Dual roles of fucoidan-GPIbα interaction in thrombosis and hemostasis: implications for drug development targeting GPIbα

Chuanbin Shen; Daniel T Mackeigan; Aron A Shoara; Runjia Xu; Preeti Bhoria; Danielle Karakas; Wenjing Ma; Eric Cerenzia; ZiYan Chen; Brock Hoard; Lisha Lin; Xi Lei; Guangheng Zhu; Pingguo Chen; Philip E Johnson; Heyu Ni

doi:10.1016/j.jtha.2022.12.030

Dual roles of fucoidan-GPIbα interaction in thrombosis and hemostasis: implications for drug development targeting GPIbα

J Thromb Haemost. 2023 May;21(5):1274-1288. doi: 10.1016/j.jtha.2022.12.030. Epub 2023 Jan 9.

Authors

Chuanbin Shen¹, Daniel T Mackeigan², Aron A Shoara³, Runjia Xu³, Preeti Bhoria⁴, Danielle Karakas³, Wenjing Ma⁴, Eric Cerenzia², ZiYan Chen³, Brock Hoard³, Lisha Lin⁵, Xi Lei⁶, Guangheng Zhu⁶, Pingguo Chen⁷, Philip E Johnson⁸, Heyu Ni⁹

Affiliations

¹ Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, M5S 1A1, ON, Canada; Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Shandong, China; Department of Laboratory Medicine, LKSKI-Keenan Research Centre for Biomedical Science, St. Michael's Hospital, and Toronto Platelet Immunobiology Group, Toronto, Canada.
² Department of Laboratory Medicine, LKSKI-Keenan Research Centre for Biomedical Science, St. Michael's Hospital, and Toronto Platelet Immunobiology Group, Toronto, Canada; Department of Physiology, University of Toronto, Toronto, Canada.
³ Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, M5S 1A1, ON, Canada; Department of Laboratory Medicine, LKSKI-Keenan Research Centre for Biomedical Science, St. Michael's Hospital, and Toronto Platelet Immunobiology Group, Toronto, Canada.
⁴ Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, M5S 1A1, ON, Canada; Department of Laboratory Medicine, LKSKI-Keenan Research Centre for Biomedical Science, St. Michael's Hospital, and Toronto Platelet Immunobiology Group, Toronto, Canada; CCOA Therapeutics Inc Toronto, Canada.
⁵ Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, M5S 1A1, ON, Canada; Department of Laboratory Medicine, LKSKI-Keenan Research Centre for Biomedical Science, St. Michael's Hospital, and Toronto Platelet Immunobiology Group, Toronto, Canada; State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China.
⁶ Department of Laboratory Medicine, LKSKI-Keenan Research Centre for Biomedical Science, St. Michael's Hospital, and Toronto Platelet Immunobiology Group, Toronto, Canada; CCOA Therapeutics Inc Toronto, Canada.
⁷ Department of Laboratory Medicine, LKSKI-Keenan Research Centre for Biomedical Science, St. Michael's Hospital, and Toronto Platelet Immunobiology Group, Toronto, Canada; Canadian Blood Services Centre for Innovation, Toronto, Canada.
⁸ Department of Chemistry and Centre for Research on Biomolecular Interactions, York University, Toronto, Canada.
⁹ Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, M5S 1A1, ON, Canada; Department of Laboratory Medicine, LKSKI-Keenan Research Centre for Biomedical Science, St. Michael's Hospital, and Toronto Platelet Immunobiology Group, Toronto, Canada; Department of Physiology, University of Toronto, Toronto, Canada; CCOA Therapeutics Inc Toronto, Canada; Canadian Blood Services Centre for Innovation, Toronto, Canada; Department of Medicine, University of Toronto, Toronto, Canada. Electronic address: nih@smh.ca.

PMID: 36732162
DOI: 10.1016/j.jtha.2022.12.030

Abstract

Background: Platelet GPIbα-von Willebrand factor (VWF) interaction initiates platelet adhesion, activation, and thrombus growth, especially under high shear conditions. Therefore, the GPIb-VWF axis has been suggested as a promising target against arterial thrombosis. The polysaccharide fucoidan has been reported to have opposing prothrombotic and antithrombotic effects; however, its binding mechanism with platelets has not been adequately studied.

Objective: The objective of this study was to explore the mechanism of fucoidan and its hydrolyzed products in thrombosis and hemostasis.

Methods: Natural fucoidan was hydrolyzed by using hydrochloric acid and was characterized by using size-exclusion chromatography, UV-visible spectroscopy, and fluorometry techniques. The effects of natural and hydrolyzed fucoidan on platelet aggregation were examined by using platelets from wild-type, VWF and fibrinogen-deficient, GPIbα-deficient, and IL4Rα/GPIbα-transgenic and αIIb-deficient mice and from human beings. Platelet activation markers (P-selectin expression, PAC-1, and fibrinogen binding) and platelet-VWF A1 interaction were measured by using flow cytometry. GPIbα-VWF A1 interaction was evaluated by using enzyme-linked immunosorbent assay. GPIb-IX-induced signal transduction was detected by using western blot. Heparinized whole blood from healthy donors was used to test thrombus formation and growth in a perfusion chamber.

Results: We found that GPIbα is critical for fucoidan-induced platelet activation. Fucoidan interacted with the extracellular domain of GPIbα and blocked its interaction with VWF but itself could lead to GPIbα-mediated signal transduction and, subsequently, αIIbβ3 activation and platelet aggregation. Conversely, low-molecular weight fucoidan inhibited GPIb-VWF-mediated platelet aggregation, spreading, and thrombus growth at high shear.

Conclusion: Fucoidan-GPIbα interaction may have unique therapeutic potential against bleeding disorders in its high-molecular weight state and protection against arterial thrombosis by blocking GPIb-VWF interaction after fucoidan is hydrolyzed.

Keywords: fucoidan; glycoprotein Ib; platelet; thrombosis; von Willebrand factor.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Blood Platelets / metabolism
Fibrinogen / metabolism
Humans
Mice
Platelet Aggregation
Platelet Glycoprotein GPIb-IX Complex / metabolism
Polysaccharides / pharmacology
Protein Binding
Thrombosis* / drug therapy
Thrombosis* / metabolism
Thrombosis* / prevention & control
von Willebrand Factor* / metabolism

Substances

fucoidan
von Willebrand Factor
Platelet Glycoprotein GPIb-IX Complex
Polysaccharides
Fibrinogen

Grants and funding

389035/CIHR/Canada