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Prediction of preadipocyte differentiation by gene expression reveals role of insulin receptor substrates and necdin

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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Figure 1: Considering phenotypes as a continuum.
Figure 2: Q-RT–PCR analyses confirm changes in gene expression in vitro and in vivo.
Figure 3: Role of necdin and E2Fs in brown adipogenesis.
Figure 4: Necdin RNAi restores differentiation, mitotic clonal expansion, and mRNA and protein expression.
Figure 5: Regulation of gene expression by CREB.
Figure 6: Proposed model of regulation of brown adipogenesis by the insulin/IGF-1/IRS pathway.

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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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Correspondence to C. Ronald Kahn.

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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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