Obesity is associated with macrophage accumulation in adipose tissue

Stuart Weisberg

Daniel McCann

Manisha Desai

Michael Rosenbaum

Rudolph Leibel

Anthony Ferrante

^{Division of Molecular Genetics, Department of Pediatrics,}^{Department of Biostatistics,}^{Deparment of Medicine, and}^{Naomi Berrie Diabetes Center, Columbia University, New York, New York, USA}

Address correspondence to: Anthony W. Ferrante, Jr., Naomi Berrie Diabetes Center, Columbia University, 1150 St. Nicholas Avenue, New York, New York 10032, USA. Phone: (212) 851-5322; Fax: (212) 851-5331; E-mail: ude.aibmuloc@7fwa.

Received 2003 Jun 19; Accepted 2003 Oct 13.

Abstract

Obesity alters adipose tissue metabolic and endocrine function and leads to an increased release of fatty acids, hormones, and proinflammatory molecules that contribute to obesity associated complications. To further characterize the changes that occur in adipose tissue with increasing adiposity, we profiled transcript expression in perigonadal adipose tissue from groups of mice in which adiposity varied due to sex, diet, and the obesity-related mutations agouti (Ay) and obese (Lepob). We found that the expression of 1,304 transcripts correlated significantly with body mass. Of the 100 most significantly correlated genes, 30% encoded proteins that are characteristic of macrophages and are positively correlated with body mass. Immunohistochemical analysis of perigonadal, perirenal, mesenteric, and subcutaneous adipose tissue revealed that the percentage of cells expressing the macrophage marker F4/80 (F4/80^{) was significantly and positively correlated with both adipocyte size and body mass. Similar relationships were found in human subcutaneous adipose tissue stained for the macrophage antigen CD68. Bone marrow transplant studies and quantitation of macrophage number in adipose tissue from macrophage-deficient (}Csf1op/op) mice suggest that these F4/80^{cells are CSF-1 dependent, bone marrow–derived adipose tissue macrophages. Expression analysis of macrophage and nonmacrophage cell populations isolated from adipose tissue demonstrates that adipose tissue macrophages are responsible for almost all adipose tissue TNF-α expression and significant amounts of iNOS and IL-6 expression. Adipose tissue macrophage numbers increase in obesity and participate in inflammatory pathways that are activated in adipose tissues of obese individuals.}

Abstract

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Acknowledgments

We thank E. Richard Stanley for the gift of FVB/NJ Csf1op/op mice. This work was supported by grants from the NIH NIDDK (K08 DK-59960 and R01 DK-66525 to A.W. Ferrante, Jr.; R01 DK-052431 to R.L. Leibel), the Fawcett Foundation, and the Russell Berrie Foundation. We thank Domenico Accili, E. Richard Stanley, and Yiying Zhang for their comments and suggestions.

Acknowledgments

Footnotes

See the related Commentary beginning on page 1785.

Conflict of interest: The authors have declared that no conflict of interest exists.

Nonstandard abbreviations used: monocyte chemotactic protein-1 (MCP-1); plasminogen activator inhibitor type 1 (PAI-1); stromal vascular cell (SVC); diet-induced obese (DIO); fatty acid–poor BSA (FAP-BSA); colony-stimulating factor 1 receptor (Csf1r); thiazolidinedione (TZD).

