The UDP-sugar-sensing P2Y(14) receptor promotes Rho-mediated signaling and chemotaxis in human neutrophils.

The G(i)-coupled P2Y(14) receptor (P2Y(14)-R) is potently activated by UDP-sugars and UDP. Although P2Y(14)-R mRNA is prominently expressed in circulating neutrophils, the signaling pathways and functional responses associated with this receptor are undefined. In this study, we illustrate that incubation of isolated human neutrophils with UDP-glucose resulted in cytoskeleton rearrangement, change of cell shape, and enhanced cell migration. We also demonstrate that UDP-glucose promotes rapid, robust, and concentration-dependent activation of RhoA in these cells. Ecto-nucleotidases expressed on neutrophils rapidly hydrolyzed extracellular ATP, but incubation with UDP-glucose for up to 1 h resulted in negligible metabolism of the nucleotide-sugar. HL60 human promyelocytic leukemia cells do not express the P2Y(14)-R, but neutrophil differentiation of HL60 cells with DMSO resulted in markedly enhanced P2Y(14)-R expression. Accordingly, UDP-glucose, UDP-galactose, and UDP-N-acetylglucosamine promoted Rho activation in differentiated but not in undifferentiated HL60 cells. Stable expression of recombinant human P2Y(14)-R conferred UDP-sugar-promoted responses to undifferentiated HL60 cells. UDP-glucose-promoted RhoA activation also was accompanied by enhanced cell migration in differentiated HL60 cells, and these responses were blocked by Rho kinase inhibitors. These results support the notion that UDP-glucose is a stable and potent proinflammatory mediator that promotes P2Y(14)-R-mediated neutrophil motility via Rho/Rho kinase activation.

[1]  S. O’Grady.  Purinergic signaling and immune cell chemotaxis. Focus on "the UDP-sugar-sensing P2Y14 receptor promotes Rho-mediated signaling and chemotaxis in human neutrophils". , 2012, American journal of physiology. Cell physiology.

[2]  P. Insel,et al.  New insights regarding the regulation of chemotaxis by nucleotides, adenosine, and their receptors , 2012, Purinergic Signalling.

[3]  P. Kubes,et al.  Sensing sterile injury: opportunities for pharmacological control. , 2011, Pharmacology & therapeutics.

[4]  Seiko F. Okada,et al.  Coupled nucleotide and mucin hypersecretion from goblet-cell metaplastic human airway epithelium. , 2011, American journal of respiratory cell and molecular biology.

[5]  J. Sévigny,et al.  NTPDase1 Controls IL-8 Production by Human Neutrophils , 2011, The Journal of Immunology.

[6]  Seiko F. Okada,et al.  Rho Signaling Regulates Pannexin 1-mediated ATP Release from Airway Epithelia* , 2011, The Journal of Biological Chemistry.

[7]  G. Danuser,et al.  Protein Kinase A Governs a RhoA-RhoGDI Protrusion-Retraction Pacemaker in Migrating Cells , 2011, Nature Cell Biology.

[8]  I. von Kügelgen,et al.  Molecular pharmacology, physiology, and structure of the P2Y receptors. , 2011, Advances in pharmacology.

[9]  E. Lazarowski,et al.  Molecular mechanisms of purine and pyrimidine nucleotide release. , 2011, Advances in pharmacology.

[10]  P. Kubes,et al.  Intravascular Danger Signals Guide Neutrophils to Sites of Sterile Inflammation , 2010, Science.

[11]  W. Junger,et al.  Purinergic Signaling: A Fundamental Mechanism in Neutrophil Activation , 2010, Science Signaling.

[12]  T. K. Harden,et al.  Signalling and pharmacological properties of the P2Y14 receptor , 2010, Acta physiologica.

[13]  K. Jacobson,et al.  UDP‐glucose acting at P2Y14 receptors is a mediator of mast cell degranulation , 2010, Biochemical pharmacology.

[14]  E. Lazarowski.  Quantification of extracellular UDP-galactose. , 2010, Analytical biochemistry.

