Thromboxane Synthase Has the Same Pattern of Expression as Platelet Specific Glycoproteins during Human Megakaryocyte Differentiation

Natacha Vitrat; Rémi Letestu; Aline Massé; Vladimir Lazar; William Vainchenker; Najet Debili

doi:10.1055/s-0037-1613905

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2000; 83(05): 759-768
DOI: 10.1055/s-0037-1613905

Review Article

Schattauer GmbH

Thromboxane Synthase Has the Same Pattern of Expression as Platelet Specific Glycoproteins during Human Megakaryocyte Differentiation

Natacha Vitrat

¹From INSERM U 362, Laboratoire associé n°5 du comité de Paris de la Ligue Nationale contre le Cancer, Institut Gustave Roussy, Villejuif, France

,

Rémi Letestu

¹From INSERM U 362, Laboratoire associé n°5 du comité de Paris de la Ligue Nationale contre le Cancer, Institut Gustave Roussy, Villejuif, France

,

Aline Massé

¹From INSERM U 362, Laboratoire associé n°5 du comité de Paris de la Ligue Nationale contre le Cancer, Institut Gustave Roussy, Villejuif, France

,

Vladimir Lazar

¹From INSERM U 362, Laboratoire associé n°5 du comité de Paris de la Ligue Nationale contre le Cancer, Institut Gustave Roussy, Villejuif, France

,

William Vainchenker

¹From INSERM U 362, Laboratoire associé n°5 du comité de Paris de la Ligue Nationale contre le Cancer, Institut Gustave Roussy, Villejuif, France

,

Najet Debili

¹From INSERM U 362, Laboratoire associé n°5 du comité de Paris de la Ligue Nationale contre le Cancer, Institut Gustave Roussy, Villejuif, France

› Institutsangaben

Abstract

Summary

Regulation of the platelet formation process is poorly understood. It has been shown that p45^NF-E2 deficient mice have a profound defect in platelet formation and recently the first platelet/megakaryocytic gene regulated by NF-E2, thromboxane synthase (TXS), has been identified. In this study, we investigated TXS expression as a model of a gene regulated by NF-E2 during MK differentiation. Megakaryocytic cells derived from blood CD34⁺ cells were purified according to their stage of maturation on the basis of expression of CD34, CD41a and CD42a, permitting to define different stages in MK differentiation. By means of real-time quantitative RT-PCR, we could determine that the level of TXS increased during differentiation in parallel with the expression of c-mpl and GPIIb (CD41). However, amounts of TXS transcripts increased about 1.6-fold more than that of GPIIb or c-mpl transcripts during maturation. Expression of TXS and MK specific proteins such as CD41a, CD42a and vWF was also correlated in maturing MKs. In addition, staining by anti-TXS antibody of proplatelet bearing MKs was not increased in comparison to that observed in mature MK, suggesting that TXS is not upregulated during platelet formation. In addition, we investigated whether TXS and cyclooxygenase could be involved in platelet formation by adding aspirin into the cultures. No significant decrease of platelet production was observed.

In conclusion, this study shows that TXS is coordinately expressed with the other platelet proteins during MK differentiation but is not directly involved in platelet formation.

