Adriaansen J, Khoury M, de Cortie C J, Fallaux F J, Bigey P, Scherman D, Gould D J, Chernajovsky Y, Apparailly F, Jorgensen C, Vervoordeldonk M J B M, Tak P P
Division of Clinical Immunology and Rheumatology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
Ann Rheum Dis. 2007 Sep;66(9):1143-50. doi: 10.1136/ard.2006.064519. Epub 2007 Mar 15.
In the context of preclinical development, we studied the potential of intra-articular gene delivery using a recombinant adeno-associated virus 5 (rAAV5) encoding a chimeric human tumour necrosis factoralpha (TNFalpha) soluble receptor I linked to a mouse immunoglobulin heavy chain Fc portion (TNF receptor I; TNFRI-Ig).
Expression was under control of a nuclear factor kappa B (NFkappaB)-responsive promoter and compared with a cytomegalovirus (CMV) promoter (rAAV5.NFkappaB-TNFRI-Ig and rAAV5.CMV-TNFRI-Ig, respectively).
Fibroblast-like synoviocytes transduced in vitro with rAAV5.NFkappaB-TNFRI-Ig were able to produce TNFRI-Ig protein in response to several stimuli, and this was inhibited upon treatment with a specific NFkappaB blocking agent. A bioassay revealed that the synthesised TNFRI-Ig was bioactive, showing a higher affinity for human than for rat TNFalpha. Transcription of the transgene and protein production were detectable in joints injected with both constructs. No dissemination of the vector was observed outside the joints. A significant reduction in paw swelling was seen in rats treated with rAAV5.NFkappaB-TNFRI-Ig. This clinical effect was accompanied by a decrease in pro-inflammatory cytokine levels and an increase in IL10 expression in the synovium.
These results provide evidence that intra-articular gene therapy using rAAV5 encoding TNFRI-Ig may be a safe and feasible approach for the treatment of rheumatoid arthritis. The higher affinity for human TNFalpha suggests that in patients with rheumatoid arthritis the therapeutic effect might be even more pronounced than in rat adjuvant arthritis.
在临床前开发的背景下，我们研究了使用编码与小鼠免疫球蛋白重链Fc部分相连的嵌合型人肿瘤坏死因子α（TNFα）可溶性受体I的重组腺相关病毒5（rAAV5）进行关节内基因递送的潜力。
表达受核因子κB（NFκB）反应性启动子控制，并与巨细胞病毒（CMV）启动子进行比较（分别为rAAV5.NFκB-TNFRI-Ig和rAAV5.CMV-TNFRI-Ig）。
用rAAV5.NFκB-TNFRI-Ig体外转导的成纤维样滑膜细胞能够在受到多种刺激时产生TNFRI-Ig蛋白，并且在用特异性NFκB阻断剂处理后这种产生受到抑制。生物测定显示合成的TNFRI-Ig具有生物活性，对人TNFα的亲和力高于对大鼠TNFα的亲和力。在注射了两种构建体的关节中均可检测到转基因的转录和蛋白产生。未观察到载体在关节外扩散。用rAAV5.NFκB-TNFRI-Ig处理的大鼠爪肿胀明显减轻。这种临床效果伴随着促炎细胞因子水平的降低和滑膜中IL10表达的增加。
这些结果提供了证据，表明使用编码TNFRI-Ig的rAAV5进行关节内基因治疗可能是治疗类风湿性关节炎的一种安全可行的方法。对人TNFα的较高亲和力表明，在类风湿性关节炎患者中，治疗效果可能比对大鼠佐剂性关节炎更明显。
