Dysfunction of the PI3 kinase/Rap1/integrin α(IIb)β(3) pathway underlies ex vivo platelet hypoactivity in essential thrombocythemia

Blood. 2013 Feb 14;121(7):1209-19. doi: 10.1182/blood-2012-05-431288. Epub 2012 Dec 13.

Abstract

Patients with myeloproliferative disorders (MPDs), such as essential thrombocythemia (ET) have increased risk of thrombosis and bleeding, which are major sources of morbidity and mortality. Most MPD patients have a gain of function mutation in Janus kinase 2 (JAK2V617F), but little is known how JAK2V617F affects platelet function. Here, we demonstrate that platelets from ET patients have impaired SFLLRN-mediated fibrinogen binding and have lost the potentiating effect of thrombopoietin (which couples to JAK2) on this pathway. In contrast, SFLLRN-mediated P-selectin expression, ATP secretion, phosphorylation of the PKC substrate pleckstrin, and Ca(2+) mobilization were unaffected in JAK2V617F positive platelets. In addition, thrombopoietin-mediated JAK2 phosphorylation was unchanged, suggesting that signaling pathways activated downstream of JAK2 are impaired. Indeed, we found that platelets from JAK2V617F positive ET patients have significantly reduced phosphorylation of the PI3 kinase substrate Akt, and have reduced activation of Rap1 in response to thrombopoietin, IGF-1,ADP, SFLLRN, and thrombin. This effect was independent of Giα P2Y12 purinergic receptor function as ADP-mediated inhibition of VASP phosphorylation was unchanged. These results demonstrate that the PI3 kinase/Rap1 pathway is intrinsically impaired in platelets from JAK2V617F-positive ET patients, resulting in diminished thrombin and thrombopoietin-mediated integrin α(IIb)β(3) activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Amino Acid Substitution
  • Blood Platelets / drug effects
  • Blood Platelets / physiology*
  • Case-Control Studies
  • Female
  • Fibrinogen / metabolism
  • Humans
  • Janus Kinase 2 / antagonists & inhibitors
  • Janus Kinase 2 / blood
  • Janus Kinase 2 / genetics
  • Male
  • Middle Aged
  • Mutation, Missense
  • Peptide Fragments / pharmacology
  • Phosphatidylinositol 3-Kinases / blood*
  • Phosphorylation
  • Platelet Activation / drug effects
  • Platelet Activation / genetics
  • Platelet Activation / physiology*
  • Platelet Glycoprotein GPIIb-IIIa Complex / metabolism*
  • Shelterin Complex
  • Signal Transduction / drug effects
  • Telomere-Binding Proteins / blood*
  • Thrombin / pharmacology
  • Thrombocythemia, Essential / blood*
  • Thrombocythemia, Essential / genetics
  • Thrombopoietin / pharmacology

Substances

  • Peptide Fragments
  • Platelet Glycoprotein GPIIb-IIIa Complex
  • Shelterin Complex
  • TERF2IP protein, human
  • Telomere-Binding Proteins
  • thrombin receptor peptide (42-47)
  • Fibrinogen
  • Thrombopoietin
  • JAK2 protein, human
  • Janus Kinase 2
  • Thrombin