Footnotes

References

1. Tai ES, Lau TN, Ho SC, Fok AC, Tan CE. Body fat distribution and cardiovascular risk in normal weight women. Associations with insulin resistance, lipids and plasma leptin. Int. J. Obes. Relat. Metab. Disord.2000;24:751–757.[PubMed]
2. Messerli FH, et al. Obesity and essential hypertension. Hemodynamics, intravascular volume, sodium excretion, and plasma renin activity. Arch. Intern. Med.1981;141:81–85.[PubMed]
3. Liuzzi A, et al Serum leptin concentration in moderate and severe obesity: relationship with clinical, anthropometric and metabolic factors. Int. J. Obes. Relat. Metab. Disord.1999;23:1066–1073.[PubMed]
4. Janssen I, Katzmarzyk PT, Ross RBody mass index, waist circumference, and health risk: evidence in support of current National Institutes of Health guidelines. Arch. Intern. Med.2002;162:2074–2079.[PubMed]
5. Stolk RP, Meijer R, Mali WP, Grobbee DE, van der Graaf YUltrasound measurements of intraabdominal fat estimate the metabolic syndrome better than do measurements of waist circumference. Am. J. Clin. Nutr.2003;77:857–860.[PubMed]
6. DiPietro L, Katz LD, Nadel ERExcess abdominal adiposity remains correlated with altered lipid concentrations in healthy older women. Int. J. Obes. Relat. Metab. Disord.1999;23:432–436.[PubMed]
7. Hubert HB, Feinleib M, McNamara PM, Castelli WPObesity as an independent risk factor for cardiovascular disease: a 26-year follow-up of participants in the Framingham Heart Study. Circulation.1983;67:968–977.[PubMed]
8. Kurth T, et al Body mass index and the risk of stroke in men. Arch. Intern. Med.2002;162:2557–2562.[PubMed]
9. Calle EE, Rodriguez C, Walker-Thurmond K, Thun MJ. Overweight, obesity, and mortality from cancer in a prospectively studied cohort of U.S. adults. N. Engl. J. Med.2003;348:1625–1638.[PubMed]
10. Rosenbaum M, Leibel RLThe role of leptin in human physiology. N. Engl. J. Med.1999;341:913–915.[PubMed]
11. Maffei M, et al Increased expression in adipocytes of ob RNA in mice with lesions of the hypothalamus and with mutations at the db locus. Proc. Natl. Acad. Sci. U. S. A.1995;92:6957–6960.
12. Frederich RC, et al. Expression of ob mRNA and its encoded protein in rodents. Impact of nutrition and obesity. J. Clin. Invest.1995;96:1658–1663.
13. Zhang Y, Guo KY, Diaz PA, Heo M, Leibel RLDeterminants of leptin gene expression in fat depots of lean mice. Am. J. Physiol. Regul. Integr. Comp. Physiol.2002;282:R226–R234.[PubMed]
14. Bodary PF, Westrick RJ, Wickenheiser KJ, Shen Y, Eitzman DTEffect of leptin on arterial thrombosis following vascular injury in mice. JAMA.2002;287:1706–1709.[PubMed]
15. Arita Y, et al Paradoxical decrease of an adipose-specific protein, adiponectin, in obesity. Biochem. Biophys. Res. Commun.1999;257:79–83.[PubMed]
16. Combs TP, Berg AH, Obici S, Scherer PE, Rossetti LEndogenous glucose production is inhibited by the adipose-derived protein Acrp30. J. Clin. Invest.2001;108:1875–1881. doi:10.1172/JCI200114120.
17. Tomas E, et al Enhanced muscle fat oxidation and glucose transport by ACRP30 globular domain: acetyl-CoA carboxylase inhibition and AMP-activated protein kinase activation. Proc. Natl. Acad. Sci. U. S. A.2002;99:16309–16313.
18. Hotamisligil GS, Shargill NS, Spiegelman BMAdipose expression of tumor necrosis factor-alpha: direct role in obesity-linked insulin resistance. Science.1993;259:87–91.[PubMed]
19. Fried SK, Bunkin DA, Greenberg ASOmental and subcutaneous adipose tissues of obese subjects release interleukin-6: depot difference and regulation by glucocorticoid. J. Clin. Endocrinol. Metab.1998;83:847–850.[PubMed]
20. Vgontzas AN, et al Elevation of plasma cytokines in disorders of excessive daytime sleepiness: role of sleep disturbance and obesity. J. Clin. Endocrinol. Metab.1997;82:1313–1316.[PubMed]
21. Perreault M, Marette ATargeted disruption of inducible nitric oxide synthase protects against obesity-linked insulin resistance in muscle. Nat. Med.2001;7:1138–1143.[PubMed]
22. Samad F, Yamamoto K, Pandey M, Loskutoff DJElevated expression of transforming growth factor-beta in adipose tissue from obese mice. Mol. Med.1997;3:37–48.
