Thrombopoietin Increases Platelet Sensitivity to α-Thrombin via Activation of the ERK2-cPLA2 Pathway

Gijsbert van Willigen; Gertie Gorter; Jan-Willem N. Akkerman

doi:10.1055/s-0037-1613872

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2000; 83(04): 610-616
DOI: 10.1055/s-0037-1613872

Commentary

Schattauer GmbH

Thrombopoietin Increases Platelet Sensitivity to α-Thrombin via Activation of the ERK2-cPLA₂ Pathway

Gijsbert van Willigen

¹From the Department of Haematology, University Hospital Utrecht, and Institute for Biomembranes, University of Utrecht, the Netherlands

,

Gertie Gorter

¹From the Department of Haematology, University Hospital Utrecht, and Institute for Biomembranes, University of Utrecht, the Netherlands

,

Jan-Willem N. Akkerman

¹From the Department of Haematology, University Hospital Utrecht, and Institute for Biomembranes, University of Utrecht, the Netherlands

› Author Affiliations

Abstract

Summary

Thrombopoietin (TPO) regulates stem cell proliferation and maturation of megakaryocytes by activating the c-Mpl-receptor, a member of the hematopoietic cytokine family. As human platelets possess c-Mplreceptors and supraphysiological concentrations of TPO trigger platelet aggregation and secretion, we searched for the signalling pathways through which the c-Mpl-receptor might activate platelets. A physiological concentration of TPO (20 ng/mL) did not trigger platelet functions, but increased their sensitivity to α-thrombin resulting in a 4-fold faster dense granule secretion. The effect of TPO was abolished by indomethacin and caused by synergism with signal generation by α-thrombin at the level of the cytosolic phospholipase A₂ (cPLA₂) pathway resulting in more arachidonate release, cPLA₂ phosphorylation and thromboxane A₂ formation. A similar synergism was seen at the level of extracellular signal-regulated kinase 2 (ERK2 or p42-MAPK). These data suggest, that TPO increases the sensitivity of platelets to α-thrombin by enhancing cPLA₂ activation via the ERK2-cPLA₂ pathway.

Keywords

Thrombopoietin - platelet sensitisation - cytosolic phospholipase A₂ - thromboxane A₂ - extracellular signal-regulated kinase

