Rapid and Efficient Stable Gene Transfer to Mesenchymal Stromal Cells Using a Modified Foamy Virus Vector.
Rapid and Efficient Stable Gene Transfer to Mesenchymal Stromal Cells Using a Modified Foamy Virus Vector.
复制标题
DOI：
10.1038/mt.2016.91
复制
发表时间：
2016-08
期刊：
影响因子：
--
通讯作者：
McClure MO
中科院分区：
文献类型：
--
作者：
Sweeney NP;Regan C;Liu J;Galleu A;Dazzi F;Lindemann D;Rupar CA;McClure MO
间充质基质细胞(MSCs)在再生医学中具有广阔的应用前景。稳定的体外基因转移可以改善MSCs的治疗效果和范围，但目前的载体需要多轮转导，包括遗传毒性病毒启动子和/或添加细胞毒性阳离子聚合物以实现有效的转导。我们描述了一种自失活泡沫病毒载体(FVV)，它结合了猕猴泡沫病毒的包膜，并使用了生理启动子，在单轮中有效地转导了小鼠骨髓间充质干细胞(MMSCs)。无论是绿色荧光蛋白还是溶酶体酶--芳基硫酸酯A(ARSA)，转基因都得到了持续高效的表达。FVV转导不影响MSC的特性(表面标志物的表达和分化潜能)，以及在脑室内交付后在小鼠脑内的长期植入和分布。同样，超过95%的人骨髓间充质干细胞(HMSCs)可以使用相同的载体稳定地转导，便于人类应用。本工作描述了可用于mMSCs和hMSCs的最稳定的基因转移载体。
Mesenchymal stromal cells (MSCs) hold great promise for regenerative medicine. Stable ex vivo gene transfer to MSCs could improve the outcome and scope of MSC therapy, but current vectors require multiple rounds of transduction, involve genotoxic viral promoters and/or the addition of cytotoxic cationic polymers in order to achieve efficient transduction. We describe a self-inactivating foamy virus vector (FVV), incorporating the simian macaque foamy virus envelope and using physiological promoters, which efficiently transduces murine MSCs (mMSCs) in a single-round. High and sustained expression of the transgene, whether GFP or the lysosomal enzyme, arylsulphatase A (ARSA), was achieved. Defining MSC characteristics (surface marker expression and differentiation potential), as well as long-term engraftment and distribution in the murine brain following intracerebroventricular delivery, are unaffected by FVV transduction. Similarly, greater than 95% of human MSCs (hMSCs) were stably transduced using the same vector, facilitating human application. This work describes the best stable gene transfer vector available for mMSCs and hMSCs.