Riera Marta, Chillon Miguel, Aran Josep M, Cruzado Josep M, Torras Joan, Grinyó Josep M, Fillat Cristina
Medical and Molecular Genetic Center, Institut de Recerca Oncològica (IRO), Bellaterra, Spain.
J Gene Med. 2004 Jan;6(1):111-8. doi: 10.1002/jgm.463.
The systemic administration of a therapeutic protein is a common approach for the treatment of multiple disorders. Intramuscular (i.m.) injection of plasmids, followed by electroporation, has been shown to facilitate naked DNA gene transfer in skeletal muscle allowing proteins to be produced and secreted at therapeutically relevant levels.
Plasmid DNA, unformulated or formulated with the non-ionic carrier SP1017, was injected at the rat tibialis anterior muscle followed by the application of electric pulses. Follow-up of protein expression was measured.
In our study we report that the non-ionic carrier SP1017 significantly increases transgene expression in rat muscle after the i.m. injection of a formulated-pCMVbeta plasmid followed by electroporation. Such increased expression was observed after delivering square-wave unipolar electric pulses at two different field strengths: low (110 V/cm) and high (175 V/cm). Moreover, elevated secreted placental alkaline phosphatase (SEAP) plasma levels were achieved with low-voltage (110 V/cm) electroporation. Our results also show that human hepatocyte growth factor (hHGF) can be produced from rat muscle and delivered to blood circulation at a biologically active level after a single i.m. injection of an SP1017-formulated plasmid (pCMV/hHGF) followed by electroporation. Tissue distribution studies mostly identified hHGF in the liver, but it was also found in the kidneys and lungs suggesting that here too the HGF could be of therapeutic benefit.
Our results indicate that SP1017 enhances the expression of electrotransferred genes. Such a delivery approach could prove an efficient method for the systemic production of therapeutic proteins.
治疗性蛋白质的全身给药是治疗多种疾病的常用方法。肌肉注射质粒后进行电穿孔，已被证明有助于裸DNA基因在骨骼肌中的转移，使蛋白质能够以治疗相关水平产生和分泌。
将未配制或与非离子载体SP1017配制的质粒DNA注射到大鼠胫前肌，随后施加电脉冲。测量蛋白质表达的随访情况。
在我们的研究中，我们报告在肌肉注射配制的pCMVβ质粒后进行电穿孔，非离子载体SP1017显著增加大鼠肌肉中的转基因表达。在以两种不同场强（低场强110V/cm和高场强175V/cm）施加方波单极电脉冲后观察到这种表达增加。此外，通过低电压（110V/cm）电穿孔实现了分泌型胎盘碱性磷酸酶（SEAP）血浆水平的升高。我们的结果还表明，单次肌肉注射SP1017配制的质粒（pCMV/hHGF）后进行电穿孔，大鼠肌肉可以产生人肝细胞生长因子（hHGF）并以生物活性水平输送到血液循环中。组织分布研究大多在肝脏中鉴定出hHGF，但在肾脏和肺中也发现了它，这表明在这里HGF也可能具有治疗益处。
我们的结果表明，SP1017增强了电转移基因的表达。这种递送方法可能证明是一种全身生产治疗性蛋白质的有效方法。