Full Text

References

References
1 Alexander WS, Roberts AW, Nicola NA, Li RL, Metcalf D. Deficiencies in progenitor cells of multiple hematopoietic lineages and defective megakaryocytopoiesis in mice lacking the thrombopoietin receptor c-mpl. Blood 1996; 87: 2162-70.
2 Carver-Moore K, Broxmeyer HE, Luoh SM. et al. Low levels of erythroid and myeloid progenitors in thrombopoietin- and c-mpl-deficient mice. Blood 1996; 88: 803-8.
3 De Sauvage F, Carver-Moore K, Luoh S. et al. Physiological regulation of early and late stages of megakaryocytopoiesis by thrombopoietin. J Exp Med 1996; 183: 651-6.
4 De Sauvage FJ, Hass PE, Spencer SD. et al. Stimulation of megakaryocytopoiesis and thrombopoiesis by the c-Mpl ligand. Nature 1994; 369: 533-8.
5 Lok S, Kaushansky K, Holly RD. et al. Cloning and expression of murine thrombopoietin cDNA and stimulation of platelet production in vivo . Nature 1994; 369: 565-8.
6 Bartley TD, Bogenberger J, Hunt P. et al. Identification and cloning of a megakaryocyte growth and development factor that is a ligand for the cytokine receptor Mpl. Cell 1994; 77: 1117-24.
7 Wendling F, Varlet P, Charon M, Tambourin P. MPLV: A retrovirus complex inducing an acute myeloproliferative disorder in adult mice. Virology 1986; 149: 242-6.
8 Souyri M, Vignon I, Pencioelli J-F, Heard J-M, Tambourin P, Wendling F. A putative truncated cytokine receptor gene transduced by the myeloproliferative leukemia virus immortalizes hematopoietic progenitors. Cell 1990; 63: 1137-47.
9 Debili N, Wendling F, Cosman D. et al. The Mpl receptor is expressed in the megakaryocytic lineage from late progenitors to platelets. Blood 1995; 85: 391-401.
10 Broudy VC, Lin NL, Sabath DF. Human platelets display high-affinity receptors for thrombopoietin. Blood 1997; 89: 1896-904.
11 Gurney AL, Carver-Moore K, De Sauvage FJ, Moore MW. Thrombocytopenia in c-mpl-deficient mice. Science 1994; 265: 1445-7.
12 Muraoka K, Ishii E, Tsuji K. et al. Defective response to thrombopoietin and impaired expression of c-mpl mRNA of bone marrow cells in congenital amegakaryocytic thrombocytopenia. Br J Haematol 1997; 96: 287-92.
13 Ballmaier M, Schulze H, Strauss G. et al. Thrombopoietin in patients with congenital thrombocytopenia and absent radii: Elevated serum levels, normal receptor expression, but defective reactivity to thrombopoietin. Blood 1997; 90: 612-9.
14 Drachman JG, Sabath DF, Fox NE, Kaushansky K. Thrombopoietin signal transduction in purified murine megakaryocytes. Blood 1997; 89: 483-92.
15 Tortolani PJ, Johnston JA, Bacon CM. et al. Thrombopoietin induces tyrosine phosphorylation and activation of the Janus kinase, JAK2. Blood 1995; 85: 3444-51.
16 Pallard C, Gouilleux F, Bénit L. et al. Thrombopoietin activates a STAT5- like factor in hematopoietic cells. EMBO J 1995; 14: 2847-56.
17 Yamada M, Komatsu N, Okada K, Kato T, Miyazaki H, Miura Y. Thrombopoietin induces tyrosine phosphorylation and activation of mitogen-activated protein kinases in a human thrombopoietin- dependent cell line. Biochem Biophys Res Commun 1995; 217: 230-7.
18 Bacon CM, Tortolani PJ, Shimosaka A, Rees RC, Longo DL, O’Shea JJ. Thrombopoietin (TPO) induces tyrosine phosphorylation and activation of STAT5 and STAT3. FEBS Lett 1995; 370: 63-8.
19 Nagata Y, Todokoro K. Thrombopoietin induces activation of at least two distinct signaling pathways. FEBS Lett 1995; 377: 497-501.
20 Rodríguez-Liñares B, Watson SP. Thrombopoietin potentiates activation of human platelets in association with JAK2 and TYK2 phosphorylation. Biochem J 1996; 316: 93-8.
21 Miyakawa Y, Oda A, Druker BJ. et al. Thrombopoietin induces tyrosine phosphorylation of Stat3 and Stat5 in human blood platelets. Blood 1996; 87: 439-46.
