Transfection of NFκB-decoy oligodeoxynucleotides using efficient ultrasound-mediated gene transfer into donor kidneys prolonged survival of rat renal allografts
Transfection of NFκB-decoy oligodeoxynucleotides using efficient ultrasound-mediated gene transfer into donor kidneys prolonged survival of rat renal allografts
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DOI：
10.1038/sj.gt.3301882
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发表时间：
2003-03-01
期刊：
影响因子：
5.1
通讯作者：
Morishita, R
中科院分区：
文献类型：
--
作者：
Azuma, H;Tomita, N;Morishita, R
核因子-κ B（NF-κ B）在一系列细胞因子和粘附分子基因的协同反式激活中起着关键作用，这些基因与器官移植急性排斥反应的发生密切相关。我们先前开发了针对NF κ B的诱饵顺式元件寡脱氧核糖核酸（NF κ B-decoy），其在体外和体内有效地抑制主要炎症介质的活化。因此，我们假设将NF κ B-诱饵转染到供体肾中可以防止急性排斥反应并延长移植物存活，从而为肾急性排斥反应提供有效的治疗。为了检测NF κ B-诱饵，我们采用了一种新的方法，使用超声暴露与超声心动图造影剂，Optison，并清楚地表明成功转染NF κ B-诱饵到肾组织。然后在大鼠肾同种异体移植物模型（Wistar-Lewis）中评价NF κ B-诱饵对肾同种异体移植物的治疗效果。在对照组中，移植物功能显著恶化，肾组织明显破坏，伴随着主要炎症介质的产生增加，所有动物在9天内死于肾衰竭。相比之下，移植物功能（第2天血清肌酐，NF κ B治疗。0.97+/- 0.16 vs对照组：1.84 +/- 0.23 mg/dl，P < 0.01），组织学结构保存良好，NF κ B调节的细胞因子和粘附分子的表达显著降低，包括IL-1、iNOS、MCP-1、TNF-α和ICAM-1。结果，与对照组相比，该组动物的存活时间显著延长（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，并证明了移植物存活的显着延长NF κ B-诱饵成功转染到大鼠肾移植模型中的供体肾。
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.