title: Shear stress modulates endothelial KLF2 through activation of P2X4.
abstract: Vascular endothelial cells that are in direct contact with blood flow are exposed to fluid shear stress and regulate vascular homeostasis. Studies report endothelial cells to release ATP in response to shear stress that in turn modulates cellular functions via P2 receptors with P2X4 mediating shear stress-induced calcium signaling and vasodilation. A recent study shows that a loss-of-function polymorphism in the human P2X4 resulting in a Tyr315>Cys variant is associated with increased pulse pressure and impaired endothelial vasodilation. Although the importance of shear stress-induced Krüppel-like factor 2 (KLF2) expression in atheroprotection is well studied, whether ATP regulates KLF2 remains unanswered and is the objective of this study. Using an in vitro model, we show that in human umbilical vein endothelial cells (HUVECs), apyrase decreased shear stress-induced KLF2, KLF4, and NOS3 expression but not that of NFE2L2. Exposure of HUVECs either to shear stress or ATPγS under static conditions increased KLF2 in a P2X4-dependent manner as was evident with both the receptor antagonist and siRNA knockdown. Furthermore, transient transfection of static cultures of human endothelial cells with the Tyr315>Cys mutant P2X4 construct blocked ATP-induced KLF2 expression. Also, P2X4 mediated the shear stress-induced phosphorylation of extracellular regulated kinase-5, a known regulator of KLF2. This study demonstrates a major physiological finding that the shear-induced effects on endothelial KLF2 axis are in part dependent on ATP release and P2X4, a previously unidentified mechanism.
authors.affiliation: Department of Cardiology, Clinical Sciences, Lund University, Lund, Sweden, Ramasri.Sathanoori@med.lu.se.
authors.forename: R
authors.initials: R
authors.lastname: Sathanoori
authors.forename: F
authors.initials: F
authors.lastname: Rosi
authors.forename: B J
authors.initials: BJ
authors.lastname: Gu
authors.forename: J S
authors.initials: JS
authors.lastname: Wiley
authors.forename: C E
authors.initials: CE
authors.lastname: Müller
authors.forename: B
authors.initials: B
authors.lastname: Olde
authors.forename: D
authors.initials: D
authors.lastname: Erlinge
mesh_terms: D002478:Cells, Cultured; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D058920:Purinergic P2X Receptor Antagonists / Q000494:pharmacology; D034741:RNA, Small Interfering; D058848:Receptors, Purinergic P2X4 / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013314:Stress, Mechanical
chemical_list: C096161:KLF2 protein, human; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D058920:Purinergic P2X Receptor Antagonists; D034741:RNA, Small Interfering; D058848:Receptors, Purinergic P2X4; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III