22 Montrucchio G, Brizzi MF, Calosso G, Marengo S, Pegoraro L, Camussi G. Effects of recombinant human megakaryocyte growth and development factor on platelet activation. Blood 1996; 87: 2762-8.
23 Miyakawa Y, Oda A, Druker BJ. et al. Recombinant thrombopoietin induces rapid protein tyrosine phosphorylation of Janus kinase 2 and Shc in human blood platelets. Blood 1995; 86: 23-7.
24 Kojima H, Hamazaki Y, Nagata Y, Todokoro K, Nagasawa T, Abe T. Modulation of platelet activation in vitro by thrombopoietin. Thromb Haemost 1995; 74: 1541-5.
25 Oda A, Miyakawa Y, Druker BJ. et al. Thrombopoietin primes human platelet aggregation induced by shear stress and by multiple agonists. Blood 1996; 87: 4664-70.
26 Van Willigen G, Akkerman J-WN. Protein kinase C and cyclic AMP regulate reversible exposure of binding sites for fibrinogen on the glycoprotein IIB-IIIA complex of human platelets. Biochem J 1991; 273: 115-20.
27 Akkerman J-WN, Holmsen H. Interrelationships among platelet responses: Studies on the burst in proton liberation, lactate production, and oxygen uptake during platelet aggregation and Ca²⁺ secretion. Blood 1981; 57: 956-66.
28 Bligh EG, Dyer WJ. A rapid method of total lipid extraction and purification. Can J Physiol 1959; 37: 911-8.
29 Bekkers ACAPA, Van der Vuurst H, Van Willigen G, Akkerman JWN, Verheij HM. Targeting of porcine pancreatic phospholipase A₂ to human platelets: Introduction of an RGD sequence by genetic engineering. Thromb Haemost 1995; 74: 1138-44.
30 De Vries-Smits AMM, Burgering BMT, Leevers SJ, Marshall CJ, Bos JL. Involvement of p21ras in activation of extracellular signal-regulated kinase 2. Nature 1992; 357: 602-604.
31 Kramer RM, Roberts EF, Manetta JV, Hyslop PA, Jakubowski JA. Thrombin-induced phosphorylation and activation of Ca²⁺-sensitive cytosolic phospholipase A₂ in human platelets. J Biol Chem 1993; 268: 26796-804.
32 Gordon RD, Leighton IA, Campbell DG. et al. Cloning and expression of cytosolic phospholipase A₂ (cPLA₂) and a naturally occurring variant – Phosphorylation of Ser⁵⁰⁵ of recombinant cPLA₂ by p42 mitogen-activated protein kinase results in an increase in specific activity. Eur J Biochem 1996; 238: 690-7.
33 Leslie CC. Properties and regulation of cytosolic phospholipase A₂ . J Biol Chem 1997; 272: 16709-12.
34 Borsch-Haubold AG, Pasquet S, Watson SP. Direct inhibition of cyclooxygenase-1 and -2 by the kinase inhibitors SB 203580 and PD 98059. SB 203580 also inhibits thromboxane synthase. J Biol Chem 1998; 273: 28766-72.
35 Kramer RM, Sharp JD. Structure, function and regulation of Ca²⁺-sensitive cytosolic phospholipase A₂ (cPLA₂). FEBS Lett 1997; 410: 49-53.
36 Böhm SK, Kong WY, Brömme D. et al. Molecular cloning, expression and potential functions of the human proteinase-activated receptor-2. Biochem J 1996; 314: 1009-16.
37 Ishihara H, Connolly AJ, Zeng DW. et al. Protease-activated receptor 3 is a second thrombin receptor in humans. Nature 1997; 386: 502-6.
38 Coughlin SR. Protease-activated receptors start a family. Proc Natl Acad Sci USA 1994; 91: 9200-2.
39 Gerrard JM, McNicol A, Saxena SP. Protein kinase C, membrane fusion and platelet granule secretion. Biochem Soc Trans 1993; 21: 289-93.
40 Shock DD, He K, Wencel-Drake JD, Parise LV. Ras activation in platelets after stimulation of the thrombin receptor, thromboxane A₂ receptor or protein kinase C. Biochem J 1997; 321: 525-30.
41 Kramer RM, Roberts EF, Um SL. et al. p38 Mitogen-activated protein kinase phosphorylates cytosolic phospholipase A₂ (cPLA₂) in thrombin-stimulated platelets – Evidence that proline-directed phosphorylation is not required for mobilization of arachidonic acid by cPLA₂ . J Biol Chem 1996; 271: 27723-9.
