Bloquel Carole, Bessis Natacha, Boissier Marie-Christophe, Scherman Daniel, Bigey Pascal
UPRES EA-3408 and Service de Rhumatologie, UFR Léonard de Vinci, Université Paris 13 and Hopital Avicenne (AP-HP), 93017 Bobigny Cedex, France.
Hum Gene Ther. 2004 Feb;15(2):189-201. doi: 10.1089/104303404772679995.
Electrotransfer is a simple and efficient strategy of nonviral gene delivery. We have used this method to deliver plasmids encoding three human tumor necrosis factor-alpha soluble receptor I variants (hTNFR-Is) a monomeric hTNFR-Is, a chimeric hTNFR-Is/mIgG1, and a dimeric (hTNFR-Is)(2) form. Electrotransfer parameters were studied and because anti-TNF strategies have proven efficient for the treatment of rheumatoid arthritis in clinics, we used a collagen-induced arthritis (CIA) mouse model to assess the efficacy of our constructs in the treatment of the disease. All proteins were proven bioactive, both in vitro and ex vivo. Plasmid intramuscular electrotransfer in mice resulted in a local expression of the three variants for at least 6 months; systemic expression lasted also more than 6 months for the hTNFR-Is/mIgG1 form, while it was shorter for the two other forms. This expression was plasmid dose-dependent. Electrotransfer of 50 microg of hTNFR-Is/mIgG1 at the onset of a CIA induced a clear-cut decrease in both clinical and histologic signs of the disease; the dimeric form also showed some efficacy. Moreover, the long-lasting protective effect was observed for more than 5 weeks. Comparison of this electrotransfer approach with repeated recombinant protein (etanercept) injections highlighted the potential practical interest of gene therapy approach for CIA, which leads to sustained therapeutic effect after single treatment. These results show that electrotransfer may be a useful method to deliver cytokine or anticytokine therapy in rheumatoid arthritis and also illustrate the potentiality of plasmid intramuscular electrotransfer for the rapid screening and assessment of different variant forms of secreted proteins.
电穿孔转移是一种简单有效的非病毒基因递送策略。我们已使用该方法递送编码三种人肿瘤坏死因子-α可溶性受体I变体（hTNFR-Is）的质粒，即单体hTNFR-Is、嵌合型hTNFR-Is/mIgG1和二聚体（hTNFR-Is）2形式。我们研究了电穿孔转移参数，并且由于抗TNF策略已在临床上被证明对类风湿性关节炎的治疗有效，因此我们使用胶原诱导的关节炎（CIA）小鼠模型来评估我们构建体对该疾病的治疗效果。所有蛋白质在体外和体内均被证明具有生物活性。小鼠体内质粒肌肉内电穿孔转移导致三种变体在局部表达至少6个月；hTNFR-Is/mIgG1形式的全身表达也持续超过6个月，而其他两种形式的全身表达持续时间较短。这种表达呈质粒剂量依赖性。在CIA发病时电穿孔转移50μg的hTNFR-Is/mIgG1可使疾病的临床和组织学症状明显减轻；二聚体形式也显示出一定疗效。此外，观察到了超过5周的持久保护作用。将这种电穿孔转移方法与重复注射重组蛋白（依那西普）进行比较，突出了基因治疗方法对CIA的潜在实际应用价值，该方法在单次治疗后可产生持续的治疗效果。这些结果表明，电穿孔转移可能是一种在类风湿性关节炎中递送细胞因子或抗细胞因子疗法的有用方法，也说明了质粒肌肉内电穿孔转移在快速筛选和评估分泌蛋白不同变体形式方面的潜力。
