Restoration of plasma von Willebrand factor deficiency is sufficient to correct thrombus formation after gene therapy for severe von Willebrand disease
Restoration of plasma von Willebrand factor deficiency is sufficient to correct thrombus formation after gene therapy for severe von Willebrand disease
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DOI：
10.1161/atvbaha.108.168369
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发表时间：
2008-09-01
影响因子：
8.7
通讯作者：
Vanhoorelbeke, Karen
中科院分区：
文献类型：
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作者：
De Meyer, Simon F.;Vandeputte, Nele;Vanhoorelbeke, Karen
目的：基因治疗严重血管性血友病（vWD）似乎是一种有趣的治疗方案，具有长期治疗潜力。我们研究了以肝脏为靶点进行生理活性血管性血友病因子（vWF）异位表达的可行性。方法与结果：在小鼠重度vWD模型中，通过水动力注射，研究了转基因编码的小鼠vWF在肝脏特异性基因转移后恢复vWF功能的能力。通过使用肝细胞特异性α - 1抗胰蛋白酶启动子，与巨细胞病毒启动子相比，获得了更高更持久的vWF表达，达到了比野生型高10 +/- 1倍的最大vWF血浆水平。肝脏表达的vWF显示了全方位的多聚体，包括高分子量多聚体，并恢复了血浆因子VIII水平，这与基因转移后3天（但不是7天）出血时间的纠正一致。重要的是，转基因编码的血浆vWF在FeCl3诱导的血栓形成模型中恢复了适当的血小板粘附和聚集。结论：将转基因编码的血浆vWF基因转入肝脏后，可获得高异位表达。肝脏表达的vWF是完全多聚的，能够在严重的vWD中恢复适当的血小板栓形成。因此，肝脏似乎是严重vWD基因治疗的一个有吸引力的靶点。
Objective-Gene therapy for severe von Willebrand disease (vWD) seems an interesting treatment alternative with long-term therapeutic potential. We investigated the feasibility of targeting the liver for ectopic expression of physiologically active von Willebrand factor (vWF).Methods and Results-The capacity of transgene-encoded murine vWF to restore vWF function was studied in a mouse model of severe vWD after liver-specific gene transfer by hydrodynamic injection. By using a hepatocyte-specific alpha 1 antitrypsin promoter, a considerably higher and longer-lasting vWF expression was obtained when compared with a cytomegalovirus promoter, reaching maximum vWF plasma levels that are 10 +/- 1 times higher than the wild-type level. Liver-expressed vWF showed the full range of multimers, including the high molecular weight multimers, and restored factor VIII plasma levels, consistent with correction of the bleeding time 3 but not 7 days after gene transfer. Importantly, transgene encoded plasma vWF restored proper platelet adhesion and aggregation in a FeCl3 induced thrombosis model.Conclusions-High ectopic expression of transgene encoded plasma vWF can be obtained after gene transfer to the liver. Liver-expressed vWF was fully multimerized and able to restore proper platelet plug formation in severe vWD. The liver therefore seems an attractive target for gene therapy for severe vWD.