Platelet PI3K Modulates Innate Leukocyte Extravasation during Acid-Induced Acute Lung InflammationFunding Financial support was provided by grants of the Austrian Science Fond (FWF) to A.A. and G.S. (P24978, SFB F54) and of the Austrian National Bank to A.A. (OeNB grant 15961). The authors declare no competing financial interests.
18 January 2019
12 June 2019
01 August 2019 (eFirst)
Introduction Blood platelets are increasingly recognized as modulators of leukocyte effector functions in various pathologies including acute lung injury (ALI). ALI is a life-threatening disease, caused by damage to the alveolar epi- and endothelium. Excessive accumulation of leukocytes leads to severe lung inflammation, resulting in impaired lung function and hypoxemia.
Objective Since leukocyte migration is modulated by activated platelets and phosphatidylinositol 3-kinase (PI3K) signaling is involved in platelet function, we aimed to elucidate the effect of PI3K on platelet-mediated immune responses.
Materials and Methods We generated a mouse model with a platelet-specific deletion of p85α, the most important regulatory subunit of the class IA PI3K, and evaluated platelet function and platelet–leukocyte interactions. Moreover, we analyzed the impact of platelet-specific p85α gene deficiency during sterile peritonitis and acid-induced ALI.
Results In vitro analyses of platelets revealed that lack of p85α led to decreased downstream signaling and diminished expression of surface activation markers, for example, CD62P and CD63, as well as reduced platelet aggregation. Moreover, platelet PI3K essentially mediated direct interactions of platelets with monocytes and neutrophils. In mice, platelet-specific p85α deficiency prevented leukocyte infiltration into the peritoneum and the bronchoalveolar compartment during sterile peritonitis and ALI, respectively. Additionally, the release of the inflammatory cytokine interleukin-12/23 was diminished in platelet p85α-deficient mice during ALI. In contrast to PI3K, neither overexpression nor depletion of platelet phosphatase and tensin homolog, the endogenous antagonist of PI3K, significantly modulated platelet function.
Conclusion Our data indicate a crucial role of platelet PI3K signaling for leukocyte extravasation upon inflammatory stimuli in various diseases models.
Experiments performed by J.B.K.P., W.C.S., M.S., M.M., S.H., G.J.S., B.M., B.B., and H.P.; data analyzed and evaluated by J.B.K.P., W.C.S., M.S., and M.M.; data interpreted by J.B.K.P., W.C.S., M.Z., A.A., and G.S.; study designed by J.B.K.P., W.C.S., A.A., G.S., and S.K.; figures compiled by J.B.K.P., W.C.S., M.S., and A.A.; manuscript written by J.B.K.P., W.C.S., and A.A.; study coordinated and funding acquired by A.A. and G.S. All authors read and approved the final version of the manuscript.
* Both the authors contributed equally to the study.
- 1 Weyrich AS, Zimmerman GA. Platelets: signaling cells in the immune continuum. Trends Immunol 2004; 25 (09) 489-495
- 2 Zuchtriegel G, Uhl B, Puhr-Westerheide D. , et al. Platelets guide leukocytes to their sites of extravasation. PLoS Biol 2016; 14 (05) e1002459
- 3 Weber C. Platelets and chemokines in atherosclerosis: partners in crime. Circ Res 2005; 96 (06) 612-616
- 4 Kurose I, Anderson DC, Miyasaka M. , et al. Molecular determinants of reperfusion-induced leukocyte adhesion and vascular protein leakage. Circ Res 1994; 74 (02) 336-343
- 5 Zarbock A, Müller H, Kuwano Y, Ley K. PSGL-1-dependent myeloid leukocyte activation. J Leukoc Biol 2009; 86 (05) 1119-1124
