Opposing Roles of GSK3α and GSK3β Phosphorylation in Platelet Function and Thrombosis

Int J Mol Sci. 2021 Sep 30;22(19):10656. doi: 10.3390/ijms221910656.

Abstract

One of the mechanisms by which PI3 kinase can regulate platelet function is through phosphorylation of downstream substrates, including glycogen synthase kinase-3 (GSK3)α and GSK3β. Platelet activation results in the phosphorylation of an N-terminal serine residue in GSK3α (Ser21) and GSK3β (Ser9), which competitively inhibits substrate phosphorylation. However, the role of phosphorylation of these paralogs is still largely unknown. Here, we employed GSK3α/β phosphorylation-resistant mouse models to explore the role of this inhibitory phosphorylation in regulating platelet activation. Expression of phosphorylation-resistant GSK3α/β reduced thrombin-mediated platelet aggregation, integrin αIIbβ3 activation, and α-granule secretion, whereas platelet responses to the GPVI agonist collagen-related peptide (CRP-XL) were significantly enhanced. GSK3 single knock-in lines revealed that this divergence is due to differential roles of GSK3α and GSK3β phosphorylation in regulating platelet function. Expression of phosphorylation-resistant GSK3α resulted in enhanced GPVI-mediated platelet activation, whereas expression of phosphorylation-resistant GSK3β resulted in a reduction in PAR-mediated platelet activation and impaired in vitro thrombus formation under flow. Interestingly, the latter was normalised in double GSK3α/β KI mice, indicating that GSK3α KI can compensate for the impairment in thrombosis caused by GSK3β KI. In conclusion, our data indicate that GSK3α and GSK3β have differential roles in regulating platelet function.

Keywords: Akt; GSK3; P-selectin expression; PI3 kinase; integrin activation; knock-in mouse models; p110β; phosphorylation; thrombosis.

MeSH terms

  • Animals
  • Blood Donors
  • Blood Platelets / metabolism*
  • Cells, Cultured
  • Disease Models, Animal
  • Gene Knock-In Techniques
  • Glycogen Synthase Kinase 3 / genetics
  • Glycogen Synthase Kinase 3 / metabolism*
  • Glycogen Synthase Kinase 3 beta / genetics
  • Glycogen Synthase Kinase 3 beta / metabolism*
  • Humans
  • Integrins / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphorylation / genetics
  • Platelet Activation / genetics*
  • Platelet Aggregation / genetics*
  • Proto-Oncogene Proteins c-akt / metabolism
  • Signal Transduction / genetics*
  • Thrombin / metabolism
  • Thrombosis / genetics
  • Thrombosis / metabolism*

Substances

  • Integrins
  • GSK3B protein, human
  • Glycogen Synthase Kinase 3 beta
  • Gsk3b protein, mouse
  • Proto-Oncogene Proteins c-akt
  • Glycogen Synthase Kinase 3
  • glycogen synthase kinase 3 alpha
  • Thrombin