Selective induction of endothelial P2Y6 nucleotide receptor promotes vascular inflammation

TY - JOUR

T1 - Selective induction of endothelial P2Y6 nucleotide receptor promotes vascular inflammation

AU - Riegel, Ann-Kathrin

AU - Faigle, Marion

AU - Zug, Stephanie

AU - Rosenberger, Peter

AU - Robaye, Bernard

AU - Boeynaems, Jean-Marie

AU - Idzko, Marco

AU - Eltzschig, Holger K

PY - 2011/2/24

Y1 - 2011/2/24

N2 - During a systemic inflammatory response endothelial-expressed surface molecules have been strongly implicated in orchestrating immune responses. Previous studies have shown enhanced extracellular nucleotide release during acute inflammatory conditions. Therefore, we hypothesized that endothelial nucleotide receptors could play a role in vascular inflammation. To address this hypothesis, we performed screening experiments and exposed human microvascular endothelia to inflammatory stimuli, followed by measurements of P2Y or P2X transcriptional responses. These studies showed a selective induction of the P2Y(6) receptor (> 4-fold at 24 hours). Moreover, studies that used real-time reverse transcription-polymerase chain reaction, Western blot analysis, or immunofluorescence confirmed time- and dose-dependent induction of P2Y(6) with tumor necrosis factor α or Lipopolysaccharide (LPS) stimulation in vitro and in vivo. Studies that used MRS 2578 as P2Y(6) receptor antagonist showed attenuated nuclear factor κB reporter activity and proinflammatory gene expression in human microvascular endothelial cells in vitro. Moreover, pharmacologic or genetic in vivo studies showed attenuated inflammatory responses in P2Y(6)(-/-) mice or after P2Y(6) antagonist treatment during LPS-induced vascular inflammation. These studies show an important contribution of P2Y(6) signaling in enhancing vascular inflammation during systemic LPS challenge and implicate the P2Y(6) receptor as a therapeutic target during systemic inflammatory responses.

AB - During a systemic inflammatory response endothelial-expressed surface molecules have been strongly implicated in orchestrating immune responses. Previous studies have shown enhanced extracellular nucleotide release during acute inflammatory conditions. Therefore, we hypothesized that endothelial nucleotide receptors could play a role in vascular inflammation. To address this hypothesis, we performed screening experiments and exposed human microvascular endothelia to inflammatory stimuli, followed by measurements of P2Y or P2X transcriptional responses. These studies showed a selective induction of the P2Y(6) receptor (> 4-fold at 24 hours). Moreover, studies that used real-time reverse transcription-polymerase chain reaction, Western blot analysis, or immunofluorescence confirmed time- and dose-dependent induction of P2Y(6) with tumor necrosis factor α or Lipopolysaccharide (LPS) stimulation in vitro and in vivo. Studies that used MRS 2578 as P2Y(6) receptor antagonist showed attenuated nuclear factor κB reporter activity and proinflammatory gene expression in human microvascular endothelial cells in vitro. Moreover, pharmacologic or genetic in vivo studies showed attenuated inflammatory responses in P2Y(6)(-/-) mice or after P2Y(6) antagonist treatment during LPS-induced vascular inflammation. These studies show an important contribution of P2Y(6) signaling in enhancing vascular inflammation during systemic LPS challenge and implicate the P2Y(6) receptor as a therapeutic target during systemic inflammatory responses.

KW - Animals

KW - Endothelial Cells

KW - Endothelium, Vascular

KW - Gene Expression Profiling

KW - Humans

KW - Inflammation

KW - Lipopolysaccharides

KW - Mice

KW - Mice, Knockout

KW - Receptors, Purinergic P2

KW - Receptors, Purinergic P2X

KW - Receptors, Purinergic P2Y

KW - Transcriptional Activation

KW - Journal Article

KW - Research Support, N.I.H., Extramural

U2 - 10.1182/blood-2010-10-313957

DO - 10.1182/blood-2010-10-313957

M3 - SCORING: Journal article

C2 - 21173118

VL - 117

SP - 2548

EP - 2555

JO - BLOOD

JF - BLOOD

SN - 0006-4971

IS - 8

ER -