Development of gene silencing pyrrole-imidazole polyamide targeting the TGF-β1 promoter for treatment of progressive renal diseases
Development of gene silencing pyrrole-imidazole polyamide targeting the TGF-β1 promoter for treatment of progressive renal diseases
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DOI：
10.1681/asn.2005060650
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发表时间：
2006-02-01
影响因子：
13.6
通讯作者：
Serie, Kazuo
中科院分区：
文献类型：
--
作者：
Fukuda, Noboru;Ueno, Takahiro;Serie, Kazuo
吡咯-咪唑（Py-Im）聚酰胺是一种新型的抗核酸酶化合物，通过与DNA小沟结合来抑制基因表达。一种靶向大鼠TGF-β 1的Py-Im聚酰胺被设计为用于进行性肾脏疾病的基因沉默剂，并在达尔盐敏感（Dahl-S）大鼠中检查该聚酰胺的分布和对肾损伤的影响。为了鉴定用于激活大鼠TGF-β 1基因的转录因子结合元件，将重组TGF-β 1报告质粒转染到HEK-293细胞中，并测量启动子活性。Py-Im聚酰胺被设计为大鼠TGF-β 1启动子的激活蛋白-1结合位点。该Py-Im聚酰胺在凝胶迁移率变化和Biacore测定中显示出与靶DNA的强、快速和特异性结合。Py-Im聚酰胺显著抑制大鼠系膜细胞TGF-β 1启动子活性及TGF-β 1 mRNA和蛋白表达。腹腔注射荧光素标记的聚酰胺分布到大鼠的肾脏。Py-Im聚酰胺显著抑制Dahl-S大鼠肾皮质中TGF-β 1 mRNA和蛋白的表达，并降低Dahl-S大鼠尿蛋白和白蛋白的增加，而与血压的变化无关。这些结果表明，靶向TGF-β 1的Py-Im聚酰胺将成为TGF-β 1相关疾病（包括进行性肾病）的新型基因沉默剂。
Pyrrole-imidazole (Py-Im) polyamides are nuclease-resistant novel compounds that inhibit gene expression by binding to the minor groove of DNA. A Py-Im polyamide that targets rat TGF-beta 1 was designed as a gene-silencing agent for progressive renal diseases, and the distribution and the effects of this polyamide on renal injury were examined in Dahl-salt sensitive (Dahl-S) rats. For identification of transcription factor binding elements for activation of the rat TGF-beta 1 gene, recombinant TGF-beta 1 reporter plasmids were transfected into HEK-293 cells, and promoter activity was measured. Py-Im polyamide was designed to the activator protein-1 binding site of the rat TGF-beta 1 promoter. This Py-Im polyamide showed strong, fast, and specific binding to the target DNA in gel mobility shift and Biacore assays. Py-Im polyamide significantly inhibited TGF-beta 1 promoter activity and expression of TGF-beta 1 mRNA and protein in rat mesangial cells. Intravenously administered fluorescein-labeled polyamide distributed to the kidney of rats. Py-Im polyamide significantly inhibited expression of TGF-beta 1 mRNA and protein in the renal cortex of Dahl-S rats and reduced the increase in urinary protein and albumin in Dahl-S rats independent of changes in blood pressure. These results indicate that Py-Im polyamide that targets TGF-beta 1 will be a novel gene-silencing agent for the TGF-beta 1-associated diseases, including progressive renal diseases.