[15]  K. Jacobson,et al.  Quantification of Gi-Mediated Inhibition of Adenylyl Cyclase Activity Reveals That UDP Is a Potent Agonist of the Human P2Y14 Receptor , 2009, Molecular Pharmacology.

[16]  Gaudenz Danuser,et al.  Coordination of Rho GTPase activities during cell protrusion , 2009, Nature.

[17]  R. Marée,et al.  P2X1 Ion Channels Promote Neutrophil Chemotaxis through Rho Kinase Activation , 2009 .

[18]  T. K. Harden,et al.  Gi-Dependent Cell Signaling Responses of the Human P2Y14 Receptor in Model Cell Systems , 2009, Journal of Pharmacology and Experimental Therapeutics.

[19]  T. Nagashima,et al.  The UDP-Glucose Receptor P2RY14 Triggers Innate Mucosal Immunity in the Female Reproductive Tract by Inducing IL-81 , 2009, The Journal of Immunology.

[20]  E. Lazarowski,et al.  Endoplasmic Reticulum/Golgi Nucleotide Sugar Transporters Contribute to the Cellular Release of UDP-sugar Signaling Molecules* , 2009, Journal of Biological Chemistry.

[21]  P. Insel,et al.  Ecto-nucleoside Triphosphate Diphosphohydrolase 1 (E-NTPDase1/CD39) Regulates Neutrophil Chemotaxis by Hydrolyzing Released ATP to Adenosine* , 2008, Journal of Biological Chemistry.

[22]  A. Ault,et al.  Similarities between UDP-glucose and adenine nucleotide release in yeast: involvement of the secretory pathway. , 2008, Biochemistry.

[23]  K. Jacobson,et al.  UDP Is a Competitive Antagonist at the Human P2Y14 Receptor , 2008, Journal of Pharmacology and Experimental Therapeutics.

[24]  E. Lazarowski,et al.  Thrombin‐promoted release of UDP‐glucose from human astrocytoma cells , 2008, British journal of pharmacology.

[25]  G. Dubyak,et al.  The inflammatory effects of UDP-glucose in N9 microglia are not mediated by P2Y14 receptor activation , 2008, Purinergic Signalling Purinergic Signalling.

[26]  H. Bourne,et al.  PDZRhoGEF and myosin II localize RhoA activity to the back of polarizing neutrophil-like cells , 2007, The Journal of cell biology.

[27]  A. Surprenant,et al.  Inhibition of Neutrophil Apoptosis by ATP Is Mediated by the P2Y11 Receptor1 , 2007, The Journal of Immunology.

[28]  Seiko F. Okada,et al.  Coordinated release of nucleotides and mucin from human airway epithelial Calu‐3 cells , 2007, The Journal of physiology.

[29]  J. V. van Buul,et al.  VEGF-induced Rac1 activation in endothelial cells is regulated by the guanine nucleotide exchange factor Vav2. , 2007, Experimental cell research.

[30]  M. J. Broekman,et al.  CD39/NTPDase-1 activity and expression in normal leukocytes. , 2007, Thrombosis research.

[31]  Linda Yip,et al.  ATP Release Guides Neutrophil Chemotaxis via P2Y2 and A3 Receptors , 2006, Science.

[32]  J. Dickenson,et al.  Functional expression of the P2Y14 receptor in human neutrophils. , 2006, European journal of pharmacology.

[33]  A. Mollica,et al.  Signal transduction pathways triggered by selective formylpeptide analogues in human neutrophils. , 2006, European journal of pharmacology.

[34]  F. Di Virgilio,et al.  The P2Y14 receptor of airway epithelial cells: coupling to intracellular Ca2+ and IL-8 secretion. , 2005, American journal of respiratory cell and molecular biology.

[35]  J. Cherfils,et al.  Uncoupling of Inhibitory and Shuttling Functions of Rho GDP Dissociation Inhibitors* , 2005, Journal of Biological Chemistry.

[36]  R. Ye,et al.  A Rho Exchange Factor Mediates fMet-Leu-Phe-induced NF-κB Activation in Human Peripheral Blood Monocytes* , 2004, Journal of Biological Chemistry.

