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Thromb Haemost 1994; 71(01): 124-128
DOI: 10.1055/s-0038-1642395
Review Article
Schattauer GmbH Stuttgart

Inhibitor-Resistant Tissue-Type Plasminogen Activator: An Improved Thrombolytic Agent In Vitro

R V Shohet
1   The Department of Internal Medicine, University of Texas, Southwestern Medical School, Dallas, TX, USA
,
S Spitzer
2   The Department of Biochemistry, University of Texas, Southwestern Medical School, Dallas, TX, USA
,
E L Madison
1   The Department of Internal Medicine, University of Texas, Southwestern Medical School, Dallas, TX, USA
,
R Bassel-Duby
1   The Department of Internal Medicine, University of Texas, Southwestern Medical School, Dallas, TX, USA
,
M-J Gething
2   The Department of Biochemistry, University of Texas, Southwestern Medical School, Dallas, TX, USA
,
J F Sambrook
2   The Department of Biochemistry, University of Texas, Southwestern Medical School, Dallas, TX, USA
› Author Affiliations
Further Information

Publication History

Received: 15 March 1993

Accepted after revision 21 September 1993

Publication Date:
12 July 2018 (online)

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Summary

Platelet-rich clots are inefficiently lysed by current fibrinolytic agents. Platelets contain a great deal of plasminogen activator inhibitor 1 (PAI-1), the principal endogenous inhibitor of tissue-type plasminogen activator (t-PA). We have tested whether PAI-1 resistant t-PAs would be more effective thrombolytic agents in an in vitro model of platelet rich clots. Clots were formed with recalcified human plasma without or with the addition of platelets. The lysis of these clots was followed by the release of incorporated 125I-fibrinogen. Mutant and wild-type t-PA were almost equally effective against clots lacking platelets but the mutant was twice as effective at lysing platelet-rich clots. A mechanism for this effect is suggested by the demonstration that a complex between wild-type t-PA and extruded platelet contents resembles that between purified t-PA and PAI-1 and that the PAI-1 resistant t-PA does not interfere with formation of this adduct. Because of its enhanced ability to lyse platelet-rich clots in vitro, further in vivo work may find that PAI-1 resistant t-PA is a more efficacious therapeutic agent than wild-type t-PA in situations where platelets contribute to the failure of thrombolysis.

 

