Molecular requirements for safer generation of thrombolytics by bioengineering the tissue-type plasminogen activator A chain

Authors

  • J. Parcq,

    1. Inserm, Inserm UMR-S U919, University of Caen Basse-Normandie, Serine Proteases and Pathophysiology of the Neurovascular Unit (SP2U), GIP Cyceron, Caen, France
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  • T. Bertrand,

    1. Inserm, Inserm UMR-S U919, University of Caen Basse-Normandie, Serine Proteases and Pathophysiology of the Neurovascular Unit (SP2U), GIP Cyceron, Caen, France
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  • A. F. Baron,

    1. Inserm, Inserm UMR-S U919, University of Caen Basse-Normandie, Serine Proteases and Pathophysiology of the Neurovascular Unit (SP2U), GIP Cyceron, Caen, France
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  • Y. Hommet,

    1. Inserm, Inserm UMR-S U919, University of Caen Basse-Normandie, Serine Proteases and Pathophysiology of the Neurovascular Unit (SP2U), GIP Cyceron, Caen, France
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  • E. Anglès-Cano,

    1. Inserm, Inserm UMR-S U919, University of Caen Basse-Normandie, Serine Proteases and Pathophysiology of the Neurovascular Unit (SP2U), GIP Cyceron, Caen, France
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  • D. Vivien

    Corresponding author
    • Inserm, Inserm UMR-S U919, University of Caen Basse-Normandie, Serine Proteases and Pathophysiology of the Neurovascular Unit (SP2U), GIP Cyceron, Caen, France
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Correspondence: Denis Vivien, Inserm, Inserm UMR-S U919, Université Caen Basse-Normandie, GIP Cyceron, Bd H. Becquerel, BP 5229, Caen, F-14074 France.

Tel.: +33 2 31 47 01 66; fax: +33 2 31 47 02 22.

E-mail: vivien@cyceron.fr

Summary

Background

Thrombolysis with tissue-type plasminogen activator (t-PA) is the only treatment approved for acute ischemic stroke. Although t-PA is an efficient clot lysis enzyme, it also causes damage to the neurovascular unit, including hemorrhagic transformations and neurotoxicity.

Objectives

On the basis of the mechanism of action of t-PA on neurotoxicity, we aimed at studying the molecular requirements to generate safer thrombolytics.

Methods

We produced original t-PA-related mutants, including a non-cleavable single-chain form with restored zymogenicity (sc*-t-PA) and a t-PA modified in the kringle 2 lysine-binding site (K2*-t-PA). Both sc*-t-PA and K2*-t-PA showed fibrinolytic activities similar to that of wild-type t-PA on both euglobulin-containing and plasma-containing clots. In contrast to wild-type t-PA, the two mutants did not promote N-methyl-d-aspartate receptor-mediated neurotoxicity.

Conclusions

We designed t-PA mutants with molecular properties that, in contrast to t-PA, do not induce neurotoxicity.

Ancillary