Keywords

Megakaryocyte - TXS

Volltext

Referenzen

References
1 Bartley TD, Bogenberger J, Hunt P, Li YS, Lu HS, Martin F, Chang MS, Samal B, Nichol JL, Swift S, Johnson MJ, Hsu RY, Parker VP, Suggs S, Skrine JD, Merewether LA, Clogston C, Hsu E, Hokom MM, Hornkohl A, Choi E, Pangelinan M, Sun Y, Mar V, McNinch J, Simonet L, Jacobsen F, Xie C, Shutter J, Chute H, Basu R, Selander L, Trollinger D, Sieu L, Padilla D, Trail G, Elliot G, Izumi R, Covey T, Crouse J, Garcia A, Xu W, Del Castillo J, Biron J, Cole S, Hu CT, Pacifici R, Ponting I, Saris C, Wen D, Yung YP, Lin H, Bosselman RA. Identification and cloning of a megakaryocyte growth and development factor that is a ligand for the cytokine receptor Mpl. Cell 1994; 77: 1117-24.
2 de Sauvage FJ, Hass PE, Spencer SD, Malloy BE, Gurney AL, Spencer SA, Darbonne WC, Henzel WJ, Wong SC, Kuang W-J, Oles KJ, Hultgren B, Solberg Jr LA, Goeddel DV, Eaton DL. Stimulation of megakaryocytopoiesis and thrombopoiesis by the c-Mpl ligand. Nature 1994; 369: 533-8.
3 Lok S, Kaushansky K, Holly RD, Kuijper JL, Lofton-Day CE, Oort PJ, Grant FJ, Heipel MD, Burhead SK, Kramer JM, Bell LA, Sprecher CA, Blumberg H, Johnson R, Prunkard D, Ching AFT, Bailey MC, Forstrom JW, Buddle MM, Osborn SG, Evans SJ, Sheppard PO, Presnell SR, O’Hara PJ, Hagen FS, Roth GR, Foster DC. Murine thrombopoietin: expression cloning, cDNA sequence and stimulation of platelet production in vivo. Nature 1994; 369: 565-8.
4 Kaushansky K. Thrombopoietin, the primary regulator of platelet production. Blood 1995; 86: 419-31.
5 Gewirtz AM, Poncz M. Megakaryocytopoiesis and platelet production. In: Hematology. Basic principales and practice. Hoffman R, Benz EJ, Schattil SJ. eds. New York: Churchill Livingstone; 1991: 1148-57.
6 Hoffman R. Regulation of megakaryocytopoiesis. Blood 1989; 74: 1196-212.
7 Vainchenker W, Kieffer N, Cramer E, Vinci G, Debili N, Henri A, Titeux M, Guichard J, Mishal Z, Breton-Gorius J. Expression of platelet proteins during maturation of human megakaryocytes and of the K562/HEL cell lines. In: Megakaryocyte development and function. Levine RF, Williams N, Levin J, Evatt BL. eds. New York: Liss Alan; 1986: 301-17.
8 Briddell RA, Brandt JE, Straneva JE, Srour EF, Hoffman R. Characterization of the human Burst-Forming Unit-Megakaryocyte. Blood 1989; 74: 145-51.
9 Young JC, Bruno E, Luens KM, Wu S, Backer M, Murray LJ. Thrombopoietin stimulates megakaryocytopoiesis, myelopoiesis and expansion of CD34⁺ progenitor cells from single CD34⁺Thy⁻1⁺Lin⁻ primitive progenitor cells. Blood 1996; 88: 1619-31.
10 Berardi AC, Wang A, Levine JD, Lopez P, Scaden DT. Functional isolation and characterization of human hematopoietic stem cells. Science 1995; 267: 104-8.
11 Alexander WS, Roberts AW, Nicola NA, Li R, 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.
12 de Sauvage FJ, Carver-Moore K, Luoh S-M, Ryan A, Dowd M, Eaton DL, Moore MW. Physiological regulation of early and late stages of megakaryocytopoiesis by thrombopoietin. J Exp Med 1996; 183: 651-6.
13 Kimura S, Roberts AW, Metcalf D, Alexander WS. Hematopoietic stem cell deficiencies in mice lacking c-Mpl, the receptor for thrombopoietin. Proc Natl Acad Sci USA 1998; 95: 1195-200.
14 Murone M, Carpenter DA, de Sauvage FJ. Hematopoietic deficiencies in c-mpl and TPO knockout mice. Stem cells 1998; 16: 1-6.
15 Debili N, Issaad C, Masse JM, Guichard J, Katz A, Breton-Gorius J, Vainchenker W. Expression of CD34 and platelet glycoproteins during human megakaryocytic differentiation. Blood 1992; 80: 3022-35.
16 Debili N, Wendling F, Katz A, Guichard J, Breton-Gorius J, Hunt P, Vainchenker W. The Mpl-Ligand or thrombopoietin or megakaryocyte growth and differentiative factor has both direct proliferative and differentiative activities on human megakaryocyte progenitors. Blood 1995; 86: 2516-25.
