Abstract
The insulin/IGF-1 (insulin-like growth factor 1) signalling pathway promotes adipocyte differentiation via complex signalling networks. Here, using microarray analysis of brown preadipocytes that are derived from wild-type and insulin receptor substrate (Irs) knockout animals that exhibit progressively impaired differentiation, we define 374 genes/expressed-sequence tags whose expression in preadipocytes correlates with the ultimate ability of the cells to differentiate. Many of these genes, including preadipocyte factor-1 (Pref-1) and multiple members of the Wnt signalling pathway, are related to early adipogenic events. Necdin is also markedly increased in Irs knockout cells that cannot differentiate, and knockdown of necdin restores brown adipogenesis with downregulation of Pref-1 and Wnt10a expression. Insulin receptor substrate proteins regulate a necdin–E2F4 interaction that represses peroxisome-proliferator-activated receptor γ (PPARγ) transcription via a cyclic AMP response element binding protein (CREB)-dependent pathway. Together these define a key signalling network that is involved in brown preadipocyte determination.
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References
Rangwala, S. M. & Lazar, M. A. Transcriptional control of adipogenesis. Annu. Rev. Nutr. 20, 535–559 (2000).
Koutnikova, H. & Auwerx, J. Regulation of adipocyte differentiation. Ann. Med. 33, 556–561 (2001).
MacDougald, O. A. & Mandrup, S. Adipogenesis: forces that tip the scales. Trends Endocrinol. Metab. 13, 5–11 (2002).
Gregoire, F. M. Adipocyte differentiation: from fibroblast to endocrine cell. Exp. Biol. Med. (Maywood) 226, 997–1002 (2001).
Cowherd, R. M., Lyle, R. E. & McGehee, R. E. J. Molecular regulation of adipocyte differentiation. PMID 10, 3–10 (1999).
Rosen, E. D., Walkey, C. J., Puigserver, P. & Spiegelman, B. M. Transcriptional regulation of adipogenesis. Genes Dev. 14, 1293–1307 (2000).
Puigserver, P. et al. A cold-inducible coactivator of nuclear receptors linked to adaptive thermogenesis. Cell 92, 829–839 (1998).
Gregoire, F. M., Smas, C. M. & Sul, H. S. Understanding adipocyte differentiation. Physiol. Rev. 78, 783–809 (1998).
Ross, S. E. et al. Inhibition of adipogenesis by Wnt signaling. Science 289, 950–953 (2000).
Smas, C. M. & Sul, H. S. Pref-1, a protein containing EGF-like repeats, inhibits adipocyte differentiation. Cell 73, 725–734 (1993).
Tong, Q. et al. Function of GATA transcription factors in preadipocyte-adipocyte transition. Science 290, 134–138 (2000).
Chen, P. L., Riley, D. J., Chen, Y. & Lee, W. H. Retinoblastoma protein positively regulates terminal adipocyte differentiation through direct interaction with C/EBPs. Genes Dev. 10, 2794–2804 (1996).
Tseng, Y. H., Kriauciunas, K. M., Kokkotou, E. & Kahn, C. R. Differential roles of insulin receptor substrates in brown adipocyte differentiation. Mol. Cell. Biol. 24, 1918–1929 (2004).
Bartholomew, D. J. A test of homogeneity for ordered alternatives. Biometrika 46, 36–48 (1959).
Storey, J. D. & Tibshirani, R. Statistical significance for genomewide studies. Proc. Natl Acad. Sci. USA 100, 9440–9445 (2003).
Spiegelman, B. M. & Farmer, S. R. Decreases in tubulin and actin gene expression prior to morphological differentiation of 3T3 adipocytes. Cell 29, 53–60 (1982).
Weiner, F. R., Shah, A., Smith, P. J., Rubin, C. S. & Zern, M. A. Regulation of collagen gene expression in 3T3-L1 cells. Effects of adipocyte differentiation and tumor necrosis factor alpha. Biochemistry 28, 4094–4099 (1989).
