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Robust thrombolytic and anti-inflammatory action of a constitutively active ADAMTS13 variant in murine stroke models.
Blood ( IF 20.3 ) Pub Date : 2022-03-10 , DOI: 10.1182/blood.2021012787
Kieron South 1, 2 , Ohud Saleh 1, 2, 3 , Eloise Lemarchand 1, 2 , Graham Coutts 1, 2 , Craig J Smith 2, 4, 5 , Ingo Schiessl 1, 2 , Stuart M Allan 1, 2
Affiliation  

Advances in our understanding of ADAMTS13 structure, and the conformation changes required for full activity, have rejuvenated the possibility of its use as a thrombolytic therapy. We have tested a novel Ala1144Val ADAMTS13 variant (constitutively active [ca] ADAMTS13) that exhibits constitutive activity, characterized using in vitro assays of ADAMTS13 activity, and greatly enhanced thrombolytic activity in 2 murine models of ischemic stroke, the distal FeCl3 middle cerebral artery occlusion (MCAo) model and transient middle cerebral artery occlusion (tMCAO) with systemic inflammation and ischemia/reperfusion injury. The primary measure of efficacy in both models was restoration of regional cerebral blood flow (rCBF) to the MCA territory, which was determined using laser speckle contrast imaging. The caADAMTS13 variant exhibited a constitutively active conformation and a fivefold enhanced activity against fluorescence resonance energy transfer substrate von Willebrand factor 73 (FRETS-VWF73) compared with wild-type (wt) ADAMTS13. Moreover, caADAMTS13 inhibited VWF-mediated platelet capture at subphysiological concentrations and enhanced t-PA/plasmin lysis of fibrin(ogen), neither of which were observed with wtADAMTS13. Significant restoration of rCBF and reduced lesion volume was observed in animals treated with caADAMTS13. When administered 1 hour after FeCl3 MCAo, the caADAMTS13 variant significantly reduced residual VWF and fibrin deposits in the MCA, platelet aggregate formation, and neutrophil recruitment. When administered 4 hours after reperfusion in the tMCAo model, the caADAMTS13 variant induced a significant dissolution of platelet aggregates and a reduction in the resulting tissue hypoperfusion. The caADAMTS13 variant represents a potentially viable therapeutic option for the treatment of acute ischemic stroke, among other thrombotic indications, due to its enhanced in vitro and in vivo activities that result from its constitutively active conformation.

中文翻译:

组成型活性 ADAMTS13 变体在小鼠中风模型中的强大溶栓和抗炎作用。

我们对 ADAMTS13 结构以及完全活性所需的构象变化的理解取得了进展,使其重新用作溶栓疗法的可能性。我们已经测试了一种新的 Ala1144Val ADAMTS13 变体(组成型活性 [ca] ADAMTS13),它表现出组成型活性,使用 ADAMTS13 活性的体外测定来表征,并在 2 种缺血性中风小鼠模型中显着增强溶栓活性,远端 FeCl3 大脑中动脉闭塞(MCAo) 模型和短暂的大脑中动脉闭塞 (tMCAO) 与全身炎症和缺血/再灌注损伤。两种模型中疗效的主要衡量标准是将局部脑血流量 (rCBF) 恢复到 MCA 区域,这是使用激光散斑对比成像确定的。与野生型 (wt) ADAMTS13 相比,caADAMTS13 变体表现出组成型活性构象和对荧光共振能量转移底物血管性血友病因子 73 (FRETS-VWF73) 的五倍增强的活性。此外,caADAMTS13 在亚生理浓度下抑制 VWF 介导的血小板捕获并增强 t-PA/纤溶酶对纤维蛋白(原)的溶解,而 wtADAMTS13 均未观察到这两种情况。在用 caADAMTS13 治疗的动物中观察到 rCBF 显着恢复和病变体积减少。在 FeCl3 MCAo 后 1 小时给药时,caADAMTS13 变体显着减少了 MCA 中残留的 VWF 和纤维蛋白沉积、血小板聚集体形成和中性粒细胞募集。在 tMCAo 模型中再灌注后 4 小时给药时,caADAMTS13 变体诱导了血小板聚集体的显着溶解并减少了由此产生的组织低灌注。caADAMTS13 变体代表了治疗急性缺血性中风以及其他血栓适应症的潜在可行的治疗选择,因为其组成型活性构象导致其增强的体外和体内活性。
更新日期:2021-11-15
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