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

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.

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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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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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The authors declare no competing financial interests.

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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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