- 6 Willecke F, Tiwari S, Rupprecht B. , et al. Interruption of classic CD40L-CD40 signalling but not of the novel CD40L-Mac-1 interaction limits arterial neointima formation in mice. Thromb Haemost 2014; 112 (02) 379-389
- 7 Lievens D, Zernecke A, Seijkens T. , et al. Platelet CD40L mediates thrombotic and inflammatory processes in atherosclerosis. Blood 2010; 116 (20) 4317-4327
- 8 Zirlik A, Maier C, Gerdes N. , et al. CD40 ligand mediates inflammation independently of CD40 by interaction with Mac-1. Circulation 2007; 115 (12) 1571-1580
- 9 Badrnya S, Schrottmaier WC, Kral JB. , et al. Platelets mediate oxidized low-density lipoprotein-induced monocyte extravasation and foam cell formation. Arterioscler Thromb Vasc Biol 2014; 34 (03) 571-580
- 10 Lam FW, Burns AR, Smith CW, Rumbaut RE. Platelets enhance neutrophil transendothelial migration via P-selectin glycoprotein ligand-1. Am J Physiol Heart Circ Physiol 2011; 300 (02) H468-H475
- 11 Wetterö J, Tengvall P, Bengtsson T. Platelets stimulated by IgG-coated surfaces bind and activate neutrophils through a selectin-dependent pathway. Biomaterials 2003; 24 (09) 1559-1573
- 12 Huo Y, Schober A, Forlow SB. , et al. Circulating activated platelets exacerbate atherosclerosis in mice deficient in apolipoprotein E. Nat Med 2003; 9 (01) 61-67
- 13 Vardon Bounes F, Mujalli A, Cenac C. , et al. The importance of blood platelet lipid signaling in thrombosis and in sepsis. Adv Biol Regul 2018; 67: 66-73
- 14 Domin J, Waterfield MD. Using structure to define the function of phosphoinositide 3-kinase family members. FEBS Lett 1997; 410 (01) 91-95
- 15 Watanabe N, Nakajima H, Suzuki H. , et al. Functional phenotype of phosphoinositide 3-kinase p85alpha-null platelets characterized by an impaired response to GP VI stimulation. Blood 2003; 102 (02) 541-548
- 16 Zarbock A, Singbartl K, Ley K. Complete reversal of acid-induced acute lung injury by blocking of platelet-neutrophil aggregation. J Clin Invest 2006; 116 (12) 3211-3219
- 17 Rubenfeld GD, Caldwell E, Peabody E. , et al. Incidence and outcomes of acute lung injury. N Engl J Med 2005; 353 (16) 1685-1693
- 18 Brun-Buisson C, Minelli C, Bertolini G. , et al; ALIVE Study Group. Epidemiology and outcome of acute lung injury in European intensive care units. Results from the ALIVE study. Intensive Care Med 2004; 30 (01) 51-61
- 19 Bernard GR, Artigas A, Brigham KL. , et al. The American-European Consensus Conference on ARDS. Definitions, mechanisms, relevant outcomes, and clinical trial coordination. Am J Respir Crit Care Med 1994; 149 (3 Pt 1): 818-824
- 20 Bakowitz M, Bruns B, McCunn M. Acute lung injury and the acute respiratory distress syndrome in the injured patient. Scand J Trauma Resusc Emerg Med 2012; 20: 54
- 21 Kral-Pointner JB, Schrottmaier WC, Horvath V. , et al. Myeloid but not epithelial tissue factor exerts protective anti-inflammatory effects in acid aspiration-induced acute lung injury. J Thromb Haemost 2017; 15 (08) 1625-1639
- 22 Salzmann M, Hoesel B, Haase M. , et al. A novel method for automated assessment of megakaryocyte differentiation and proplatelet formation. Platelets 2018; 29 (04) 357-364
- 23 Veitinger M, Oehler R, Umlauf E. , et al. A platelet protein biochip rapidly detects an Alzheimer's disease-specific phenotype. Acta Neuropathol 2014; 128 (05) 665-677
- 24 Yang H, Wahlmüller FC, Uhrin P. , et al. Proteome analysis of testis from infertile protein C inhibitor-deficient mice reveals novel changes in serpin processing and prostaglandin metabolism. Electrophoresis 2015; 36 (21-22): 2837-2840
- 25 Hirsch E, Bosco O, Tropel P. , et al. Resistance to thromboembolism in PI3Kgamma-deficient mice. FASEB J 2001; 15 (11) 2019-2021
