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糖蛋白VI（GPVI）或GPVI-CD39包被的纳米颗粒的表征及其对体外血栓形成的影响。

Characterization of GPVI- or GPVI-CD39-Coated Nanoparticles and Their Impact on In Vitro Thrombus Formation.

作者信息

Nestele Jeremy A, Rohlfing Anne-Katrin, Dicenta Valerie, Bild Alexander, Eißler Daniela, Emschermann Frederic, Kremser Marcel, Krutzke Konstantin, Schäffer Tilman E, Borst Oliver, Levi Moran, Korin Netanel, Gawaz Meinrad Paul

机构信息

Department of Cardiology and Angiology, University of Tübingen, 72076 Tubingen, Germany.

DFG Heisenberg Group Thrombocardiology, University of Tübingen, 72076 Tubingen, Germany.

出版信息

Int J Mol Sci. 2021 Dec 21;23(1):11. doi: 10.3390/ijms23010011.

DOI:10.3390/ijms23010011
PMID:35008437
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8744670/
Abstract

Traditional antithrombotic agents commonly share a therapy-limiting side effect, as they increase the overall systemic bleeding risk. A novel approach for targeted antithrombotic therapy is nanoparticles. In other therapeutic fields, nanoparticles have enabled site-specific delivery with low levels of toxicity and side effects. Here, we paired nanotechnology with an established dimeric glycoprotein VI-Fc (GPVI-Fc) and a GPVI-CD39 fusion protein, thereby combining site-specific delivery and new antithrombotic drugs. Poly(lactic-co-glycolic acid) (PLGA) nanoparticles, NP-BSA, NP-GPVI and NP-GPVI-CD39 were characterized through electron microscopy, atomic force measurements and flow cytometry. Light transmission aggregometry enabled analysis of platelet aggregation. Thrombus formation was observed through flow chamber experiments. NP-GPVI and NP-GPVI-CD39 displayed a characteristic surface coating pattern. Fluorescence properties were identical amongst all samples. NP-GPVI and NP-GPVI-CD39 significantly impaired platelet aggregation. Thrombus formation was significantly impaired by NP-GPVI and was particularly impaired by NP-GPVI-CD39. The receptor-coated nanoparticles NP-GPVI and the bifunctional molecule NP-GPVI-CD39 demonstrated significant inhibition of in vitro thrombus formation. Consequently, the nanoparticle-mediated antithrombotic effect of GPVI-Fc, as well as GPVI-CD39, and an additive impact of CD39 was confirmed. In conclusion, NP-GPVI and NP-GPVI-CD39 may serve as a promising foundation for a novel therapeutic approach regarding targeted antithrombotic therapy.

摘要

传统抗血栓药物通常都有一个限制治疗的副作用，即它们会增加整体系统性出血风险。纳米颗粒是一种靶向抗血栓治疗的新方法。在其他治疗领域，纳米颗粒能够实现低毒性和副作用的位点特异性递送。在此，我们将纳米技术与已有的二聚体糖蛋白VI-Fc（GPVI-Fc）和GPVI-CD39融合蛋白相结合，从而将位点特异性递送与新型抗血栓药物结合起来。通过电子显微镜、原子力测量和流式细胞术对聚乳酸-乙醇酸共聚物（PLGA）纳米颗粒、NP-BSA、NP-GPVI和NP-GPVI-CD39进行了表征。光透射聚集法可用于分析血小板聚集情况。通过流动腔实验观察血栓形成。NP-GPVI和NP-GPVI-CD39呈现出特征性的表面涂层模式。所有样品的荧光特性相同。NP-GPVI和NP-GPVI-CD39显著抑制血小板聚集。NP-GPVI显著抑制血栓形成，NP-GPVI-CD39对血栓形成的抑制作用尤为明显。受体包被的纳米颗粒NP-GPVI和双功能分子NP-GPVI-CD39对体外血栓形成具有显著抑制作用。因此，证实了纳米颗粒介导的GPVI-Fc以及GPVI-CD39的抗血栓作用，以及CD39的相加作用。总之，NP-GPVI和NP-GPVI-CD39可能为靶向抗血栓治疗的新型治疗方法提供一个有前景的基础。

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