Prolyl hydroxylase 2 inactivation enhances glycogen storage and promotes excessive neutrophilic responses

Fully activated innate immune cells are required for effective responses to infection, but their prompt deactivation and removal are essential for limiting tissue damage. Here, we have identified a critical role for the prolyl hydroxylase enzyme Phd2 in maintaining the balance between appropriate, predominantly neutrophil-mediated pathogen clearance and resolution of the innate immune response. We demonstrate that myeloid-specific loss of Phd2 resulted in an exaggerated inflammatory response to Streptococcus pneumonia, with increases in neutrophil motility, functional capacity, and survival. These enhanced neutrophil responses were dependent upon increases in glycolytic flux and glycogen stores. Systemic administration of a HIF–prolyl hydroxylase inhibitor replicated the Phd2-deficient phenotype of delayed inflammation resolution. Together, these data identify Phd2 as the dominant HIF-hydroxylase in neutrophils under normoxic conditions and link intrinsic regulation of glycolysis and glycogen stores to the resolution of neutrophil-mediated inflammatory responses. These results demonstrate the therapeutic potential of targeting metabolic pathways in the treatment of inflammatory disease.

[1]  A. Shah,et al.  PHD2 Is a Regulator for Glycolytic Reprogramming in Macrophages , 2016, Molecular and Cellular Biology.

[2]  P. Carmeliet,et al.  Mutations in succinate dehydrogenase B (SDHB) enhance neutrophil survival independent of HIF-1α expression. , 2016, Blood.

[3]  W. Junger,et al.  mTOR and differential activation of mitochondria orchestrate neutrophil chemotaxis , 2015, The Journal of cell biology.

[4]  W. Kaelin,et al.  Inhibition of the oxygen sensor PHD2 in the liver improves survival in lactic acidosis by activating the Cori cycle , 2015, Proceedings of the National Academy of Sciences.

[5]  P. Ratcliffe,et al.  Potent and Selective Triazole-Based Inhibitors of the Hypoxia-Inducible Factor Prolyl-Hydroxylases with Activity in the Murine Brain , 2015, PloS one.

[6]  Z. Granot,et al.  MET is required for the recruitment of anti-tumoural neutrophils , 2015, Nature.

[7]  Abhishek K. Jha,et al.  Network integration of parallel metabolic and transcriptional data reveals metabolic modules that regulate macrophage polarization. , 2015, Immunity.

[8]  P. Carmeliet,et al.  Fatty acid carbon is essential for dNTP synthesis in endothelial cells , 2015, Nature.

[9]  Susan R. Quinn,et al.  Pyruvate kinase M2 regulates Hif-1α activity and IL-1β induction and is a critical determinant of the warburg effect in LPS-activated macrophages. , 2015, Cell metabolism.

[10]  Susan R. Quinn,et al.  Pyruvate Kinase M2 Regulates Hif-1α Activity and IL-1β Induction and Is a Critical Determinant of the Warburg Effect in LPS-Activated Macrophages. , 2015, Cell metabolism.

[11]  J. Keldenich,et al.  Mimicking Hypoxia to Treat Anemia: HIF-Stabilizer BAY 85-3934 (Molidustat) Stimulates Erythropoietin Production without Hypertensive Effects , 2014, PloS one.

[12]  J. Chou,et al.  PHAGOCYTES , GRANULOCYTES , AND MYELOPOIESIS Molecular mechanisms of neutrophil dysfunction in glycogen storage disease type Ib , 2014 .

[13]  C. Schofield,et al.  Non-enzymatic chemistry enables 2-hydroxyglutarate-mediated activation of 2-oxoglutarate oxygenases , 2014, Nature Communications.

[14]  F. Formenti,et al.  Hypoxia-inducible factor 2α regulates key neutrophil functions in humans, mice, and zebrafish. , 2014, Blood.

[15]  P. Carmeliet,et al.  Role of PFKFB3-Driven Glycolysis in Vessel Sprouting , 2013, Cell.

[16]  Liang Zheng,et al.  Succinate is an inflammatory signal that induces IL-1β through HIF-1α , 2013, Nature.

[17]  P. Carmeliet,et al.  Emerging novel functions of the oxygen-sensing prolyl hydroxylase domain enzymes. , 2013, Trends in biochemical sciences.

[18]  R. Verhaak,et al.  Transformation by the R Enantiomer of 2-Hydroxyglutarate Linked to EglN Activation , 2012, Nature.

[19]  K. Alitalo,et al.  Macrophage skewing by Phd2 haplodeficiency prevents ischaemia by inducing arteriogenesis , 2011, Nature.

[20]  Matthew K. Knabel,et al.  Pyruvate Kinase M2 Is a PHD3-Stimulated Coactivator for Hypoxia-Inducible Factor 1 , 2011, Cell.

