Dataset Information

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

ABSTRACT:

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 AIAR³⁵⁸ ) 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.

SUBMITTER: Sanrattana W

PROVIDER: S-EPMC9298318 | biostudies-literature |

REPOSITORIES: biostudies-literature

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