrossRef

40.

Funatsu N, Miyata S, Kumanogoh H et al (1999) Characterization of a novel rat brain glycosylphosphatidylinositol-anchored protein (Kilon), a member of the IgLON cell adhesion molecule family. J Biol Chem 274:8224–8230PubMedCrossRef

41.

Hashimoto T, Yamada M, Maekawa S, Nakashima T, Miyata S (2008) IgLON cell adhesion molecule Kilon is a crucial modulator for synapse number in hippocampal neurons. Brain Res 1224:1–11PubMedCrossRef

42.

Walley AJ, Jacobson P, Falchi M et al (2012) Differential coexpression analysis of obesity-associated networks in human subcutaneous adipose tissue. Int J Obes (Lond) 36:137–147CrossRef

43.

Nishikawa K, Nakashima T, Takeda S et al (2010) Maf promotes osteoblast differentiation in mice by mediating the age-related switch in mesenchymal cell differentiation. J Clin Invest 120:3455–3465PubMedCrossRef

44.

Ren D, Li M, Duan C, Rui L (2005) Identification of SH2-B as a key regulator of leptin sensitivity, energy balance, and body weight in mice. Cell Metabol 2:95–104CrossRef

45.

Duan C, Yang H, White MF, Rui L (2004) Disruption of the SH2-B gene causes age-dependent insulin resistance and glucose intolerance. Mol Cell Biol 24:7435–7443PubMedCrossRef

46.

Ren D, Zhou Y, Morris D, Li M, Li Z, Rui L (2007) Neuronal SH2B1 is essential for controlling energy and glucose homeostasis. J Clin Invest 117:397–406PubMedCrossRef

47.

Garver WS, Francis GA, Jelinek D et al (2007) The National Niemann–Pick C1 disease database: report of clinical features and health problems. Am J Med Genet A 143A:1204–1211PubMedCrossRef

48.

Jelinek D, Millward V, Birdi A, Trouard TP, Heidenreich RA, Garver WS (2011) Npc1 haploinsufficiency promotes weight gain and metabolic features associated with insulin resistance. Hum Mol Genet 20:312–321PubMedCrossRef

49.

Veyrieras JB, Kudaravalli S, Kim SY et al (2008) High-resolution mapping of expression-QTLs yields insight into human gene regulation. PLoS Genet 4:e1000214PubMedCrossRef

50.

Park KW, Waki H, Choi SP, Park KM, Tontonoz P (2010) The small molecule phenamil is a modulator of adipocyte differentiation and PPARgamma expression. J Lipid Res 51:2775–2784PubMedCrossRef

Title: Functional relevance of genes implicated by obesity genome-wide association study signals for human adipocyte biology
Authors: F. Bernhard
K. Landgraf
N. Klöting
A. Berthold
P. Büttner
D. Friebe
W. Kiess
P. Kovacs
M. Blüher
A. Körner
Publication date: 01-02-2013
Publisher: Springer-Verlag
Published in: Diabetologia / Issue 2/2013
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-012-2773-0

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Functional relevance of genes implicated by obesity genome-wide association study signals for human adipocyte biology

Abstract

Aims/hypothesis

Methods

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Conclusions/interpretation

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Abstract

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Conclusions/interpretation

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