Critical roles for the phosphatidylinositide 3-kinase isoforms p110β and p110γ in thrombopoietin-mediated priming of platelet function

• Published in: Scientific reports 2019-02, Vol.9 (1), p.1468-1468, Article 1468
• Main Authors: Moore, Samantha F., Smith, Nina R., Blair, Thomas A., Durrant, Tom N., Hers, Ingeborg
• Format: Article
• Language: English
• Subjects: 1-Phosphatidylinositol 3-kinase; 631/80/458/1733; 631/80/86/2364; 64/110; 692/699/75; 82/29; 96/31; 96/95; AKT protein; Animals; Blood Platelets - drug effects; Blood Platelets - metabolism; Class I Phosphatidylinositol 3-Kinases - metabolism; Class Ib Phosphatidylinositol 3-Kinase - metabolism; Extracellular signal-regulated kinase; Guanosine triphosphatases; Humanities and Social Sciences; Humans; Isoforms; Janus kinase 2; Lipid kinase; Mice; multidisciplinary; Phosphorylation; Platelet Activation - drug effects; Platelets; Protein-tyrosine kinase; Proto-Oncogene Proteins c-akt - metabolism; rap1 GTP-Binding Proteins - metabolism; Rap1 protein; Science; Science (multidisciplinary); Secretion; Synergistic effect; Thrombopoietin; Thrombopoietin - pharmacology; Thromboxanes - metabolism; Thromboxanes - pharmacology
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-018-37012-9

Thrombopoietin (TPO) enhances platelet activation through activation of the tyrosine kinase; JAK2 and the lipid kinase phosphatidylinositide 3-kinase (PI3K). The aim of our study was to identify the PI3K isoforms involved in mediating the effect of TPO on platelet function and elucidate the underlying mechanism. We found that p110β plays an essential role in TPO-mediated (i) priming of protease-activated receptor (PAR)-mediated integrin α IIb β 3 activation and α-granule secretion, (ii) synergistic enhancement of PAR-mediated activation of the small GTPase RAP1, a regulator of integrin activation and (iii) phosphorylation of the PI3K effector Akt. More importantly, the synergistic effect of TPO on phosphorylation of extracellular-regulated kinase (ERK1/2) and thromboxane (TxA 2 ) synthesis was dependent on both p110β and p110γ. p110β inhibition/deletion, or inhibition of p110γ, resulted in a partial reduction, whereas inhibiting both p110β and p110γ completely prevented the synergistic effect of TPO on ERK1/2 phosphorylation and TxA 2 synthesis. The latter was ablated by inhibition of MEK, but not p38, confirming a role for ERK1/2 in regulating TPO-mediated increases in TxA 2 synthesis. In conclusion, the synergistic effect of TPO on RAP1 and integrin activation is largely mediated by p110β, whereas p110β and p110γ contribute to the effect of TPO on ERK1/2 phosphorylation and TxA 2 formation.