Azuma H, Tomita N, Kaneda Y, Koike H, Ogihara T, Katsuoka Y, Morishita R
Department of Urology, Osaka Medical College, Takatsuki, Osaka, Japan.
Gene Ther. 2003 Mar;10(5):415-25. doi: 10.1038/sj.gt.3301882.
Nuclear factor kappaB (NFkappaB) plays a pivotal role in the coordinated transactivation of a series of genes of cytokines and adhesion molecules that are highly involved in the onset of acute rejection in organ transplantation. We previously developed decoy cis-elements oligo deoxyribonucleic acid against NFkappaB (NFkappaB-decoy) that effectively inhibited the activation of major inflammatory mediators in vitro and in vivo. Accordingly, we hypothesized that transfection of NFkappaB-decoy into the donor kidney would prevent acute rejection and prolong graft survival, and thus provide effective therapy for renal acute rejection. To transfect NFkappaB-decoy, we employed a novel approach using ultrasound exposure with an echocardiographic contrast agent, Optison, and clearly demonstrated successful transfection of NFkappaB-decoy into renal tissue. The therapeutic effect of NFkappaB-decoy on renal allografts was then evaluated in a rat renal allograft model (Wistar-Lewis). In the control group, graft function significantly deteriorated with marked destruction of renal tissue, accompanied by increased production of major inflammatory mediators, and all animals died of renal failure by 9 days. In contrast, graft function (serum creatinine on day 2, NFkappaB-treated: 0.97+/-0.16 versus control: 1.84+/-0.23 mg/dl, P<0.01) and histological structure were well preserved with significantly decreased expression of NFkappaB-regulated cytokines and adhesion molecules, including IL-1, iNOS, MCP-1, TNF-alpha, and ICAM-1, in allografts transfected with NFkappaB-decoy. As a result, animal survival was significantly prolonged in this group as compared to controls (14.2+/-5.2 versus 7.1+/-1.2 days, P<0.01). Thus, we established a novel ultrasound-Optison-mediated gene transfection approach and demonstrated the significant prolongation of graft survival by the successful transfection of NFkappaB-decoy into the donor kidney in a rat renal allograft model.
核因子κB（NFκB）在一系列细胞因子和黏附分子基因的协同反式激活中起关键作用，这些基因与器官移植急性排斥反应的发生密切相关。我们之前开发了针对NFκB的诱饵顺式元件寡脱氧核糖核酸（NFκB诱饵），其在体外和体内均能有效抑制主要炎症介质的激活。因此，我们推测将NFκB诱饵转染到供体肾中可预防急性排斥反应并延长移植物存活时间，从而为肾急性排斥反应提供有效的治疗方法。为了转染NFκB诱饵，我们采用了一种使用超声造影剂Optison进行超声暴露的新方法，并清楚地证明了NFκB诱饵成功转染到肾组织中。然后在大鼠肾移植模型（Wistar-Lewis）中评估NFκB诱饵对肾移植的治疗效果。在对照组中，移植物功能显著恶化，肾组织明显破坏，同时主要炎症介质的产生增加，所有动物均在9天内死于肾衰竭。相比之下，在用NFκB诱饵转染的移植肾中，移植物功能（第2天血清肌酐，NFκB处理组：0.97±0.16，对照组：1.84±0.23mg/dl，P<0.01）和组织结构得到良好保存，NFκB调节的细胞因子和黏附分子，包括IL-1、iNOS、MCP-1、TNF-α和ICAM-1的表达显著降低。结果，与对照组相比，该组动物存活时间显著延长（14.2±5.2天对7.1±1.2天，P<0.01）。因此，我们建立了一种新的超声-Optison介导的基因转染方法，并在大鼠肾移植模型中证明了通过将NFκB诱饵成功转染到供体肾中可显著延长移植物存活时间。
