Richard Peggy, Pollard Hélène, Lanctin Caroline, Bello-Roufaï Mahajoub, Désigaux Léa, Escande Denis, Pitard Bruno
L'Institut du Thorax, Institut National de la Santé et de la Recherche Médicale, Inserm U533, Faculté de Médecine, 44000 Nantes, France.
J Gene Med. 2005 Jan;7(1):80-6. doi: 10.1002/jgm.631.
We have previously shown that intramuscular injection of plasmid DNA formulated with a non-ionic amphiphile synthetic vector [poly(ethylene oxide)(13)-poly(propylene oxide)(30)-poly(ethylene oxide)(13) block copolymer; PE6400] increases reporter gene expression compared with naked DNA. We have now investigated this simple non-viral formulation for production of secreted proteins from the mouse skeletal muscle.
Plasmids encoding either constitutive human secreted alkaline phosphatase or murine erythropoietin inducible via a Tet-on system were formulated with PE6400 and intramuscularly injected into the mouse tibial anterior muscle.
PE6400/DNA formulation led to an increased amount of recombinant alkaline phosphatase secreted from skeletal muscle as compared with naked DNA. In the presence of doxycycline, a single injection of 10 microg plasmid encoding inducible murine erythropoietin formulated with PE6400 significantly increased the hematocrit, whereas the same amount of DNA in the absence of PE6400 had no effect. The increase in the hematocrit was stable for 42 days. The tetracycline-inducible promoter permitted pharmacological control of hematocrit level after DNA intramuscular injection. However, 4 months post-injection the hematocrit returned to its pre-injection value, even in the presence of doxycycline. This phenomenon was likely caused by an immune response against the tetracycline-activated transcription factor.
Intramuscular injection of plasmid DNA formulated with PE6400 provides an efficient and simple method for secretion and production of non-muscle proteins.
我们之前已经表明，与裸DNA相比，肌肉注射用非离子两亲性合成载体[聚（环氧乙烷）（13）-聚（环氧丙烷）（30）-聚（环氧乙烷）（13）嵌段共聚物；PE6400]配制的质粒DNA可增加报告基因的表达。我们现在已经研究了这种简单的非病毒制剂用于从小鼠骨骼肌中产生分泌蛋白。
将编码组成型人分泌碱性磷酸酶或通过Tet-on系统诱导的小鼠促红细胞生成素的质粒与PE6400配制，并肌肉注射到小鼠胫前肌中。
与裸DNA相比，PE6400/DNA制剂导致骨骼肌分泌的重组碱性磷酸酶量增加。在强力霉素存在下，单次注射10微克用PE6400配制的编码诱导型小鼠促红细胞生成素的质粒可显著提高血细胞比容，而在没有PE6400的情况下相同量的DNA则没有效果。血细胞比容的增加在42天内保持稳定。四环素诱导型启动子允许在DNA肌肉注射后对血细胞比容水平进行药理学控制。然而，注射后4个月，即使在强力霉素存在下，血细胞比容也恢复到注射前的值。这种现象可能是由针对四环素激活转录因子的免疫反应引起的。
肌肉注射用PE6400配制的质粒DNA为非肌肉蛋白的分泌和产生提供了一种有效且简单的方法。
