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An ERK/Cdk5 axis controls the diabetogenic actions of PPARγ

Nature volume 517, pages 391–395 (2015)Cite this article

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Abstract

Obesity-linked insulin resistance is a major precursor to the development of type 2 diabetes. Previous work has shown that phosphorylation of PPARγ (peroxisome proliferator-activated receptor γ) at serine 273 by cyclin-dependent kinase 5 (Cdk5) stimulates diabetogenic gene expression in adipose tissues1. Inhibition of this modification is a key therapeutic mechanism for anti-diabetic drugs that bind PPARγ, such as the thiazolidinediones and PPARγ partial agonists or non-agonists2. For a better understanding of the importance of this obesity-linked PPARγ phosphorylation, we created mice that ablated Cdk5 specifically in adipose tissues. These mice have both a paradoxical increase in PPARγ phosphorylation at serine 273 and worsened insulin resistance. Unbiased proteomic studies show that extracellular signal-regulated kinase (ERK) kinases are activated in these knockout animals. Here we show that ERK directly phosphorylates serine 273 of PPARγ in a robust manner and that Cdk5 suppresses ERKs through direct action on a novel site in MAP kinase/ERK kinase (MEK). Importantly, pharmacological inhibition of MEK and ERK markedly improves insulin resistance in both obese wild-type and ob/ob mice, and also completely reverses the deleterious effects of the Cdk5 ablation. These data show that an ERK/Cdk5 axis controls PPARγ function and suggest that MEK/ERK inhibitors may hold promise for the treatment of type 2 diabetes.

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Figure 1: Insulin resistance after Cdk5 deletion in adipocytes.
Figure 2: Identification and characterization of ERK as a S273 PPARγ kinase.
Figure 3: Regulation of MEK2 by Cdk5.
Figure 4: Metabolic consequences of MEK inhibition in vivo in mice fed with a high-fat diet.
Figure 5: Metabolic consequences of MEK inhibition in ob/ob mice.

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Acknowledgements

We thank E. Rosen for providing us with the adiponectin Cre mice before their initial publication; members of the Spiegelman laboratory (Dana-Farber Cancer Institute) and D. Cohen (Brigham and Women's Hospital) for discussions; and C. Palmer and K. LeClair for reading the manuscript. B.M.S. acknowledges National Institutes of Health (NIH) grant DK31405. A.B. acknowledges NIH grant DK93638, the Harvard University Milton Fund, and the Harvard Digestive Disease Center, Core D.

Author information

Authors and Affiliations

  1. Division of Endocrinology, Diabetes and Hypertension, Brigham and Women's Hospital and Harvard Medical School, Boston, 02115, Massachusetts, USA

    Alexander S. Banks & Peter-James H. Zushin

  2. Department of Cell Biology, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Fiona E. McAllister, Steven P. Gygi & Bruce M. Spiegelman

  3. Yale Mouse Metabolic Phenotyping Center and Departments of Internal Medicine and Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, 06510, Connecticut, USA

    João Paulo G. Camporez, Michael J. Jurczak & Gerald I. Shulman

  4. Dana-Farber Cancer Institute, Boston, 02115, Massachusetts, USA

    Dina Laznik-Bogoslavski & Bruce M. Spiegelman

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  1. Alexander S. Banks
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Contributions

A.B., B.M.S., F.E., S.G., J.P.C., M.J. and G.S. designed the experiments. A.B., D.B., F.E., J.C.P. and P.Z. performed the experiments. A.B., B.M.S. and F.E. wrote the manuscript.

Corresponding authors

Correspondence to Alexander S. Banks or Bruce M. Spiegelman.

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Competing interests

B.M.S. is a consultant to and shareholder in Ember Therapeutics. The remaining authors declare no competing interests.

