Regulation of PAI-1 Concentration in Platelets by Systemic Administration of Antisense Oligonucleotides to Rats

Zofia Pawlowska; Ewa Chabielska; Anna Kobylanska; Anna Maciaszek; Maria Swiatkowska; Wlodzimierz Buczko; Wojciech J. Stec; Czeslaw S. Cierniewski

doi:10.1055/s-0037-1615968

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2001; 85(06): 1086-1089
DOI: 10.1055/s-0037-1615968

Review Article

Schattauer GmbH

Regulation of PAI-1 Concentration in Platelets by Systemic Administration of Antisense Oligonucleotides to Rats

Zofia Pawlowska

¹Department of Biophysics, Medical University in Lodz, Lodz, Poland

,

Ewa Chabielska

²Department of Pharmacodynamics, Medical University in Bialystok, Lodz, Poland

,

Anna Kobylanska

³Department of Bioorganic Chemistry of the Center for Molecular and Macromolecular Studies, Polish Academy of Sciences, Lodz, Poland

,

Anna Maciaszek

³Department of Bioorganic Chemistry of the Center for Molecular and Macromolecular Studies, Polish Academy of Sciences, Lodz, Poland

,

Maria Swiatkowska

¹Department of Biophysics, Medical University in Lodz, Lodz, Poland

,

Wlodzimierz Buczko

²Department of Pharmacodynamics, Medical University in Bialystok, Lodz, Poland

,

Wojciech J. Stec

³Department of Bioorganic Chemistry of the Center for Molecular and Macromolecular Studies, Polish Academy of Sciences, Lodz, Poland

,

Czeslaw S. Cierniewski

¹Department of Biophysics, Medical University in Lodz, Lodz, Poland

› Institutsangaben

Abstract

Summary

In this report we tested the effect of oligodeoxyribonucleotides antisense to PAI-1 mRNA administered into rats on PAI-1 concentration in platelets. Low doses of the antisense oligonucleotide (MPO-16R) reduced PAI-1 activity, both in rat blood plasma and platelet lysates by 20.5% and 28.7%, respectively. There was no change in platelet count after treatment with MPO-16R but treated platelets showed lower aggregability as compared with controls (37 ± 13% and 54 ± 12%, respectively). In an experimental model of rat arterial thrombosis, low doses of MPO-16R caused a significant delay in the occlusion time (31.8%). These data further support for the role of PAI-1 as a major determinant of arterial thrombolysis resistance and for the first time demonstrate the possibility of reduction of platelet PAI-1 concentration by antisense approach.