23. Visser M, Bouter LM, McQuillan GM, Wener MH, Harris TBElevated C-reactive protein levels in overweight and obese adults. JAMA.1999;282:2131–2135.[PubMed]
24. Weyer C, et al Humoral markers of inflammation and endothelial dysfunction in relation to adiposity and in vivo insulin action in Pima Indians. Atherosclerosis.2002;161:233–242.[PubMed]
25. Sartipy P, Loskutoff DJMonocyte chemoattractant protein 1 in obesity and insulin resistance. Proc. Natl. Acad. Sci. U. S. A.2003;100:7265–7270.
26. Samad F, Yamamoto K, Loskutoff DJ. Distribution and regulation of plasminogen activator inhibitor-1 in murine adipose tissue in vivo. Induction by tumor necrosis factor-alpha and lipopolysaccharide. J. Clin. Invest.1996;97:37–46.
27. Samad F, Pandey M, Loskutoff DJTissue factor gene expression in the adipose tissues of obese mice. Proc. Natl. Acad. Sci. U. S. A.1998;95:7591–7596.
28. De Pergola G, Pannacciulli NCoagulation and fibrinolysis abnormalities in obesity. J. Endocrinol. Invest.2002;25:899–904.[PubMed]
29. Uysal KT, Wiesbrock SM, Marino MW, Hotamisligil GSProtection from obesity-induced insulin resistance in mice lacking TNF-alpha function. Nature.1997;389:610–614.[PubMed]
30. Hotamisligil GS, Murray DL, Choy LN, Spiegelman BMTumor necrosis factor alpha inhibits signaling from the insulin receptor. Proc. Natl. Acad. Sci. U. S. A.1994;91:4854–4858.
31. Zhang HH, Halbleib M, Ahmad F, Manganiello VC, Greenberg ASTumor necrosis factor-alpha stimulates lipolysis in differentiated human adipocytes through activation of extracellular signal-related kinase and elevation of intracellular cAMP. Diabetes.2002;51:2929–2935.[PubMed]
32. Nonogaki K, et al Interleukin-6 stimulates hepatic triglyceride secretion in rats. Endocrinology.1995;136:2143–2149.[PubMed]
33. Ross SE, et al Microarray analyses during adipogenesis: understanding the effects of Wnt signaling on adipogenesis and the roles of liver X receptor alpha in adipocyte metabolism. Mol. Cell. Biol.2002;22:5989–5999.
34. Fain JN, Cheema PS, Bahouth SW, Lloyd Hiler MResistin release by human adipose tissue explants in primary culture. Biochem. Biophys. Res. Commun.2003;300:674–678.[PubMed]
35. Bultman SJ, Michaud EJ, Woychik RPMolecular characterization of the mouse agouti locus. Cell.1992;71:1195–1204.[PubMed]
36. Leibel RL, Chung WK, Chua SC., Jr The molecular genetics of rodent single gene obesities. J. Biol. Chem.1997;272:31937–31940.[PubMed]
37. Robinson SW, Dinulescu DM, Cone RDGenetic models of obesity and energy balance in the mouse. Annu. Rev. Genet.2000;34:687–745.[PubMed]
38. Rosenbaum M, et al Effects of experimental weight perturbation on skeletal muscle work efficiency in human subjects. Am. J. Physiol. Regul. Integr. Comp. Physiol.2003;285:R183–R192.[PubMed]
39. Rosenbaum M, Murphy EM, Heymsfield SB, Matthews DE, Leibel RLLow dose leptin administration reverses effects of sustained weight-reduction on energy expenditure and circulating concentrations of thyroid hormones. J. Clin. Endocrinol. Metab.2002;87:2391–2394.[PubMed]
40. Leibel RL, Rosenbaum M, Hirsch JChanges in energy expenditure resulting from altered body weight. N. Engl. J. Med.1995;332:621–628.[PubMed]
41. Benjamini Y, Hochberg YControlling the false discovery rate: a practical and powerful approach to multiple testing. Journal of the Royal Statistical Society, Series B.1995;57:289–300.[PubMed]
42. Slonim DKFrom patterns to pathways: gene expression data analysis comes of age. Nat. Genet.2002;32(Suppl.):502–508.[PubMed]
43. Cecchini MG, et al Role of colony stimulating factor-1 in the establishment and regulation of tissue macrophages during postnatal development of the mouse. Development.1994;120:1357–1372.[PubMed]
44. Vandesompele J, et al Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol.2002;3:RESEARCH0034.
45. Rosen, S., and Skaletsky, H. 2000. Primer3 on the WWW for general users and for biologist programmers. In Bioinformatics methods and protocols: methods in molecular biology. S. Misener and S.A. Krawetz, editors. Humana Press. Totowa, New Jersey, USA. 365–386. [[PubMed]
46. Snedecor, G.W., and Cochran, W.G1967. Statistical methods. Iowa State University Press. Ames, Iowa, USA. 172–198.