- 26 Matute-Bello G, Frevert CW, Martin TR. Animal models of acute lung injury. Am J Physiol Lung Cell Mol Physiol 2008; 295 (03) L379-L399
- 27 Fruman DA, Ferl GZ, An SS, Donahue AC, Satterthwaite AB, Witte ON. Phosphoinositide 3-kinase and Bruton's tyrosine kinase regulate overlapping sets of genes in B lymphocytes. Proc Natl Acad Sci U S A 2002; 99 (01) 359-364
- 28 Yu J, Zhang Y, McIlroy J, Rordorf-Nikolic T, Orr GA, Backer JM. Regulation of the p85/p110 phosphatidylinositol 3′-kinase: stabilization and inhibition of the p110alpha catalytic subunit by the p85 regulatory subunit. Mol Cell Biol 1998; 18 (03) 1379-1387
- 29 Lagrue AH, Francischetti IM, Guimarães JA, Jandrot-Perrus M. Phosphatidylinositol 3′-kinase and tyrosine-phosphatase activation positively modulate Convulxin-induced platelet activation. Comparison with collagen. FEBS Lett 1999; 448 (01) 95-100
- 30 Gilio K, Munnix IC, Mangin P. , et al. Non-redundant roles of phosphoinositide 3-kinase isoforms alpha and beta in glycoprotein VI-induced platelet signaling and thrombus formation. J Biol Chem 2009; 284 (49) 33750-33762
- 31 Jackson SP, Schoenwaelder SM, Goncalves I. , et al. PI 3-kinase p110beta: a new target for antithrombotic therapy. Nat Med 2005; 11 (05) 507-514
- 32 Moore SF, Smith NR, Blair TA, Durrant TN, Hers I. Critical roles for the phosphatidylinositide 3-kinase isoforms p110β and p110γ in thrombopoietin-mediated priming of platelet function. Sci Rep 2019; 9 (01) 1468
- 33 Canobbio I, Stefanini L, Cipolla L. , et al. Genetic evidence for a predominant role of PI3Kbeta catalytic activity in ITAM- and integrin-mediated signaling in platelets. Blood 2009; 114 (10) 2193-2196
- 34 Rondina MT, Weyrich AS, Zimmerman GA. Platelets as cellular effectors of inflammation in vascular diseases. Circ Res 2013; 112 (11) 1506-1519
- 35 Grommes J, Alard JE, Drechsler M. , et al. Disruption of platelet-derived chemokine heteromers prevents neutrophil extravasation in acute lung injury. Am J Respir Crit Care Med 2012; 185 (06) 628-636
- 36 Duerschmied D, Suidan GL, Demers M. , et al. Platelet serotonin promotes the recruitment of neutrophils to sites of acute inflammation in mice. Blood 2013; 121 (06) 1008-1015
- 37 Petri B, Broermann A, Li H. , et al. von Willebrand factor promotes leukocyte extravasation. Blood 2010; 116 (22) 4712-4719
- 38 Guo Y, Mishra A, Howland E. , et al. Platelet-derived Wnt antagonist Dickkopf-1 is implicated in ICAM-1/VCAM-1-mediated neutrophilic acute lung inflammation. Blood 2015; 126 (19) 2220-2229
- 39 Ortiz-Muñoz G, Mallavia B, Bins A, Headley M, Krummel MF, Looney MR. Aspirin-triggered 15-epi-lipoxin A4 regulates neutrophil-platelet aggregation and attenuates acute lung injury in mice. Blood 2014; 124 (17) 2625-2634
- 40 Looney MR, Nguyen JX, Hu Y, Van Ziffle JA, Lowell CA, Matthay MA. Platelet depletion and aspirin treatment protect mice in a two-event model of transfusion-related acute lung injury. J Clin Invest 2009; 119 (11) 3450-3461
- 41 Schrottmaier WC, Kral JB, Badrnya S, Assinger A. Aspirin and P2Y12 Inhibitors in platelet-mediated activation of neutrophils and monocytes. Thromb Haemost 2015; 114 (03) 478-489
- 42 Nylander S, Kull B, Björkman JA. , et al. Human target validation of phosphoinositide 3-kinase (PI3K)β: effects on platelets and insulin sensitivity, using AZD6482 a novel PI3Kβ inhibitor. J Thromb Haemost 2012; 10 (10) 2127-2136
- 43 Weng Z, Li D, Zhang L. , et al. PTEN regulates collagen-induced platelet activation. Blood 2010; 116 (14) 2579-2581
- 44 Suzuki A, de la Pompa JL, Stambolic V. , et al. High cancer susceptibility and embryonic lethality associated with mutation of the PTEN tumor suppressor gene in mice. Curr Biol 1998; 8 (21) 1169-1178