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Cross-talk between hypoxia and insulin signaling through Phd3 regulates hepatic glucose and lipid metabolism and ameliorates diabetes

Nature Medicine volume 19, pages 1325–1330 (2013)Cite this article

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Abstract

Signaling initiated by hypoxia and insulin powerfully alters cellular metabolism. The protein stability of hypoxia-inducible factor-1 alpha (Hif-1α) and Hif-2α is regulated by three prolyl hydroxylase domain–containing protein isoforms (Phd1, Phd2 and Phd3). Insulin receptor substrate-2 (Irs2) is a critical mediator of the anabolic effects of insulin, and its decreased expression contributes to the pathophysiology of insulin resistance and diabetes1. Although Hif regulates many metabolic pathways2, it is unknown whether the Phd proteins regulate glucose and lipid metabolism in the liver. Here, we show that acute deletion of hepatic Phd3, also known as Egln3, improves insulin sensitivity and ameliorates diabetes by specifically stabilizing Hif-2α, which then increases Irs2 transcription and insulin-stimulated Akt activation. Hif-2α and Irs2 are both necessary for the improved insulin sensitivity, as knockdown of either molecule abrogates the beneficial effects of Phd3 knockout on glucose tolerance and insulin-stimulated Akt phosphorylation. Augmenting levels of Hif-2α through various combinations of Phd gene knockouts did not further improve hepatic metabolism and only added toxicity. Thus, isoform-specific inhibition of Phd3 could be exploited to treat type 2 diabetes without the toxicity that could occur with chronic inhibition of multiple Phd isoforms.

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Figure 1: Phd3 specifically regulates hepatic Hif-2α expression and glucose metabolism in vivo.
Figure 2: Worsened hepatotoxicity without improved metabolism in combination Phd knockout animals.
Figure 3: A Hif-2α –mediated increase in Irs2 expression in mice lacking hepatic Phd3 improves insulin action and reverses diabetes.
Figure 4: Both Hif-2 and Irs2 are required for improved metabolism in mice with a liver-specific knockout of Phd3.

References

  1. Taniguchi, C.M., Emanuelli, B. & Kahn, C.R. Critical nodes in signalling pathways: insights into insulin action. Nat. Rev. Mol. Cell Biol. 7, 85–96 (2006).

    Article  CAS  Google Scholar 

  2. Denko, N.C. Hypoxia, HIF1 and glucose metabolism in the solid tumour. Nat. Rev. Cancer 8, 705–713 (2008).

    Article  CAS  Google Scholar 

  3. Ivan, M. et al. HIF-α targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science 292, 464–468 (2001).

    Article  CAS  Google Scholar 

  4. Min, J.H. et al. Structure of an HIF-1α–pVHL complex: hydroxyproline recognition in signaling. Science 296, 1886–1889 (2002).

    Article  CAS  Google Scholar 

  5. Appelhoff, R.J. et al. Differential function of the prolyl hydroxylases PHD1, PHD2, and PHD3 in the regulation of hypoxia-inducible factor. J. Biol. Chem. 279, 38458–38465 (2004).

    Article  CAS  Google Scholar 

  6. Rankin, E.B. et al. The HIF signaling pathway in osteoblasts directly modulates erythropoiesis through the production of EPO. Cell 149, 63–74 (2012).

    Article  CAS  Google Scholar 

  7. Taniguchi, C.M., Ueki, K. & Kahn, R. Complementary roles of IRS-1 and IRS-2 in the hepatic regulation of metabolism. J. Clin. Invest. 115, 718–727 (2005).

    Article  CAS  Google Scholar 

  8. Minamishima, Y.A. et al. A feedback loop involving the Phd3 prolyl hydroxylase tunes the mammalian hypoxic response in vivo. Mol. Cell Biol. 29, 5729–5741 (2009).

    Article  CAS  Google Scholar 

  9. Horton, J.D., Goldstein, J.L. & Brown, M.S. SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver. J. Clin. Invest. 109, 1125–1131 (2002).

    Article  CAS  Google Scholar 

  10. Chen, C., Pore, N., Behrooz, A., Ismail-Beigi, F. & Maity, A. Regulation of glut1 mRNA by hypoxia-inducible factor-1. Interaction between H-ras and hypoxia. J. Biol. Chem. 276, 9519–9525 (2001).

    Article  CAS  Google Scholar 

  11. Minamishima, Y.A. et al. A feedback loop involving the Phd3 prolyl hydroxylase tunes the mammalian hypoxic response in vivo. Mol. Cell Biol. 29, 5729–5741 (2009).

    Article  CAS  Google Scholar 

  12. Kim, W.Y. et al. Failure to prolyl hydroxylate hypoxia-inducible factor α phenocopies VHL inactivation in vivo. EMBO J. 25, 4650–4662 (2006).

    Article  CAS  Google Scholar 

  13. Puigserver, P. et al. Insulin-regulated hepatic gluconeogenesis through FOXO1-PGC-1α interaction. Nature 423, 550–555 (2003).

    Article  CAS  Google Scholar 

  14. Vassen, L., Wegrzyn, W. & Klein-Hitpass, L. Human insulin receptor substrate-2: gene organization and promoter characterization. Diabetes 48, 1877–1880 (1999).

    Article  CAS  Google Scholar 

  15. Kim, W.Y. et al. Failure to prolyl hydroxylate hypoxia-inducible factor α phenocopies VHL inactivation in vivo. EMBO J. 25, 4650–4662 (2006).