Key words

PAI-1 - antisense oligonucleotides - blood platelets

Volltext

Referenzen

References
1 Erickson LA, Ginsberg MH, Loskutoff DJ. Detection and partial characterization of an inhibitor of plasminogen activator in human platelets. J Clin Invest 1984; 74: 1465-72.
2 Kruithof EK, Tran Thang C, Bachmann F. Studies on the release of a plasminogen activator inhibitor by human platelets. Thromb Haemost 1986; 55: 201-5.
3 Kunitada S, FitzGerald GA, FitzGerald DJ. Inhibition of clot lysis and decreased binding of tissue-type plasminogen activator as a consequence of clot retraction. Blood 1992; 79: 1420-27.
4 Bajzar L, Tracy PB. Platelet factor Va is responsible for the majority of the antifibrinolytic effect of platelets. Blood 1995; 86: 74a (abstract).
5 Sprengers ED, Akkerman JW, Jansen BG. Blood platelet plasminogen activator inhibitor: two different pools of endothelial cell type plasminogen activator inhibitor in human blood. Thromb Haemost 1986; 55: 325-9.
6 Kruithof EKO, Nicolosa G, Bachmann F. Plasminogen activator inhibitor 1: development of a radioimmunoassay and observations on its plasma concentration during venous occlusion and after platelet aggregation. Blood 1987; 70: 1645-53.
7 Urden G, Chmielewska J, Carlsson T, Wiman B. Immunological relationship between plasminogen activator inhibitors from different sources. Thromb Haemost 1987; 57: 29-34.
8 Robbie LA, Young SP, Bennett B, Booth NA. Thrombi formed in a Chandler loop mimic human arterial thrombi in structure and PAI-1 content and distribution. Thromb Haemost 1997; 77: 510-15.
9 Hantgan RR, Jerome WG, Handt S. Platelets and endothelial cells act in concert to delay thrombolysis – evidence from an in vitro model of the human occlusive thrombus. Thromb Haemost 1998; 79: 602-8.
10 Stringer HA, Pannekoek H. The significance of fibrin binding by plasminogen activator inhibitor 1 for the mechanism of tissue-type plasminogen activator-mediated fibrinolysis. J Biol Chem 1995; 270: 11205-8.
11 Pósán E, Ujj G, Kiss A, Telek B, R(k K, Udvardy M. Reduced in vitro clot lysis and release of more active platelet PAI-1 in polycythemia vera and essential thrombocythemia. Thromb Res 1998; 90: 51-6.
12 Zhu Y, Carmeliet P, Fay WP. Plasminogen activator inhibitor-1 is a major determinant of arterial thrombolysis resistance. Circulation 1999; 99: 3050-5.
13 Levi M, Biemond BJ, van Zonneveld AJ, ten Cate JW, Pannekoek H. Inhibition of plasminogen activator inhibitor-1 activity results in promotion of endogenous thrombolysis and inhibition of thrombus extension in models of experimental thrombosis. Circulation 1992; 85: 305-12.
14 Friederich PW, Levi M, Biemond BJ, Charlton P, Templeton D, van Zonneveld AJ, Bevan P, Pannekoek H, ten Cate JW. Novel low-molecular-weight inhibitor of PAI-1 (XR5118) promotes endogenous fibrinolysis and reduces postthrombolysis thrombus growth in rabbits. Circulation 1997; 96: 916-21.
15 Eitzman DT, Fay WP, Lawrence DA, Francis-Chmura AM, Shore JD, Olson ST, Ginsburg D. Peptide – mediated inactivation of recombinant and platelet plasminogen activator inhibitor – 1 in vitro. J Clin Invest 1995; 95: 2416-20.
16 Handt S, Jerome WG, Tietze L, Hantgan RR. Plasminogen activator inhibitor-1 secretion of endothelial cells increases fibrinolytic resistance of an in vitro fibrin clot: evidence for a key role of endothelial cells in thrombolytic resistance. Blood 1996; 87: 4204-13.
17 Pawlowska Z, Pluskota E, Chabielska E, Buczko W, Okruszek A, Stec WJ, Cierniewski CS. Phosphorothioate oligodeoxyribonucleotides antisense to PAI-1 mRNA increase fibrinolysis and modify experimental thrombosis in rats. Thromb Haemost 1998; 79: 348-53.
18 Deutsch DG, Mertz ET. Plasminogen: purification from human plasma by affinity chromatography. Science 1970; 170: 1095-6.
19 Kobylanska A, Pluskota E, Swiatkowska M, Wojcik M, Cierniewska-Cieslak A, Krakowiak A, Boczkowska M, Pawlowska Z, Okruszek A, Koziolkiewicz M, Cierniewski C, Stec WJ. Inhibition of PAI-1 release in EA.hy 926 cell cultures by 5′-end lipophilic modification of oligodeoxyribonucleotides. Acta Biochim Polon 1999; 46: 679-91.
20 Hornstra G, Vendelmans-Starrenburg A. Induction of experimental arterial occlusive thrombi in rats. Atherosclerosis 1973; 17: 369-82.
21 Lewis ML, Nachtwey DS, Damron KL. A miniaturized fibrinolytic assay for plasminogen activators. Thromb Res 1991; 64: 223-34.
22 Dejana E, Villa S, de Gaetano G. Bleeding time in rats: a comparison of different experimental conditions. Thromb Haemost 1982; 48: 108-11.
23 Ikeda K, Nara Y, Yamori Y. Indirect systolic and mean blood pressure determination by a new tail cuff method in spontaneously hypertensive rats. Lab Anim 1991; 25: 26-9.
24 Krieg AM, Tonkinson J, Matson S, Zhao Q, Saxon M, Zhang LM, Bhanja U, Yakubov L, Stein CA. Modification of antisense phosphodiester oligodeoxynucleotides by a 5′ cholesteryl moiety increases cellular association and improves efficacy. Proc Natl Acad Sci USA 1993; 90: 1048-52.
25 Kobylanska A, Pluskota E, Pawlowska Z, Okruszek A, Cierniewski CS, Stec WJ. The cytotoxicity of anti-PAI-1 oligonucleotides and their conjugates. Nucleosides Nucleotides 1999; 18: 1709-10.
26 Torr-Brown SR, Sobel BE. Attenuation of thrombolysis by release of plasminogen activator inhibitor type-1 from platelets. Thromb Res 1993; 72: 413-21.
27 Kruithof EK, Gudinchet A, Bachmann F. Plasminogen activator inhibitor 1 and plasminogen activator inhibitor 2 in various disease states. Thromb Haemost 1988; 59: 7-12.
28 Wiman B, Hamsten A. The fibrinolytic enzyme system and its role in the etiology of thromboembolic disease. Semin Thromb Hemost 1990; 16: 207-16.
29 Aznar J, Estellés A. Role of plasminogen activator inhibitor type 1 in the pathogenesis of coronary artery diseases. Haemostasis 1994; 24: 243-51.
30 Rocha E, Páramo JA. The relationship between impaired fibrinolysis and coronary heart disease: a role for PAI-1. Fibrinolysis 1994; 8: 294-303.
31 Juhan-Vague I, Alessi MC. Plasminogen activator inhibitor-1 and atherothrombosis. Thromb Haemost 1993; 70: 138-43.
32 Charlton PA, Faint RW, Bent F, Bryans J, Chicarelli-Robinson I, Mackie I, Machin S, Bevan P. Evaluation of a low molecular weight modulator of human plasminogen activator inhibitor-1 activity. Thromb Haemost 1996; 75: 808-15.