47. Prieditis H, Adamson IYAlveolar macrophage kinetics and multinucleated giant cell formation after lung injury. J. Leukoc. Biol.1996;59:534–538.[PubMed]
48. Toyosaki-Maeda T, et al Differentiation of monocytes into multinucleated giant bone-resorbing cells: two-step differentiation induced by nurse-like cells and cytokines. Arthritis Res.2001;3:306–310.
49. Athanasou NA, Quinn JImmunophenotypic differences between osteoclasts and macrophage polykaryons: immunohistological distinction and implications for osteoclast ontogeny and function. J. Clin. Pathol.1990;43:997–1003.
50. Endemann G, et al CD36 is a receptor for oxidized low density lipoprotein. J. Biol. Chem.1993;268:11811–11816.[PubMed]
51. Lee CH, Evans RMPeroxisome proliferator-activated receptor-gamma in macrophage lipid homeostasis. Trends Endocrinol. Metab.2002;13:331–335.[PubMed]
52. Makowski L, et al Lack of macrophage fatty-acid-binding protein aP2 protects mice deficient in apolipoprotein E against atherosclerosis. Nat. Med.2001;7:699–705.
53. Cousin B, et al A role for preadipocytes as macrophage-like cells. FASEB J.1999;13:305–312.[PubMed]
54. Wiktor-Jedrzejczak W, et al Total absence of colony-stimulating factor 1 in the macrophage-deficient osteopetrotic (op/op) mouse. Proc. Natl. Acad. Sci. U. S. A.1990;87:4828–4832.
55. Yoshida H, et al The murine mutation osteopetrosis is in the coding region of the macrophage colony stimulating factor gene. Nature.1990;345:442–444.[PubMed]
56. Stanley ER, et al Biology and action of colony-stimulating factor-1. Mol. Reprod. Dev.1997;46:4–10.[PubMed]
57. Dai, X.M., Zong, X.H., Sylvestre, V., and Stanley, E.R. 2003. Incomplete restoration of colony stimulating factor-1 (CSF-1) function in CSF-1–deficient Csf1op/Csf1op mice by transgenic expression of cell surface CSF-1. Blood. doi:10.1182/blood-2003-08-2739. [[PubMed]
58. Kullo IJ, Hensrud DD, Allison TGComparison of numbers of circulating blood monocytes in men grouped by body mass index (<25, 25 to <30, > or = 30) Am. J. Cardiol.2002;89:1441–1443.[PubMed]
59. Soukas A, Cohen P, Socci ND, Friedman JMLeptin-specific patterns of gene expression in white adipose tissue. Genes Dev.2000;14:963–980.
60. Cousin B, Andre M, Casteilla L, Penicaud LAltered macrophage-like functions of preadipocytes in inflammation and genetic obesity. J. Cell. Physiol.2001;186:380–386.[PubMed]
61. Gordon SThe macrophage. Bioessays.1995;17:977–986.[PubMed]
62. Bornstein SR, et al Immunohistochemical and ultrastructural localization of leptin and leptin receptor in human white adipose tissue and differentiating human adipose cells in primary culture. Diabetes.2000;49:532–538.[PubMed]
63. Charriere G, et al. Preadipocyte conversion to macrophage. Evidence of plasticity. J. Biol. Chem.2003;278:9850–9855.[PubMed]
64. Gordon SThe role of the macrophage in immune regulation. Res. Immunol.1998;149:685–688.[PubMed]
65. Torres PF, et al Changes in cytokine mRNA levels in experimental corneal allografts after local clodronate-liposome treatment. Invest. Ophthalmol. Vis. Sci.1999;40:3194–3201.[PubMed]
66. Koay MA, et al Macrophages are necessary for maximal nuclear factor-kappa B activation in response to endotoxin. Am. J. Respir. Cell Mol. Biol.2002;26:572–578.[PubMed]
67. Pandey M, Tuncman G, Hotamisligil GS, Samad FDivergent roles for p55 and p75 TNF-alpha receptors in the induction of plasminogen activator inhibitor-1. Am. J. Pathol.2003;162:933–941.
68. Morrone G, et al Recombinant interleukin 6 regulates the transcriptional activation of a set of human acute phase genes. J. Biol. Chem.1988;263:12554–12558.[PubMed]
69. Pickup JC, Crook MAIs type II diabetes mellitus a disease of the innate immune system? Diabetologia.1998;41:1241–1248.[PubMed]
70. Rissanen P, Vahtera E, Krusius T, Uusitupa M, Rissanen AWeight change and blood coagulability and fibrinolysis in healthy obese women. Int. J. Obes. Relat. Metab. Disord.2001;25:212–218.[PubMed]
71. Saghizadeh M, Ong JM, Garvey WT, Henry RR, Kern PA. The expression of TNF alpha by human muscle. Relationship to insulin resistance. J. Clin. Invest.1996;97:1111–1116.
72. Kapur S, Bedar S, Marcotte B, Cote CH, Marette AExpression of nitric oxide synthase in skeletal muscle: a novel role for nitric oxide as a modulator of insulin action. Diabetes.1997;46:1691–1700.[PubMed]