17 Berridge MV, Ralph SJ, Tan AS. Cell lineage antigens of the stem cellmegakaryocyte platelet lineage are associated with the platelet IIb-IIIa glycoprotein complex. Blood 1985; 66: 76-85.
18 Tronik-Le Roux D, Roullot V, Schweitzer A, Berthier R, Marguerie G. Suppression of erythro-megakaryocytopoiesis and the induction of reversible thrombocytopenia in mice transgenic for the thymidine kinase gene targeted by the platelet glycoprotein aIIb promoter. J Exp Med 1995; 181: 2141-51.
19 Troppel PVR, Vernet M, Poujol C, Pointu H, Nurden P, Marguerie G, Tronik-le Roux D. A 2.7-kb portion of the 5’ flanking region of the murine glycoprotein alphaIIb gene is transcriptionally active in primitive hematopoietic cells. Blood 1997; 90: 2995-3004.
20 Rabellino EM, Nachman RL, Williams N, Winchester R, Ross GD. Human megakaryocytes. I Characterization of the membrane and cytoplasmic components of isolated marrow megakaryocytes. J Exp Med 1979; 149: 1273-87.
21 Rabellino EM, Levene RB, Leung LLK, Nachman RL. Human megakaryocytes. II Expression of platelet proteins in early marrow megakaryocytes. J Exp Med 1981; 154: 85-100.
22 Vinci G, Tabilio A, Deschamps J-F, Van Haeke D, Henri A, Guichard J, Tetteroo P, Lansdorp PM, Hercend T, Vainchenker W, Breton-Gorius J. Immunological study of in vitro maturation of human megakaryocytes. Br J Haematol 1984; 56: 589-605.
23 Block KL, Ravid K, Phung QH, Poncz M. Characterization of regulatory elements in the 5’-flanking region of the rat GpIIb gene by studies in a primary rat bone marrow culture system. Blood 1994; 84: 3385-93.
24 Deveaux S, Filippe A, Lemarchandel V, Ghysdael J, Roméo P-H, Mignotte V. Analysis of the thrombopoietin receptor (MPL) promoter implicates GATA and Ets proteins in the coregulation of megakaryocyte-specific genes. Blood 1996; 87: 4678-85.
25 Hashimoto Y, Ware J. Identification of essential GATA and Ets binding motifs within the promoter of the platelet glycoprotein Ib a gene. J Biol Chem 1995; 270: 24532-9.
26 Hickey MJ, Roth GJ. Characterization of the gene encoding human platelet glycoprotein IX. J Biol Chem 1993; 268: 3438-43.
27 Lanza F, Morales M, de la Salle C, Cazenave J-P, Clemetson KJ, Shimomura T, Phillips DR. Cloning and characterization of the gene encoding the human platelet glycoprotein V. J Biol Chem 1993; 268: 20801-7.
28 Lemarchandel V, Ghysdael J, Mignotte V, Rahuel C, Roméo PH. Gata and Ets cis-acting sequence mediate megakaryocyte-specific expression. Mol Cell Biol 1993; 13: 668-76.
29 Pan J, McEver RP. Characterization of the promoter for the human P-selectin gene. J Biol Chem 1993; 268: 22600-8.
30 Prandini MH, Uzan G, Martin F, Thevenon D, Marguerie G. Characterization of a specific erythromegakaryocytic enhancer within the glycoprotein IIb promoter. J Biol Chem 1992; 267: 10370-4.
31 Ravid K, Doi T, Beeler L, Kuter DJ, Rosenberg RD. Transcriptional regulation of the rat platelet factor 4 gene: interaction between an enhancer/ silencer domain and the GATA-1 site. Mol Cell Biol 1991; 11: 6116-27.
32 Andrews NC, Erjument-Bromage H, Davidson MB, Tempst P, Orkin SH. Erythroid transcription factor NFE-2 is a haematopoietic-specific basic-leucine zipper protein. Nature 1993; 362: 722-8.
33 Mignotte V, Eleouet JF, Raich N, Roméo P-H. Cis- and transacting elements invoved in the regulation of the erythroid promoter of the human porphobilinogen deaminase gene. Proc Natl Acad Sci USA 1989; 86: 6548-52.
34 Shivdasani RA, Rosenblatt MF, Zucker-Franklin D, Jackson CW, Hunt P, Saris CJM, Orkin SH. Transcription factor NF-E2 is required for platelet formation independent of actions of thrombopoietin/MGDF in megakaryocyte development. Cell 1995; 81: 695-704.