42 Börsch-Haubold AG, Bartoli F, Asselin J. et al. Identification of the phosphorylation sites of cytosolic phospholipase A₂ in agonist-stimulated human platelets and HeLa cells. J Biol Chem 1998; 273: 4449-58.
43 de Carvalho M, McCormack AL, Olson E. et al. Identification of phosphorylation sites of human 85-kDa cytosolic phospholipase A₂ expressed in insect cells and present in human monocytes. J Biol Chem 1996; 271: 6987-97.
44 Papkoff J, Chen R-H, Blenis J, Forsman J. p42 Mitogen-activated protein kinase and p90 ribosomal S6 kinase are selectively phosphorylated and activated during thrombin- induced platelet activation and aggregation. Mol Cell Biol 1994; 14: 463-72.
45 Nakashima S, Chatani Y, Nakamura M, Miyoshi N, Kohno M, Nozawa Y. Tyrosine phosphorylation and activation of mitogen-activated protein kinases by thrombin in human platelets: Possible involvement in late arachidonic acid release. Biochem Biophys Res Commun 1994; 198: 497-503.
46 Börsch-Haubold AG, Kramer RM, Watson SP. Cytosolic phospholipase A₂ is phosphorylated in collagen- and thrombin-stimulated human platelets independent of protein kinase C and mitogen-activated protein kinase. J Biol Chem 1995; 270: 25885-92.
47 Wells JA, De Vos AM. Hematopoietic receptor complexes. Annu Rev Biochem 1996; 65: 609-34.
48 Alexander WS, Maurer AB, Novak U, Harrison-Smith M. Tyrosine-599 of the c-Mpl receptor is required for Shc phosphorylation and the induction of cellular differentiation. EMBO J 1996; 15: 6531-40.
49 Morita H, Tahara T, Matsumoto A, Kato T, Miyazaki H, Ohashi H. Functional analysis of the cytoplasmic domain of the human Mpl receptor for tyrosine-phosphorylation of the signaling molecules, proliferation and differentiation. FEBS Lett 1996; 395: 228-34.
50 Drachman JG, Kaushansky K. Dissecting the thrombopoietin receptor: Functional elements of the Mpl cytoplasmic domain. Proc Natl Acad Sci USA 1997; 94: 2350-5.
51 Gurney AL, Wong SC, Henzel WJ, De Sauvage FJ. Distinct regions of c-Mpl cytoplasmic domain are coupled to the JAK-STAT signal transduction pathway and Shc phosphorylation. Proc Natl Acad Sci USA 1995; 92: 5292-6.
52 O’Malley CJ, Rasko JEJ, Basser RL. et al. Administration of pegylated recombinant human megakaryocyte growth and development factor to humans stimulates the production of functional platelets that show no evidence of in vivo activation. Blood 1996; 88: 3288-98.
53 Kuter DJ, Rosenberg RD. Appearance of a megakaryocyte growth-promoting activity, megapoietin, during acute thrombocytopenia in the rabbit. Blood 1994; 84: 1464-72.
54 Peng JP, Friese P, Wolf RF. et al. Relative reactivity of platelets from thrombopoietin- and interleukin-6 treated dogs. Blood 1996; 87: 4158-63.
55 Farese AM, Hunt P, Boone T, MacVitte TJ. Recombinant human megakaryocyte growth and development factor stimulates thrombocytopoiesis in normal nonhuman primates. Blood 1996; 86: 54-59.
56 Harker LA, Hunt P, Marzec UM. et al. Regulation of platelet production and function by megakaryocyte growth and development factor in nonhuman primates. Blood 1996; 87: 1833-44.
57 Harker LA, Marzec UM, Hunt P. et al. Dose-response effects of pegylated human megakaryocyte growth and development factor on platelet production and function in nonhuman primates. Blood 1996; 88: 511-21.
58 Basser RL, Rasko JE, Clarke K. et al. Randomized blinded, placebo-controlled phase I trial of pegylated recombinant human megakaryocyte growth and development factor with filgrastim after dose-intensive chemotherapy in patient with advanced cancer. Blood 1997; 89: 3118-28.