Stable thrombus formation on irradiated microvascular endothelial cells under pulsatile flow: Pre-testing annexin V-thrombin conjugate for treatment of brain arteriovenous malformations
| Autor: | Zhenjun Zhao, Lucinda S. McRobb, Valery Combes, Vivienne S. Lee, Andrew J. Gauden, S.O. Ugoya, Georges E. Grau, David W. Inglis, Marcus A. Stoodley, Vaughan Moutrie, E.D. Santos, Sinduja Subramanian |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Pathology
medicine.medical_specialty Endothelium 030204 cardiovascular system & hematology Fibrinogen Arteriovenous Malformations 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Thrombin Annexin medicine Humans Platelet Annexin A5 Thrombus Brain Endothelial Cells Thrombosis Hematology Phosphatidylserine medicine.disease medicine.anatomical_structure Cardiovascular System & Hematology chemistry 030217 neurology & neurosurgery medicine.drug Conjugate |
| Zdroj: | Thrombosis Research. 167:104-112 |
| ISSN: | 0049-3848 |
| DOI: | 10.1016/j.thromres.2018.05.016 |
| Popis: | © 2018 Elsevier Ltd Background: Our goal is to develop a vascular targeting treatment for brain arteriovenous malformations (AVMs). Externalized phosphatidylserine has been established as a potential biomarker on the endothelium of irradiated AVM blood vessels. We hypothesize that phosphatidylserine could be selectively targeted after AVM radiosurgery with a ligand-directed vascular targeting agent to achieve localized thrombosis and rapid occlusion of pathological AVM vessels. Objective: The study aim was to establish an in vitro parallel-plate flow chamber to test the efficacy of a pro-thrombotic conjugate targeting phosphatidylserine. Methods: Conjugate was prepared by Lys-Lys cross-linking of thrombin with the phosphatidylserine-targeting ligand, annexin V. Cerebral microvascular endothelial cells were irradiated (5, 15, and 25 Gy) and after 1 or 3 days assembled in a parallel-plate flow chamber containing whole human blood and conjugate (1.25 or 2.5 μg/mL). Confocal microscopy was used to assess thrombus formation after flow via binding and aggregation of fluorescently-labelled platelets and fibrinogen. Results and conclusions: The annexin V-thrombin conjugate induced rapid thrombosis (fibrin deposition) on irradiated endothelial cells under shear stress in the parallel-plate flow device. Unconjugated, non-targeting thrombin did not induce fibrin deposition. A synergistic interaction between radiation and conjugate dose was observed. Thrombosis was greatest at the highest combined doses of radiation (25 Gy) and conjugate (2.5 μg/mL). The parallel-plate flow system provides a rapid method to pre-test pro-thrombotic vascular targeting agents. These findings validate the translation of the annexin V-thrombin conjugate to pre-clinical studies. |
| Databáze: | OpenAIRE |
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