35 Shivdasani RA, Fielder P, Keller GA, Orkin SH, de Sauvage FJ. Regulation of the serum concentration of thrombopoietin in thrombocytopenic NF-E2 knockout mice. Blood 1997; 90: 1821-7.
36 Chan JY, Kwong M, Lu R, Chang J, Wang B, Yen TSB, Kan YW. Targeted disruption of the ubiquitous CNC-bZIP transcription factor, Nrf-1, results in anemia and embryonic lethality in mice. EMBO J 1998; 17: 1779-87.
37 Martin F, van Deursen JM, Shivdasani RA, Jackson CW, Troutman AG, Ney PA. Erythroid maturation and globin gene expression in mice with combined deficiency in NF-E2 and nrf-2. Blood 1998; 91: 3459-66.
38 Deveaux S, Cohen-Kaminsky S, Shivdasani RA, Andrews NC, Filipe A, Kuzniak I, Orkin SH, Roméo PH, Mignotte V. p45 NF-E2 regulates expression of thromboxane synthase in megakaryocytes. EMBO J 1997; 16: 5654-61.
39 Lee KD, Baek SJ, Shen RF. Multiple factors regulating the expression of human thromboxane synthase gene. Biochem J 1996; 319: 783-91.
40 Zhang L, Xiao H, Schultz RA, Shen R-F. Genomic organization, chromosomal localization, and expression of the murine thromboxane synthase gene (Tbxas1). Genomics 1997; 45: 519-28.
41 Zhang L, Chase B, Shen RF. Molecular cloning and expression of murine thromboxane synthase. Biochem Biophys Res Commun 1993; 194: 741-8.
42 Nüsing R, Wernet MP, Ullrich V. Production and characterization of polyclonal and monoclonal antibodies against human thromboxane synthase. Blood 1990; 76: 80-5.
43 Norol F, Vitrat N, Cramer E, Guichard J, Burstein SA, Vainchenker W, Debili N. Effects of cytokines on platelet production from blood and marrow CD34 positive cells. Blood 1998; 91: 830-43.
44 Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-9.
45 Haurand M, Ullrich V. Isolation and characterization of thromboxane synthase from human platelets as a cytochrome P-450. J Biol Chem 1985; 260: 15059-67.
46 Tone Y, Miyata A, Hara S, Yukawa S, Tanabe T. Abundant expression of thromboxane synthase in rat macrophages. FEBS lett 1994; 340: 241-4.
47 Worthington RE, Nakeff A. Thromboxane synthesis in megakaryocytes isolated by centrifugal elutriation. Blood 1981; 58: 175-8.
48 Ihara H, Yokoyama C, Miyata A, Kosaka T, Nusing R, Ullrich V, Tanabe T. Induction of thromboxane synthase and prostaglandin endoperoxide synthase mRNAs in huma erythroleukemia cells by phorbol ester. FEBS lett 1992; 306: 161-4.
49 Matijevic-Aleksic N, Sanduja SK, Wang LH, Wu KK. Differential expression of thromboxane A synthase and prostaglandin H synthase in megakaryocytic cell line. Biochim Biophys Acta 1995; 1269: 167-75.
50 Long MW, Heffner CH, Williams JL, Peters C, Prochownik EV. Regulation of megakaryocyte phenotype in human erythroleukemia cells. J Clin Invest 1990; 85: 1072-84.
51 Tabilio AJ, Rosa P, Testa U, Kieffer N, Nurden AT, Delcanizo MC, Breton-Gorius J, Vainchenker W. Expression of platelet membrane glycoproteins and alpha granule proteins by human erythroleukemia cell line (HEL). EMBO J 1984; 03: 453-9.
52 Toki T, Itoh J, Arai K, Kitazawa J, Yokoyama M, Igarashi K, Yamamoto M, Ito E. Abundant expression of erythroid transcription factor p45 NF-E2 mRNA in human peripheral granulocytes. Biochem Biophys Res Commun 1996; 219: 760-5.
53 Mohler J, Vani K, Leung S, Epstein A. Segmentally restricted, cephalic expression of a leucine zipper gene during Drosophilia embryogenesis. Mech Dev 1997; 34: 3.
54 Andrews NC, Kotkow KJ, Ney PA, Erdjument-Bromage H, Tempst P, Orkin SH. The ubiquitous subunit of erythroid transcription factor NF-E2 is a small basic-leucine zipper protein related to the v-maf oncogene. Proc Natl Acad Sci USA 1993; 90: 11488-92.
55 Igarashi K, Kataoka K, Itoh K, Hayashi N, Nishizawa M, Yamamoto M. Regulation of transcription by dimerization of erythroid factor NF-E2 p45 with small Maf proteins. Nature 1994; 367: 568-72.
