Targeted SERPIN (TaSER): A dual-action antithrombotic agent that targets platelets for SERPIN delivery.

Autor: Sanrattana W; CDL Research, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., Smits S; CDL Research, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., Barendrecht AD; CDL Research, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., van Kleef ND; CDL Research, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., El Otmani H; CDL Research, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., Zivkovic M; Van Creveldkliniek, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., Roest M; Synapse Research Institute, Maastricht, The Netherlands.; Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, Maastricht, The Netherlands., Renné T; Institute for Clinical Chemistry and Laboratory Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.; Center for Thrombosis and Hemostasis (CTH), Johannes Gutenberg University Medical Center, Mainz, Germany., Clark CC; CDL Research, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., de Maat S; CDL Research, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., Maas C; CDL Research, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
Jazyk: angličtina
Zdroj: Journal of thrombosis and haemostasis : JTH [J Thromb Haemost] 2022 Feb; Vol. 20 (2), pp. 353-365. Date of Electronic Publication: 2021 Oct 29.
DOI: 10.1111/jth.15554
Abstrakt: Background: Occlusive thrombi are not homogeneous in composition. The core of a thrombus is rich in activated platelets and fibrin while the outer shell contains resting platelets. This core is inaccessible to plasma proteins. We produced a fusion protein (targeted SERPIN-TaSER), consisting of a function-blocking V H H against glycoprotein Ibα (GPIbα) and a thrombin-inhibiting serine protease inhibitor (SERPIN; α1-antitrypsin 355 AIAR 358 ) to interfere with platelet-driven thrombin formation.
Aim: To evaluate the antithrombotic properties of TaSER.
Methods: Besides TaSER, we generated three analogous control variants with either a wild-type antitrypsin subunit, a non-targeting control V H H, or their combination. We investigated TaSER and controls in protease activity assays, (platelet-dependent) thrombin generation assays, and by western blotting. The effects of TaSER on platelet activation and von Willebrand factor (VWF) binding were studied by fluorescence-activated cell sorting, in agglutination studies, and in ATP secretion experiments. We studied the influence of TaSER in whole blood (1) on platelet adhesion on VWF, (2) aggregate formation on collagen, and (3) thrombus formation (after recalcification) on collagen and tissue factor.
Results: TaSER binds platelets and inhibits thrombin activity on the platelet surface. It blocks VWF binding and disassembles platelet agglutinates. TaSER delays tissue factor-triggered thrombin generation and ATP secretion in platelet-rich plasma in a targeted manner. In flow studies, TaSER interferes with platelet adhesion and aggregate formation due to GPIbα blockade and limits thrombus formation due to targeted inhibition of platelet-dependent thrombin activity.
Conclusion: The synergy between the individual properties of TaSER makes it a highly effective antithrombotic agent with possible clinical implications.
(© 2021 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals LLC on behalf of International Society on Thrombosis and Haemostasis.)
Databáze: MEDLINE
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