Novel GPIb-independent platelet aggregation induced by botrocetin: implications for diagnosis and antithrombotic therapy

	Novel GPIb-independent platelet aggregation induced by botrocetin: implications for diagnosis and antithrombotic therapy
	Shen, CB; Mackeigan, DT; Shoara, AA; Bhoria, P; Zhu, GH; Karakas, D; Ma, WJ; Chen, ZY; Xu, RJ; Slavkovic, S; Zhang, DC; Prifti, V; Liu, ZZ; Cerenzia, EG; Chen, PG; Neves, MAD; Li, HY; Xue, F; Yang, RC; Liu, JL; Lai, R; Li, RH; Ni, HY
	2024
发表期刊	JOURNAL OF THROMBOSIS AND HAEMOSTASIS
ISSN	1538-7933
卷号	22 期号:11 页码:3249-3265
摘要	Background: Snake venom botrocetin facilitates von Willebrand factor (VWF) binding to platelet GPIb alpha and has been widely used for the diagnosis of von Willebrand disease and GPIb-related disorders. Botrocetin is also commonly employed for the development/characterization of antithrombotics targeting the GPIb-VWF axis. Objectives: To explore the alternative receptor(s)/mechanisms that participate in botrocetin-induced platelet aggregation. Methods: The effects of botrocetin on platelet aggregation were examined using platelets from wild-type, VWF- and fibrinogen-deficient, GPIb alpha-deficient, IL4R alpha/GPIb alpha- transgenic, ITGA2B and ITGB3-deficient mice, and Bernard-Soulier syndrome and healthy human samples. Platelet-fibrinogen and platelet-VWF interaction were measured using flow cytometry. GPIb alpha-VWF binding was evaluated utilizing enzyme- linked immunosorbent assay. Botrocetin-alpha IIbP3 and botrocetin-GPIb alpha interactions were measured using enzyme-linked immunosorbent assay and fluorescence anisotropy assays. Heparinized whole blood from healthy donors was examined for thrombus formation and growth in a perfusion chamber. Results: Botrocetin could induce aggregation of platelets from a Bernard-Soulier syndrome patient and GPIb alpha-deficient mice as well as platelets lacking the N-terminal extracellular domain of GPIb alpha. Botrocetin could interact with alpha IIb P 3 and facilitated alpha IIb P 3-VWF interaction independent of GPIb. Botrocetin competitively bound to the ligand-binding domain of activated rather than resting alpha IIb P 3 . Although botrocetininduced platelet aggregation requires VWF, strikingly, in the absence of VWF, botrocetin blocked fibrinogen and other ligand binding to alpha IIb P 3 and inhibited platelet aggregation and thrombus formation. Consistently, recombinant botrocetin defective in VWF binding inhibited alpha IIb P 3- and GPIb-mediated platelet aggregation, spreading, and thrombus formation. Conclusion: Our study provides insights into avoiding the misdiagnosis of GPIb-related disorders and developing botrocetin mutants as potential new antithrombotics that may simultaneously target both alpha IIb P 3 and GPIb alpha.
收录类别	SCI
语种	英语
文献类型	期刊论文
条目标识符	http://ir.kiz.ac.cn/handle/152453/14526
专题	科研部门_天然药物功能蛋白质学科组（赖仞）
推荐引用方式 GB/T 7714	Shen, CB,Mackeigan, DT,Shoara, AA,et al. Novel GPIb-independent platelet aggregation induced by botrocetin: implications for diagnosis and antithrombotic therapy[J]. JOURNAL OF THROMBOSIS AND HAEMOSTASIS,2024,22(11):3249-3265.
APA	Shen, CB.,Mackeigan, DT.,Shoara, AA.,Bhoria, P.,Zhu, GH.,...&Ni, HY.(2024).Novel GPIb-independent platelet aggregation induced by botrocetin: implications for diagnosis and antithrombotic therapy.JOURNAL OF THROMBOSIS AND HAEMOSTASIS,22(11),3249-3265.
MLA	Shen, CB,et al."Novel GPIb-independent platelet aggregation induced by botrocetin: implications for diagnosis and antithrombotic therapy".JOURNAL OF THROMBOSIS AND HAEMOSTASIS 22.11(2024):3249-3265.

条目包含的文件
文件名称/大小	文献类型	版本类型	开放类型	使用许可
LR2025040204.pdf（5453KB）	期刊论文	出版稿	开放获取	CC BY-NC-SA	请求全文