Research Division of Cell and Molecular Medicine, Center for Molecular Medicine, Jichi Medical University, Tochigi, Japan.
J Thromb Haemost. 2012 Sep;10(9):1802-13. doi: 10.1111/j.1538-7836.2012.04851.x.
Transplantation of cells overexpressing a target protein represents a viable gene therapeutic approach for treating hemophilia. Here, we focused on the use of autologous mesenchymal stem cells (MSCs) expressing coagulation factor for the treatment of coagulation factor VIII (FVIII) deficiency in mice.
Analysis of luciferase gene constructs driven by different promoters revealed that the plasminogen activator inhibitor-1 (PAI-1) gene promoter coupled with the cytomegalovirus promoter enhancer region was one of the most effective promoters for producing the target protein. MSCs transduced with the simian immunodeficiency virus (SIV) vector containing the FVIII gene driven by the PAI-1 promoter expressed FVIII for several months, and this expression was maintained after multiple mesenchymal lineage differentiation. Although intravenous injection of cell supernatant derived from MSCs transduced with an SIV vector containing the FVIII gene driven by the PAI-1 promoter significantly increased plasma FVIII levels, subcutaneous implantation of the MSCs resulted in a transient and weak increase in plasma FVIII levels in FVIII-deficient mice. Interestingly, intra-articular injection of the transduced MSCs significantly ameliorated the hemarthrosis and hemophilic arthropathy induced by knee joint needle puncture in FVIII-deficient mice. The therapeutic effects of a single intra-articular injection of transduced MSCs to inhibit joint bleeding persisted for at least 8 weeks after administration.
MSCs provide a promising autologous cell source for the production of coagulation factor. Intra-articular injection of MSCs expressing coagulation factor may offer an attractive treatment approach for hemophilic arthropathy.
过表达目标蛋白的细胞移植是治疗血友病的一种可行的基因治疗方法。在这里，我们专注于使用表达凝血因子的自体间充质干细胞 (MSC) 来治疗小鼠凝血因子 VIII (FVIII) 缺乏症。
分析不同启动子驱动的荧光素酶基因构建体表明，纤溶酶原激活物抑制剂-1 (PAI-1) 基因启动子与巨细胞病毒启动子增强子区域相连，是产生目标蛋白最有效的启动子之一。用含有由 PAI-1 启动子驱动的 FVIII 基因的 SIV 载体转导的 MSC 可表达 FVIII 长达数月，并且这种表达在多次间充质谱系分化后仍能维持。虽然静脉注射含有由 PAI-1 启动子驱动的 FVIII 基因的 SIV 载体转导的 MSC 的细胞上清液可显著提高血浆 FVIII 水平，但皮下植入 MSC 仅导致 FVIII 缺乏小鼠血浆 FVIII 水平短暂而微弱地增加。有趣的是，关节内注射转导的 MSC 可显著改善 FVIII 缺乏小鼠膝关节针穿刺引起的关节积血和血友病性关节炎。单次关节内注射转导 MSC 抑制关节出血的治疗效果在给药后至少持续 8 周。
MSC 为凝血因子的产生提供了一种有前途的自体细胞来源。关节内注射表达凝血因子的 MSC 可能为血友病性关节炎提供一种有吸引力的治疗方法。