56 Kataoka K, Igarashi K, Itoh K, Fujiwara KT, Noda M, Yamamoto M, Nishizawa M. Small maf proteins heterodimerize with Fos and may act as competitive repressors of the NF-E2 transcription factor. Mol Cell Biol 1995; 15: 2180-90.
57 Lecine P, Blank V, Shivadasani R. Characterization of the hematopoietic transcription factor NF-E2 in primary murine megakaryocytes. J Biol Chem 1998; 273: 7572-8.
58 Shavit JA, Motohashi H, Onodera K, Akasaka J, Yamamoto M, Engel JD. Impaired megakaryopoiesis and behavioral defects in mafG-null mutant mice. Genes dev 1998; 12: 2164-74.
59 Chan JY, Han X-L, Kan Y-W. Isolation of cDNA encoding the human NF-E2 protein. Proc Natl Acad Sci USA 1993; 90: 11366-70.
60 Caterina JJ, Donze D, Sun CW, Ciavatta DJ, Townes TM. Cloning and functional characterization of LCR-F1: a bZIP transcription factor that activates erythroid-specific, human globin gene expression. Nucleic Acids Res 1994; 22: 2383-91.
61 Moi P, Chan K, Cao A, Kan YW. Isolation of NF-E2 related factor 2 (Nrf2), a NF-E2-like basic leucine zipper transcriptional activator that binds to the tandem NF-E2/AP1 repeat of the β-globin locus control region. Proc Natl Acad Sci USA 1994; 91: 9926-30.
62 Itoh K, Igarashi K, Hayashi N, Nishizawa M, Yamamoto M. Cloning and characterization of a novel erythroid cell-derived CNC family transcriptipn factor heterodimerizing with the small Maf family proteins. Mol Cell Biol 1995; 15: 4184-93.
63 Maclouf J, Folco G, Patrono C. Eicosanoids and iso-eicosanoids: constitutive, inducible and transcellular biosynthesis in vascular disease. Thromb Haemost 1998; 1998: 691-705.
64 Mestel F, Oetliker O, Beck E, Felix R, Imbach P, Wagner HP. Severe bleeding associated with defective thromboxane synthase. Lancet 1980; i: 157.
65 Rao AK, Koike K, Day HJ, Smith JB, Holmsen H. Bleeding disorder associated with albumin-dependent partial deficiency in platelet thromboxane production. Effect of albumin on arachidonate metabolism in platelets. Am J Clin Pathol 1985; 83: 687-96.
66 Sinzinger H, Reiter S, Peskar BA. Deficiencies of the prostaglandin system: III. A partial thromboxane synthase defect. Wien Klin Wochenschr 1985; 97: 561-5.
67 Weiss HJ, Lages BA. Possible congenital defect in platelet thromboxane synthase. Lancet 1977; i: 760-1.
68 Wu KK, Minkoff IM, Rossi EC, Chen YC. Hereditary bleeding disorder due to a primary defect in platelet release reaction. Br J Hematol 1981; 47: 241-9.
69 Tsujii M, Kawano S, Tsuji S, Sawaoka H, Hori M, DuBois RN. Cyclooxygenase regulates angiogenesis induced by colon cancer cells. Cell 1998; 93: 705-16.
70 Bikfavi A, Han ZC, Fuhrmann G. Interaction of fibroblast growth factor (FGF) with megakaryocytopoiesis and demonstration of FGF receptor expression in megakaryocytes and megakaryocyte-like cells. Blood 1992; 80: 1905-13.
71 Bobik R, Hong Y, Breier G, Martin JF, Erusalimski JD. Thombopoietin stimulates VEGF from c-mpl-expressing cell lines and haematopoietic progenitors. FEBS Lett 1998; 423: 10-4.
72 Möhle R, Green D, Moore MAS, Nachman RL, Rafil S. Constituve production and thrombin-induced release of vascular endothelial growth factor by human megakaryocytes and platelets. Proc Natl Acad Sci USA 1997; 94: 663-8.
73 Roy A, Fava T, Casey J, Ron W, Harold L, Lilian B. Synthesis of transforming growth factor-β1 by megakaryocytes and its localization to megakaryocyte and platelet α-granules. Blood 1990; 76: 1946-55.
74 Mitjavila MT, Vinci G, Villeval JL, Kieffer N, Henri A, Testa U, Breton-Gorius J, Vainchenker W. Human platelet alpha granules contain a non specific inhibitor of megakaryocyte colony formation: its relationship to type β transforming growth factor (TGF-β). J Cell Physiol 1988; 134: 93-100.