73. Youd JM, Rattigan S, Clark MGAcute impairment of insulin-mediated capillary recruitment and glucose uptake in rat skeletal muscle in vivo by TNF-alpha. Diabetes.2000;49:1904–1909.[PubMed]
74. Schneider BS, Faust IM, Hemmes R, Hirsch JEffects of altered adipose tissue morphology on plasma insulin levels in the rat. Am. J. Physiol.1981;240:E358–E362.[PubMed]
75. Weyer C, et al Subcutaneous abdominal adipocyte size, a predictor of type 2 diabetes, is linked to chromosome 1q21—q23 and is associated with a common polymorphism in LMNA in Pima Indians. Mol. Genet. Metab.2001;72:231–238.[PubMed]
76. Weyer C, Foley JE, Bogardus C, Tataranni PA, Pratley REEnlarged subcutaneous abdominal adipocyte size, but not obesity itself, predicts type II diabetes independent of insulin resistance. Diabetologia.2000;43:1498–1506.[PubMed]
77. Stern JS, Batchelor BR, Hollander N, Cohn CK, Hirsch JAdipose-cell size and immunoreactive insulin levels in obese and normal-weight adults. Lancet.1972;2:948–951.[PubMed]
78. Jimenez J, Zuniga-Guajardo S, Zinman B, Angel AEffects of weight loss in massive obesity on insulin and C-peptide dynamics: sequential changes in insulin production, clearance, and sensitivity. J. Clin. Endocrinol. Metab.1987;64:661–668.[PubMed]
79. Kral JG, Bjorntorp P, Schersten T, Sjostrom LBody composition and adipose tissue cellularity before and after jejuno-ileostomy in severely obese subjects. Eur. J. Clin. Invest.1977;7:413–419.[PubMed]
80. Salans LB, Dougherty JW. The effect of insulin upon glucose metabolism by adipose cells of different size. Influence of cell lipid and protein content, age, and nutritional state. J. Clin. Invest.1971;50:1399–1410.
81. Hissin PJ, et al Mechanism of insulin-resistant glucose transport activity in the enlarged adipose cell of the aged, obese rat. J. Clin. Invest.1982;70:780–790.
82. Le Lay S, et al Cholesterol, a cell size-dependent signal that regulates glucose metabolism and gene expression in adipocytes. J. Biol. Chem.2001;276:16904–16910.[PubMed]
83. Oakes ND, Thalen PG, Jacinto SM, Ljung BThiazolidinediones increase plasma-adipose tissue FFA exchange capacity and enhance insulin-mediated control of systemic FFA availability. Diabetes.2001;50:1158–1165.[PubMed]
84. Maeda N, et al PPARgamma ligands increase expression and plasma concentrations of adiponectin, an adipose-derived protein. Diabetes.2001;50:2094–2099.[PubMed]
85. Tordjman J, et al Thiazolidinediones block fatty acid release by inducing glyceroneogenesis in fat cells. J. Biol. Chem.2003;278:18785–18790.[PubMed]
86. Guan HP, et al A futile metabolic cycle activated in adipocytes by antidiabetic agents. Nat. Med.2002;8:1122–1128.[PubMed]
87. Jiang C, Ting AT, Seed BPPAR-gamma agonists inhibit production of monocyte inflammatory cytokines. Nature.1998;391:82–86.[PubMed]
88. Chawla A, et al PPAR-gamma dependent and independent effects on macrophage-gene expression in lipid metabolism and inflammation. Nat. Med.2001;7:48–52.[PubMed]
89. Alleva DG, et al Regulation of murine macrophage proinflammatory and anti-inflammatory cytokines by ligands for peroxisome proliferator-activated receptor-gamma: counter-regulatory activity by IFN-gamma. J. Leukoc. Biol.2002;71:677–685.[PubMed]
90. Ricote M, Li AC, Willson TM, Kelly CJ, Glass CKThe peroxisome proliferator-activated receptor-gamma is a negative regulator of macrophage activation. Nature.1998;391:79–82.[PubMed]
91. Ross SR, Graves RA, Spiegelman BMTargeted expression of a toxin gene to adipose tissue: transgenic mice resistant to obesity. Genes Dev.1993;7:1318–1324.[PubMed]
92. Boring L, et al Impaired monocyte migration and reduced type 1 (Th1) cytokine responses in C-C chemokine receptor 2 knockout mice. J. Clin. Invest.1997;100:2552–2561.
93. Lu B, et al Abnormalities in monocyte recruitment and cytokine expression in monocyte chemoattractant protein 1-deficient mice. J. Exp. Med.1998;187:601–608.
94. Levine JA, Jensen MD, Eberhardt NL, O’Brien TAdipocyte macrophage colony-stimulating factor is a mediator of adipose tissue growth. J. Clin. Invest.1998;101:1557–1564.