ChREBP-β regulates thermogenesis in brown adipose tissue

in Journal of Endocrinology
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  • 1 Department of Pathophysiology, Naval Medical University, Shanghai, China
  • 2 Department of Physiology, Naval Medical University, Shanghai, China
  • 3 The State Key Laboratory of Membrane Biology, Center for Life Sciences and Institute of Molecular Medicine, Peking University, Beijing, China
  • 4 NHC Key Laboratory of Hormones and Development, Tianjin Key Laboratory of Metabolic Diseases, Tianjin Medical University Chu Hsien-I Memorial Hospital and Tianjin Institute of Endocrinology, Tianjin, China

Correspondence should be addressed to W J Zhang or W Wang: zbtb20@aliyun.com or wangwz68@163.com

*(C Wei, X Ma and K Su contributed equally to this work)

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Brown adipose tissue (BAT) plays a critical role in energy expenditure by uncoupling protein 1 (UCP1)-mediated thermogenesis. Carbohydrate response element-binding protein (ChREBP) is one of the key transcription factors regulating de novo lipogenesis (DNL). As a constitutively active form, ChREBP-β is expressed at extremely low levels. Up to date, its functional relevance in BAT remains unclear. In this study, we show that ChREBP-β inhibits BAT thermogenesis. BAT ChREBP-β mRNA levels were elevated upon cold exposure, which prompted us to generate a mouse model overexpressing ChREBP-β specifically in BAT using the Cre/LoxP approach. ChREBP-β overexpression led to a whitening phenotype of BAT at room temperature, as evidenced by increased lipid droplet size and decreased mitochondrion content. Moreover, BAT thermogenesis was inhibited upon acute cold exposure, and its metabolic remodeling induced by long-term cold adaptation was significantly impaired by ChREBP-β overexpression. Mechanistically, ChREBP-β overexpression downregulated expression of genes involved in mitochondrial biogenesis, autophagy, and respiration. Furthermore, thermogenic gene expression (e.g. Dio2, UCP1) was markedly inhibited in BAT by the overexpressed ChREBP-β. Put together, our work points to ChREBP-β as a negative regulator of thermogenesis in brown adipocytes.

Supplementary Materials

    • Supplementary Table 1. Information about the primary antibodies
    • Supplementary Table 2. Primers sequence for ChIP analysis
    • Supplementary Figure 1. Nuclear location of the overexpressed ChREBP- in BAT. Adult male BET- and control mice on normal chow were housed at room temperature or subjected to chronic cold training at 4°C for 2 weeks, then BAT was subjected to immunochemistry analysis. ChREBP was detected with an anti-ChREBP antibody and visualized by an Alex-594-conjugated secondary antibody in BAT from BET- and WT mice housed at room temperature (A) or subjected to cold training (B). The nuclei were counterstained with DAPI. The images were representative of three independent experiments. Scale bars, 50 m.
    • Supplementary Figure 2. Binding of ChREBP to target gene promoters in ChREBP- overexpressing BAT. BET- and control mice were housed at room temperature and fed normal chow diet. Their BAT was removed and subjected to ChIP analysis using anti-ChREBP antibodies or isotype IgG as control. NC1, negative control.
    • Supplementary Figure 3. ChREBP- overexpression does not affect energy and glucose homeostasis of mice on normal chow diet. Male (A-E) or female (F-J) BET- and WT control mice at the age of 10 weeks were analyzed when they were fed normal chow diet. Body weight (A, F), ratio of tissue weight to body weight (B, G), and blood glucose levels in fed (C, H) and 6 hour-fasting (D, J) state (n=5-6 per group). iWAT, inguinal WAT; eWAT, epididymal WAT. (E, J) Glucose tolerance test with 2g/kg glucose (n=6 per group). Data are presented as mean ± SEM. *, p<0.05; **, p<0.01.
    • Supplementary Figure 4. ChREBP- overexpression improves glucose tolerance in mice fed high fat diet. Male BET- and WT control mice were fed a high-fat diet (HFD, 60% fat in calorie) for 15 weeks starting from 4 weeks of age. (A) Growth curve of BET- and control mice (n=6 per group). (B) Glucose tolerance test with 2g/kg glucose (n=6 per group). Data are presented as mean ± SEM. *, p<0.05.
    • Supplementary Figure 5. The effects of ChREBP- overexpression on the mRNA levels of genes involved in lipid metabolism in BAT. Adult male BET- and WT mice on normal chow were housed at room temperature or subjected to 6-hour acute cold exposure or 2-week cold adaptation at 4°C (n=6 per group). mRNA expression levels in BAT were analyzed and normalized with the internal control -actin. Data are presented as mean ± SEM. *, p<0.05; **, p<0.01; ***, p<0.001.

 

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