Safe and efficient local gene delivery into skeletal muscle via a combination of Pluronic L64 and modified electrotransfer
Safe and efficient local gene delivery into skeletal muscle via a combination of Pluronic L64 and modified electrotransfer
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通过 Pluronic L64 和改良电转移的组合，安全高效地将局部基因递送到骨骼肌中
DOI：
10.1038/gt.2014.27
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发表时间：
2014-06-01
期刊：
影响因子：
5.1
通讯作者：
Gu, Z.
中科院分区：
文献类型：
--
作者：
Liu, S.;Ma, L.;Gu, Z.
通过连续组合两种类型的电脉冲可以实现有效的DNA电转移到肌肉中：用于细胞电透化的短高压（HV）脉冲和用于DNA电泳到细胞中的长低压（LV）脉冲。然而，目前施加的电压仍然可以诱导组织的组织学和功能损伤。Pluronic L 64被认为是具有细胞膜干扰能力的分子。因此，我们希望L 64可以作为HV脉冲在细胞膜透化中的替代物，并且安全的LV脉冲仍然可以保持驱动质粒DNA穿过透化膜的能力。在这项工作中，我们优化了电转移参数，以建立一个安全和有效的程序，使用临床应用的仪器，并发现，一个成功的电转移与L 64相结合的关键条件是，质粒/L 64混合物的注射应施加在电转移前1小时。此外，我们发现组合程序无法有效地将质粒转移到实体瘤中，因为未压缩的质粒可能会迅速渗透渗漏的肿瘤血管并流走。总之，结果表明，该组合程序具有通过安全有效的局部基因递送到骨骼肌中的基于质粒的基因治疗的潜力。
Efficient DNA electrotransfer into muscles can be achieved by combining two types of electronic pulses sequentially: short high-voltage (HV) pulse for the cell electropermeabilization and long low-voltage (LV) pulse for the DNA electrophoresis into cells. However, the voltages currently applied can still induce histological and functional damages to tissues. Pluronic L64 has been considered as a molecule possessing cell membrane-disturbing ability. For these reasons, we hope that L64 can be used as a substitute for the HV pulse in cell membrane permeabilization, and a safe LV pulse may still keep the ability to drive plasmid DNA across the permeabilized membrane. In this work, we optimized the electrotransfer parameters to establish a safe and efficient procedure using a clinically applied instrument, and found out that the critical condition for a successful combination of electrotransfer with L64 was that the injection of plasmid/L64 mixture should be applied 1 h before the electrotransfer. In addition, we revealed that the combined procedure could not efficiently transfer plasmid into solid tumor because the uncompressed plasmid may rapidly permeate the leaky tumor vessels and flow away. Altogether, the results demonstrate that the combined procedure has the potential for plasmid-based gene therapy through safe and efficient local gene delivery into skeletal muscles.