Yi, T., Choi, H. M., Park, R. W., Sohn, K. Y. & Kim, I. S. Transcriptional repression of type I procollagen genes during adipocyte differentiation. Exp. Mol. Med. 33, 269–275 (2001).
Vu, T. H. & Werb, Z. Matrix metalloproteinases: effectors of development and normal physiology. Genes Dev. 14, 2123–2133 (2000).
Chavey, C. et al. Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation. J. Biol. Chem. 278, 11888–11896 (2003).
Maquoi, E., Munaut, C., Colige, A., Collen, D. & Lijnen, H. R. Modulation of adipose tissue expression of murine matrix metalloproteinases and their tissue inhibitors with obesity. Diabetes 51, 1093–1101 (2002).
Schiller, P. C., D'Ippolito, G., Brambilla, R., Roos, B. A. & Howard, G. A. Inhibition of gap-junctional communication induces the trans-differentiation of osteoblasts to an adipocytic phenotype in vitro. J. Biol. Chem. 276, 14133–14138 (2001).
Kang, S. et al. Effects of Wnt signaling on brown adipocyte differentiation and metabolism mediated by PGC-1α. Mol. Cell. Biol. 25, 1272–1282 (2005).
Kawano, Y. & Kypta, R. Secreted antagonists of the Wnt signalling pathway. J. Cell Sci. 116, 2627–2634 (2003).
Escalante-Alcalde, D. et al. The lipid phosphatase LPP3 regulates extra-embryonic vasculogenesis and axis patterning. Development 130, 4623–4637 (2003).
Mata, J., Curado, S., Ephrussi, A. & Rorth, P. Tribbles coordinates mitosis and morphogenesis in Drosophila by regulating string/CDC25 proteolysis. Cell 101, 511–522 (2000).
Du, K., Herzig, S., Kulkarni, R. N. & Montminy, M. TRB3: a tribbles homolog that inhibits Akt/PKB activation by insulin in liver. Science 300, 1574–1577 (2003).
Fasshauer, M. et al. Essential role of insulin receptor substrate 1 in differentiation of brown adipocytes. Mol. Cell. Biol. 21, 319–329 (2001).
Araki, E. et al. Alternative pathway of insulin signaling in mice with targeted disruption of the IRS-1 gene. Nature 372, 186–190 (1994).
Tamemoto, H. et al. Insulin resistance and growth retardation in mice lacking insulin receptor substrate-1. Nature 372, 182–186 (1994).
Laustsen, P. G. et al. Lipoatrophic diabetes in Irs1−/−/Irs3−/− double knockout mice. Genes Dev. 16, 3213–3222 (2002).
Goldstone, A. P. Prader-Willi syndrome: advances in genetics, pathophysiology and treatment. Trends Endocrinol. Metab. 15, 12–20 (2004).
Taniura, H., Taniguchi, N., Hara, M. & Yoshikawa, K. Necdin, a postmitotic neuron-specific growth suppressor, interacts with viral transforming proteins and cellular transcription factor E2F1. J. Biol. Chem. 273, 720–728 (1998).
Kobayashi, M., Taniura, H. & Yoshikawa, K. Ectopic expression of necdin induces differentiation of mouse neuroblastoma cells. J. Biol. Chem. 277, 42128–42135 (2002).
Fajas, L. et al. E2Fs regulate adipocyte differentiation. Dev. Cell 3, 39–49 (2002).
Tseng, Y. H., Ueki, K., Kriauciunas, K. M. & Kahn, C. R. Differential roles of insulin receptor substrates in the anti-apoptotic function of insulin-like growth factor-1 and insulin. J. Biol. Chem. 277, 31601–31611 (2002).
Reusch, J. E., Colton, L. A. & Klemm, D. J. CREB activation induces adipogenesis in 3T3-L1 cells. Mol. Cell. Biol. 20, 1008–1020 (2000).