    Article  CAS  Google Scholar 

  16. Nakagawa, Y. et al. TFE3 transcriptionally activates hepatic IRS-2, participates in insulin signaling and ameliorates diabetes. Nat. Med. 12, 107–113 (2006).

    Article  CAS  Google Scholar 

  17. Saito, T. et al. Transcriptional regulation of endochondral ossification by HIF-2α during skeletal growth and osteoarthritis development. Nat. Med. 16, 678–686 (2010).

    Article  CAS  Google Scholar 

  18. Kubota, N et al. Dynamic functional relay between insulin receptor substrate 1 and 2 in hepatic insulin signaling during fasting and feeding. Cell Metab. 8, 49–64 (2008).

    Article  CAS  Google Scholar 

  19. Canettieri, G. et al. Dual role of the coactivator TORC2 in modulating hepatic glucose output and insulin signaling. Cell Metab. 2, 331–338 (2005).

    Article  CAS  Google Scholar 

  20. Takeda, K. et al. Regulation of adult erythropoiesis by prolyl hydroxylase domain proteins. Blood 111, 3229–3235 (2008).

    Article  CAS  Google Scholar 

  21. Minamishima, Y.A. & Kaelin, W.G. Jr. Reactivation of hepatic EPO synthesis in mice after PHD loss. Science 329, 407 (2010).

    Article  CAS  Google Scholar 

  22. Shen, X. et al. Prolyl hydroxylase inhibitors increase neoangiogenesis and callus formation following femur fracture in mice. J. Orthop. Res. 27, 1298–1305 (2009).

    Article  CAS  Google Scholar 

  23. Myllyharju, J. Prolyl 4-hydroxylases, key enzymes in the synthesis of collagens and regulation of the response to hypoxia, and their roles as treatment targets. Ann. Med. 40, 402–417 (2008).

    Article  CAS  Google Scholar 

  24. Luo, J. et al. A protocol for rapid generation of recombinant adenoviruses using the AdEasy system. Nat. Protoc. 2, 1236–1247 (2007).

    Article  CAS  Google Scholar 

  25. Wei, K. et al. A liver Hif-2α–Irs2 pathway sensitizes hepatic insulin signaling and is modulated by Vegf inhibition. Nat. Med. 10.1038/nm.3295 (2013).

  26. Rankin, E.B. et al. Inactivation of the arylhydrocarbon receptor nuclear translocator (Arnt) suppresses von Hippel-Lindau disease–associated vascular tumors in mice. Mol. Cell Biol. 25, 3163–3172 (2005).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank K. Takeda and G.-H. Fong (University of Connecticut) for their generous gift of the Phd1fl/fl, Phd2fl/fl and Phd3fl/fl mice. We thank J. Boucher (Joslin Diabetes Center, Boston, MA) for sharing qPCR primer sequences and his critical reading of the manuscript. We thank S. Biddinger (Boston Children's Hospital, Boston, MA) for providing the Fao hepatoma cells. C.M.T. was supported by Radiological Society of North America Research Resident grants 1018 and 1111. E.C.F. and E.L.L. were supported by US National Cancer Institute Training Grant CA121940. C.W. was supported by a training grant from the Canadian Institutes of Health and Research. A.N.D. was supported by a T32 training grant in Comparative Animal Medicine at Stanford University. A.J.K. was supported by grant P20 GM104936 from the US National Institute of General Medical Sciences (NIGMS). Fellowship support was from the NIGMS Stanford Medical Scientist Training Program grant T32 GM007365 (K.W.), Stanford Medical Science Training Program funding (K.W. and L.M.M.), Molecular and Cellular Immunobiology Program training grant 5T32AI07290 (L.M.M.), and US National Institutes of Health (NIH) R01HL074267, R01NS064517 and R01CA158528 (C.J.K.). A.J.G. was supported by NIH grants CA67166 and CA88480 and the Sidney Frank Foundation.

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Authors and Affiliations

  1. Division of Radiation and Cancer Biology, Department of Radiation Oncology, Center for Clinical Sciences Research, Stanford, California, USA

    Cullen M Taniguchi, Elizabeth C Finger, Colleen Wu, Anh N Diep, Edward L LaGory & Amato J Giaccia

  2. Department of Obstetrics and Gynecology, University of Kansas Medical Center, Kansas City, Kansas, USA

    Adam J Krieg

  3. Division of Hematology, Stanford University, Stanford, California, USA

    Kevin Wei, Lisa M McGinnis, Jenny Yuan & Calvin J Kuo

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  1. Cullen M Taniguchi
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Contributions

C.M.T., E.C.F., A.J.K., E.L.L., K.W. and L.M.M. designed and performed experiments and analyzed data. C.W. and A.N.D. generated the knockout animals and contributed to design of all animal experiments. J.Y. and C.J.K. generated and purified the adenoviruses and contributed to experimental design of all adenovirus experiments. C.M.T. and A.J.G. wrote the manuscript and oversaw all aspects of this project.

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Correspondence to Amato J Giaccia.

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Taniguchi, C., Finger, E., Krieg, A. et al. Cross-talk between hypoxia and insulin signaling through Phd3 regulates hepatic glucose and lipid metabolism and ameliorates diabetes. Nat Med 19, 1325–1330 (2013). https://doi.org/10.1038/nm.3294

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