[37]  V. Niggli.  Signaling to migration in neutrophils: importance of localized pathways. , 2003, The international journal of biochemistry & cell biology.

[38]  P. Emson,et al.  GPR105, a novel Gi/o-coupled UDP-glucose receptor expressed on brain glia and peripheral immune cells, is regulated by immunologic challenge: possible role in neuroimmune function. , 2003, Brain research. Molecular brain research.

[39]  A. Platt,et al.  Human Immature Monocyte-Derived Dendritic Cells Express the G Protein-Coupled Receptor GPR105 (KIAA0001, P2Y14) and Increase Intracellular Calcium in Response to its Agonist, Uridine Diphosphoglucose , 2003, The Journal of Immunology.

[40]  Jingsong Xu,et al.  Divergent Signals and Cytoskeletal Assemblies Regulate Self-Organizing Polarity in Neutrophils , 2003, Cell.

[41]  D. Haber,et al.  P2Y-like receptor, GPR105 (P2Y14), identifies and mediates chemotaxis of bone-marrow hematopoietic stem cells. , 2003, Genes & development.

[42]  R. Boucher,et al.  Release of cellular UDP-glucose as a potential extracellular signaling molecule. , 2003, Molecular pharmacology.

[43]  V. V. van Hinsbergh,et al.  Involvement of RhoA/Rho Kinase Signaling in VEGF-Induced Endothelial Cell Migration and Angiogenesis In Vitro , 2003, Arteriosclerosis, thrombosis, and vascular biology.

[44]  D. Mukhopadhyay,et al.  KDR Stimulates Endothelial Cell Migration through Heterotrimeric G Protein Gq/11-mediated Activation of a Small GTPase RhoA* , 2002, The Journal of Biological Chemistry.

[45]  P. Emson,et al.  Cloning, pharmacology, and tissue distribution of G-protein-coupled receptor GPR105 (KIAA0001) rodent orthologs. , 2001, Genomics.

[46]  P. Hordijk,et al.  Activation of Rhoa and ROCK are essential for detachment of migrating leukocytes. , 2001, Molecular biology of the cell.

[47]  R. Boucher,et al.  Constitutive Release of ATP and Evidence for Major Contribution of Ecto-nucleotide Pyrophosphatase and Nucleoside Diphosphokinase to Extracellular Nucleotide Concentrations* , 2000, The Journal of Biological Chemistry.

[48]  J. Chambers,et al.  A G Protein-coupled Receptor for UDP-glucose* , 2000, The Journal of Biological Chemistry.

[49]  T. K. Harden,et al.  Quantitation of extracellular UTP using a sensitive enzymatic assay , 1999, British journal of pharmacology.

[50]  V. Niggli.  Rho‐kinase in human neutrophils: a role in signalling for myosin light chain phosphorylation and cell migration , 1999, FEBS letters.

[51]  G. Dubyak,et al.  The P2X1 receptor, an adenosine triphosphate-gated cation channel, is expressed in human platelets but not in human blood leukocytes. , 1998, Blood.

[52]  F. Sistare,et al.  Distribution of P2Y receptor subtypes on haematopoietic cells , 1998, British journal of pharmacology.

[53]  Robert Kay,et al.  Lfc and Lsc Oncoproteins Represent Two New Guanine Nucleotide Exchange Factors for the Rho GTP-binding Protein* , 1996, The Journal of Biological Chemistry.

[54]  J. Boucheron,et al.  P2U Agonists Induce Chemotaxis and Actin Polymerization in Human Neutrophils and Differentiated HL60 Cells* , 1996, The Journal of Biological Chemistry.

[55]  A. Surprenant,et al.  P2X1 receptor activation in HL60 cells. , 1996, Blood.

[56]  David A. Williams,et al.  Novel human oncogene lbc detected by transfection with distinct homology regions to signal transduction products. , 1994, Oncogene.

[57]  Anika Niambi Al-Shura.  Leukocytes. , 1994, Current opinion in hematology.

[58]  E. Lazarowski,et al.  Aggregation of human neutrophils by heparin. , 1986, Thrombosis research.