[21]  R. Klose,et al.  The oncometabolite 2‐hydroxyglutarate inhibits histone lysine demethylases , 2011, EMBO reports.

[22]  Nicola Zamboni,et al.  The oxygen sensor PHD3 limits glycolysis under hypoxia via direct binding to pyruvate kinase , 2011, Cell Research.

[23]  P. Carmeliet,et al.  Prolyl hydroxylase 3 (PHD3) is essential for hypoxic regulation of neutrophilic inflammation in humans and mice. , 2011, The Journal of clinical investigation.

[24]  P. Carmeliet,et al.  Absence of placental growth factor blocks dextran sodium sulfate-induced colonic mucosal angiogenesis, increases mucosal hypoxia and aggravates acute colonic injury , 2010, Laboratory Investigation.

[25]  Frederik De Smet,et al.  Heterozygous Deficiency of PHD2 Restores Tumor Oxygenation and Inhibits Metastasis via Endothelial Normalization , 2009, Cell.

[26]  F. Galateau-Sallé,et al.  PHD2 mutation and congenital erythrocytosis with paraganglioma. , 2008, The New England journal of medicine.

[27]  W. Kaelin,et al.  Oxygen sensing by metazoans: the central role of the HIF hydroxylase pathway. , 2008, Molecular cell.

[28]  David A Ferrick,et al.  Advances in measuring cellular bioenergetics using extracellular flux. , 2008, Drug discovery today.

[29]  M. McMullin,et al.  A novel erythrocytosis-associated PHD2 mutation suggests the location of a HIF binding groove. , 2007, Blood.

[30]  J. Chou,et al.  Impaired neutrophil activity and increased susceptibility to bacterial infection in mice lacking glucose-6-phosphatase-beta. , 2007, The Journal of clinical investigation.

[31]  K. Takeda,et al.  Placental but Not Heart Defects Are Associated with Elevated Hypoxia-Inducible Factor α Levels in Mice Lacking Prolyl Hydroxylase Domain Protein 2 , 2006, Molecular and Cellular Biology.

[32]  M. McMullin,et al.  A family with erythrocytosis establishes a role for prolyl hydroxylase domain protein 2 in oxygen homeostasis. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[33]  C. Print,et al.  Hypoxia-induced neutrophil survival is mediated by HIF-1α–dependent NF-κB activity , 2005, The Journal of experimental medicine.

[34]  A. Harris,et al.  Differential Function of the Prolyl Hydroxylases PHD1, PHD2, and PHD3 in the Regulation of Hypoxia-inducible Factor* , 2004, Journal of Biological Chemistry.

[35]  Christopher J. Schofield,et al.  Oxygen sensing by HIF hydroxylases , 2004, Nature Reviews Molecular Cell Biology.

[36]  P. Ince,et al.  Alveolar Macrophage Apoptosis Contributes to Pneumococcal Clearance in a Resolving Model of Pulmonary Infection 1 , 2003, The Journal of Immunology.

[37]  J. Pouysségur,et al.  HIF prolyl‐hydroxylase 2 is the key oxygen sensor setting low steady‐state levels of HIF‐1α in normoxia , 2003, The EMBO journal.

[38]  Z. Estrov,et al.  Granulocyte-macrophage colony-stimulating factor (GM-CSF) induces antiapoptotic and proapoptotic signals in acute myeloid leukemia. , 2003, Blood.

[39]  R. Jaenisch,et al.  HIF-1α Is Essential for Myeloid Cell-Mediated Inflammation , 2003, Cell.

[40]  R. Jaenisch,et al.  HIF-1alpha is essential for myeloid cell-mediated inflammation. , 2003, Cell.

[41]  V. Ridger,et al.  Caspase-1-Deficient Mice Have Delayed Neutrophil Apoptosis and a Prolonged Inflammatory Response to Lipopolysaccharide-Induced Acute Lung Injury1 , 2002, The Journal of Immunology.

[42]  Michael I. Wilson,et al.  C. elegans EGL-9 and Mammalian Homologs Define a Family of Dioxygenases that Regulate HIF by Prolyl Hydroxylation , 2001, Cell.

[43]  H. Brunengraber,et al.  Correction of 13C mass isotopomer distributions for natural stable isotope abundance. , 1996, Journal of mass spectrometry : JMS.

[44]  M. Karnovsky,et al.  Glycogen accumulation in polymorphonuclear leukocytes, and other intracellular alterations that occur during inflammation , 1982, The Journal of cell biology.

[45]  G. Ackerman.  HISTOCHEMICAL DIFFERENTIATION DURING NEUTROPHIL DEVELOPMENT AND MATURATION * , 1964, Annals of the New York Academy of Sciences.

[46]  P. Levene,et al.  THE ACTION OF LEUCOCYTES ON GLUCOSE , 1912 .