Klemm, D. J. et al. Insulin-induced adipocyte differentiation. Activation of CREB rescues adipogenesis from the arrest caused by inhibition of prenylation. J. Biol. Chem. 276, 28430–28435 (2001).
Landsberg, R. L. et al. The role of E2F4 in adipogenesis is independent of its cell cycle regulatory activity. Proc. Natl Acad. Sci. USA 100, 2456–2461 (2003).
Martelli, F. & Livingston, D. M. Regulation of endogenous E2F1 stability by the retinoblastoma family proteins. Proc. Natl Acad. Sci. USA 96, 2858–2863 (1999).
Yang, T. et al. A mouse model for Prader-Willi syndrome imprinting-centre mutations. Nature Genet. 19, 25–31 (1998).
Boeuf, S. et al. Differential gene expression in white and brown preadipocytes. Physiol. Genomics 7, 15–25 (2001).
Brunelli, S. et al. Msx2 and necdin combined activities are required for smooth muscle differentiation in mesoangioblast stem cells. Circ. Res. 94, 1571–1578 (2004).
Kuwajima, T., Taniura, H., Nishimura, I. & Yoshikawa, K. Necdin interacts with the Msx2 homeodomain protein via MAGE-D1 to promote myogenic differentiation of C2C12 cells. J. Biol. Chem. 279, 40484–40493 (2004).
Jansson, P. A. et al. A novel cellular marker of insulin resistance and early atherosclerosis in humans is related to impaired fat cell differentiation and low adiponectin. FASEB J. 17, 1434–1440 (2003).
Asahara, H. et al. Chromatin-dependent cooperativity between constitutive and inducible activation domains in CREB. Mol. Cell. Biol. 21, 7892–7900 (2001).
Press, W. H., Teukolsky, S. A., Vetterling, W. T. & Flannery, B. P. Numerical Recipes in C: The Art of Scientific Computing (Cambridge Univ. Press, Cambridge, 1993).
Pan, W., Lin, J. & Le, C. T. How many replicates of arrays are required to detect gene expression changes in microarray experiments? A mixture model approach. Genome Biol. 3, research0022 (2002).
Ideker, T., Thorsson, V., Siegel, A. F. & Hood, L. E. Testing for differentially-expressed genes by maximum-likelihood analysis of microarray data. J. Comput. Biol. 7, 805–817 (2000).
Levene, H. Contributions to Probability and Statistics: Essays in Honor of Harold Hotelling (eds Olkin, I. et al.) 278–292 (Stanford Univ. Press, Stanford, CA, 1960).
Acknowledgements
We acknowledge A. Norris for constructive comments on the manuscript. We thank M. Montminy (Salk Institute for Biological Studies, La Jolla, CA) and L. Fajas (Metabolism and Cancer Laboratory, INSERM, France) for providing the plasmids used in this study. We acknowledge J. Klein and M. Fasshauer for preparation of cell lines, P. Laustsen, S. Crunkhorn, B. Emanuelli, S. Gesta, D. Espinoza, P. Lin and H. Gami for technical assistance, and J. Marr for excellent secretarial assistance. This work was supported in part by the National Institutes of Health grants DK33201, DK60837 (to C.R.K.), DK101183 (to Y.-H.T.), DK63696 (to A.J.B.) and DK07260 (to A.M.C.) as well as grants from the Lawson Wilkins Pediatric Endocrine Society and the Harvard/MIT Health Sciences and Technology (to A.J.B.).
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Tseng, YH., Butte, A., Kokkotou, E. et al. Prediction of preadipocyte differentiation by gene expression reveals role of insulin receptor substrates and necdin. Nat Cell Biol 7, 601–611 (2005). https://doi.org/10.1038/ncb1259
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DOI: https://doi.org/10.1038/ncb1259
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