Extended data figures and tables

Extended Data Figure 1 Metabolic profiling of adipose-specific Cdk5-KO mice on a standard chow diet.

a–e, Fasting plasma levels of glucose (a), insulin (b), total triacylglycerols (c), free fatty acids (FFA) (d) and total cholesterol (e) (n = 16 (control) and 17 (KO)). f, g, Body weights (f) and intraperitoneal glucose tolerance test (g). Mice were 12 weeks of age (n = 14 (control) and 11 (KO)). No significant differences were observed. Error bars indicate s.e.m.

Extended Data Figure 2 Energy homeostasis of adipose-specific Cdk5-KO mice maintained on a high-fat diet.

a–f, After a 48-h acclimatization period, singly housed mice were monitored for oxygen consumption (VO2) (a), carbon dioxide production (VCO2) (b), respiratory exchange ratio (RER) (c), ambulatory locomotor activity (d), cumulative food intake (e) and body weights (f) (n = 8 per group). Shaded areas signify the dark phase of the light cycle. No significant differences were observed. Error bars indicate s.e.m.

Extended Data Figure 3 Activity of alternative kinases in adipose tissue from Cdk5-KO mice.

Brown adipose tissue protein lysates from mice maintained on a high-fat diet for 12 weeks. Blotting for phospho-p38, phospho-JNK and phospho-S473 and pT308 AKT was performed before loading for total protein amounts.

Extended Data Figure 4 Conservation of the sites on MEK2 phosphorylated by Cdk5.

Mouse MEK2 T395/T397 corresponds to human MEK2 T394/T396. These sites share identity with MEK1 T386/T388 in both humans and mouse. Cdk5 has previously been shown to phosphorylate MEK1 at T286, a site not shared with MEK2. ERK has been shown to phosphorylate MEK1 T386 and contribute to regulation of kinase activity31. Homo, Homo sapiens; trog, Pan troglodytes; mus, Mus musculus; rat, Rattus norvegicus; bos, Bos taurus; canis, Canis lupus familiaris.

Extended Data Figure 5 Body weight of control and of adipose-specific Cdk5-KO mice maintained on a high-fat diet after treatment with PD0325901.

Treatment similar to that in Fig. 4a–c. The body weights are not significantly different by ANOVA. Error bars indicate s.e.m.

Extended Data Figure 6 Effects of PD0325901 treatment on ob/ob mice.

a–c, Glucose tolerance test (a), adiponectin levels (b) and body weights (c) of ob/ob mice treated with PD0325901 (n = 7 (vehicle) and 8 (PD)). *P ≤ 0.05 by Student’s t-test. Error bars indicate s.e.m.

Extended Data Figure 7 Inflammatory markers in epididymal white adipose tissue from ob/ob mice treated with MEK inhibitors.

Gene expression analysis was performed on M1 macrophage markers Nos2 and tumour necrosis factor-α (TNF-α); M2 macrophage markers Arg1, Chil3, Il10, Itgax, Clec10a/Mgl1 and Mgl2; chemotactic ligand Ccl2 and receptor Ccr2; and macrophage surface markers Emr1, Cd68 and Csf1r (n = 7 or 8 mice per group as in Fig. 5f, h). Gene expression was analysed by ANOVA. Error bars indicate s.e.m. *P < 0.05; **P < 0.01; ***P < 0.001.

Extended Data Figure 8 Schematic model of PPARγ regulation at S273.

a, In the lean state, PPARγ is not phosphorylated. b, In the obese state, S273 phosphorylation is driven by both Cdk5 and ERK with CDK5 repressing MEK and ERK activity. c, Cdk5-KO results in derepression of MEK and ERK kinases and increased phosphorylation of S273 PPARγ. d, MEK inhibition markedly decreases S273 PPARγ phosphorylation. e, PPARγ ligands, including the thiazolidinediones, block the accessibility of S273 PPARγ by either ERK or CDK5 kinases.

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Banks, A., McAllister, F., Camporez, J. et al. An ERK/Cdk5 axis controls the diabetogenic actions of PPARγ. Nature 517, 391–395 (2015). https://doi.org/10.1038/nature13887

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