A long-acting PAI-1 inhibitor reduces thrombus formation

doi:10.1160/TH16-11-0891

QIBEBT-IR > 蛋白质设计研究组

	A long-acting PAI-1 inhibitor reduces thrombus formation
	Peng, Shuangzhou 1,2,3; Xue, Guangpu 1,2,6; Gong, Lihu 1,2; Fang, Chao 7,8; Chen, Jingfei 5; Yuan, Cai 4; Chen, Zhuo 1,2; Yao, Lishan 5; Furie, Bruce 7; Huang, Mingdong 1,2,4,7
	2017-07-01
发表期刊	THROMBOSIS AND HAEMOSTASIS
卷号	117 期号:7 页码:1338-1347
摘要	Plasminogen activator inhibitor 1 (PAI-1) is the main inhibitor of tissue-type and urokinase-type plasminogen activators (t/uPA) and plays an important role in fibrinolysis. Inhibition of PAI-1 activity prevents thrombosis and accelerates fibrinolysis, indicating that PAI-1 inhibitors may be used as effective antithrombotic agents. We previously designed a PAI-1 inhibitor (PAltrap) which is a variant of inactivated urokinase protease domain. In the present study, we fused PAltrap with human serum albumin (HSA) to develop a long-acting PAI-1 inhibitor. Unfortunately, the fusion protein PAltrap-HSA lost some potency compared to PAltrap (33 nM vs 10 nM). Guided by computational method, we carried out further optimisation to enhance inhibitory potency for PAI-1. The new PAltrap, denominated PAltrap(H37R)-HSA, which was the H37R variant of PAltrap fused to NSA, gave a six-fold improvement of IC50 (5 nM) for human active PAI-1 compared to PAIt-rap-NSA, and showed much longer plasma half-life (200-fold) compared to PAltrap. We further demonstrated that the PAltrap(H37R)-1 HSA inhibited exogenous or endogenous PAI-1 to promote fibrinolysis in fibrin-clot lysis assay. PAltrap(H37R)-HSA inhibits murine PAI-1 with IC50 value of 12 nM, allowing the inhibitor to be evaluated in murine models. Using an intravital microscopy, we demonstrated that PAIt-rap(H37R)-HSA blocks thrombus formation and platelet accumulation in vivo in a laser-induced vascular injury mouse model. Additionally, mouse tail bleeding assay showed that PAltrap(H37R)-HSA did not affect the global haemostasis. These results suggest that PAltrap(H37R)HSA have the potential benefit to prevent thrombosis and accelerates fibrinolysis.
文章类型	Article
关键词	Pai-1 Inhibitor Human Serum Albumin (Hsa) Thrombosis Clot Lysis
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
DOI	10.1160/TH16-11-0891
关键词[WOS]	PLASMINOGEN-ACTIVATOR INHIBITOR-1 ; ARTERIAL THROMBOSIS ; IN-VIVO ; MICE ; THROMBOLYSIS ; STABILITY ; GENE ; FIBRINOLYSIS ; DEFICIENCY ; ANTAGONIST
收录类别	SCI
语种	英语
WOS研究方向	Hematology ; Cardiovascular System & Cardiology
项目资助者	Natural Science Foundation of China(31370737 ; Strategic Priority Research Program of the CAS(XDA09030307) ; CAS/SAFEA International Partnership Program for Creative Research Teams ; Thrombosis and Hemostasis Societies of North America ; 31400637 ; 31570745 ; 31670739 ; U1405229)
WOS类目	Hematology ; Peripheral Vascular Disease
WOS记录号	WOS:000404671700014
引用统计
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/9637
专题	蛋白质设计研究组
作者单位	1.Chinese Acad Sci, Fujian Inst Res Struct Matter, State Key Lab Struct Chem, Fuzhou, Fujian, Peoples R China 2.Chinese Acad Sci, Danish Chinese Ctr Proteases & Canc, Fujian Inst Res Struct Matter, Fuzhou, Fujian, Peoples R China 3.Fujian Agr & Forestry Univ, Coll Life Sci, Fuzhou, Fujian, Peoples R China 4.Fuzhou Univ, Fuzhou, Fujian, Peoples R China 5.Chinese Acad Sci, Qingdao Inst Biomass Energy & Bioproc Technol, Qingdao, Peoples R China 6.Fujian Normal Univ, Coll Life Sci, Fuzhou, Fujian, Peoples R China 7.Harvard Med Sch, Beth Israel Deaconess Med Ctr, Div Hemostasis & Thrombosis, Boston, MA USA 8.Huazhong Univ Sci &Technol, Tongji Med Coll, Sch Basic Med, Huazhong, Peoples R China
推荐引用方式 GB/T 7714	Peng, Shuangzhou,Xue, Guangpu,Gong, Lihu,et al. A long-acting PAI-1 inhibitor reduces thrombus formation[J]. THROMBOSIS AND HAEMOSTASIS,2017,117(7):1338-1347.
APA	Peng, Shuangzhou.,Xue, Guangpu.,Gong, Lihu.,Fang, Chao.,Chen, Jingfei.,...&Huang, Mingdong.(2017).A long-acting PAI-1 inhibitor reduces thrombus formation.THROMBOSIS AND HAEMOSTASIS,117(7),1338-1347.
MLA	Peng, Shuangzhou,et al."A long-acting PAI-1 inhibitor reduces thrombus formation".THROMBOSIS AND HAEMOSTASIS 117.7(2017):1338-1347.