  • References

  • 1 Madison EL, Goldsmith EJ, Gerard RD, Gething M-JH, Sambrook JF. Serpin-resistant mutants of human tissue-type plasminogen activator. Nature 1989; 339: 721-4
    CrossrefPubMedGoogle Scholar
  • 2 Chesbro JH. and the TIMI investigators. Thrombolysis in myocardial infarction (TIMI) trial, phase 1: a comparison between intravenous tissue plasminogen activator and intravenous streptokinase. Circulation 1987; 76: 142-54
    CrossrefPubMedGoogle Scholar
  • 3 Sherry S, Marder VJ. Streptokinase and recombinant tissue plasminogen activator (rt-PA) are equally effective in treating acute myocardial infarction. Ann Int Med 1991; 114: 417-23
    CrossrefPubMedGoogle Scholar
  • 4 Hoylaerts M, Rijken DC, Lijnen HR, Collen D. Kinetics of the activation of plasminogen by human tissue plasminogen activator. Role of fibrin. J Biol Chern 1982; 257: 2912-9
    PubMedGoogle Scholar
  • 5 Kennedy JW, Ritchie JL, Davis KB, Stadius ML, Maynard C, Fritz JK. The Western Washington randomized trial of intracoronary streptokinase in acute myocardial infarction. N Engl J Med 1985; 312: 1073-8
    CrossrefPubMedGoogle Scholar
  • 6 Cigarroa RC, Lange RA, Hillis LD. Prognosis after acute myocardial infarction in patients with and without residual anterograde coronary blood flow. Am J Cardiol 1989; 64: 155-60
    CrossrefPubMedGoogle Scholar
  • 7 Hirsh J, Buchanan MR, Ofosu FA, Weitz J. Evolution of thrombosis. Ann NY Acad Sci 1987; 516: 586-604
    CrossrefPubMedGoogle Scholar
  • 8 Walsh PN. Platelet-mediated coagulant protein interactions in hemostasis. Semin Hemat 1985; 22: 178-86
    PubMedGoogle Scholar
  • 9 Fay WP, Owen WG. Platelet plasminogen activator inhibitor: purification and characterization of interaction with plasminogen activators and activated protein C. Biochemistry 1989; 28: 5773-8
    CrossrefPubMedGoogle Scholar
  • 10 Wiman B, Chmielewska J, Ranby M. Inactivation of tissue plasminogen activator in plasma. J Biol Chem 1984; 259: 3644-7
    PubMedGoogle Scholar
  • 11 Jang I-K, Gold HK, Ziskind AA, Fallon JT, Holt RE, Leinbach RC, May JW, Collen D. Differential sensitivity of erythrocyte-rich and platelet-rich arterial thrombi to lysis with recombinant tissue-type plasminogen activator. Circulation 1989; 79: 920-8
    CrossrefPubMedGoogle Scholar
  • 12 Yasuda T, Gold HK, Leinbach RC, Saito T, Guerrero JL, Jang I-K, Holt R, Fallon JT, Collen D. Lysis of plasminogen activator-resistant platelet-rich coronary artery thrombosis with combined bolus injection of recombinant tissue-type plasminogen activator and antiplatelet GPIIb/IIIa antibody. J Amer Coll Cardiol 1990; 16: 1728-35
    CrossrefPubMedGoogle Scholar
  • 13 Bassel-Duby R, Jiang NY, Bittick T, Madison E, Mc Gookey D, Orth K, Shohet R, Sambrook J, Gething M-J. Tyrosine-67 in the epidermal growth factor-like domain of tissue-type plasminogen activator is important for clearance by a specific hepatic receptor. J Biol Chem 1992; 267: 9668-77
    PubMedGoogle Scholar
  • 14 Boose JA, Kuismanen E, Gerard R, Sambrook J, Gething M-J. The single-chain form of tissue-type plasminogen activator has catalytic activity: studies with a mutant enzyme that lacks the cleavage site. Biochemistry 1989; 28: 635-43
    CrossrefPubMedGoogle Scholar
  • 15 Larsen GR, Henson K, Blue Y. Variants of human tissue-type plasminogen activator. J Biol Chem 1988; 263: 1023-9
    PubMedGoogle Scholar
  • 16 Lund LF, Georg B, Nielsen LS, Mayer M, Dano K, Andreasen P. Plasminogen activator inhibitor type 1: cell-specific and differentiation-induced expression and regulation in human cell lines, as determined by enzyme-linked immunosorbent assay. Molec and Cell Endo 1988; 60: 43-53
    PubMedGoogle Scholar
  • 17 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5
    CrossrefPubMedGoogle Scholar
  • 18 Clauss VA. Gerinnungsphysiologische Schnellmethode zur Bestimmung des Fibrinogens. Acta Haemat 1957; 17: 237-46
    CrossrefPubMedGoogle Scholar
  • 19 Madison EL, Goldsmith EJ, Gerard RD, Gething M-JH, Sambrook JF, Bassel-Duby RS. Amino acid residues that affect interaction of tissue-type plasminogen activator with plasminogen activator inhibitor 1. Proc Natl Acad Sci USA 1990; 87: 3530-3
    CrossrefPubMedGoogle Scholar
  • 20 Maynard C, Althouse R, Olsufka M, Ritchie JL, Davis KB, Kennedy JW. Early versus late hospital arrival for acute myocardial infarction in the Western Washington thrombolytic therapy trials. Am J Cardiol 1989; 63: 1296-300
    CrossrefPubMedGoogle Scholar
  • 21 Li X-K, Lijnen HR, Nelles L, Van Hoef B, Stassen JM, Collen D. Biochemical and biologic properties of rt-PA del (K296-G302), a recombinant human tissue-type plasminogen activator deletion mutant resistant to plasminogen activator inhibitor-1. Blood 1992; 79: 417-29
    PubMedGoogle Scholar