权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development of gene therapy for progressive renal diseases by ribozyme

核酶治疗进行性肾病的基因疗法的发展

基本信息

批准号：
15590863
负责人：
FUKUDA Noboru
金额：
$ 2.24万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590863/
关键词：
PDGF A-chain TGF-b1 ribozyme adeno virus kidney disease TGF-β1

项目摘要

To develop ribozyme therapy for progressive renal diseases, we designed and synthesized the nucleic acid resistant DNA/RNA chimeric ribozyme targeting PDGF A-chain and TGF-b1 by DNA/RNA synthesizer. In addition, we also obtained the recombinant ribozyme targeting PDGF A-chain and TGF-b1 adeno virus vector. FITC-labeled DNA/RNA chimeric ribozymes were considerably delivered into cultured mesangial cells with zelatin delivery reagent. The ribozymes significantly inhibited expression of PDGF A-chain and TGF-b1 mRNA in mesangial cells in vitro.Based on the in vitro experimental results, we investigated in vivo delivery and effects of the DNA/RNA chimeric ribozyme targeting PDGF A-chain and TGF-b1 on progressive renel diseases in Dahl-salt sensitive rats. FITC-labeled DNA/RNA chimeric ribozymes injected intravenously were sufficinatly delivered into gromeruli and nephrotubulus in Dahl-salt sensitive rats. The DNA/RNA chimeric ribozymes with the zelatin delivery reagent and the recombinant ribozymes significantly inhibited expressions of PDGF A-chain and TGF-b1 mRNAs and proteins in renal cortex, and decreased urinary protein excressions to half levels. Intra venous injections of excess amount of the DNA/RNA chimeric ribozymes with the zelatin delivery reagent and the recombinant ribozymes did not induce tissue damages in spleen, bone marrow, testis, kideney and heart, indicating that safety of the ribozyme treatments in vivo.These results suggest that the the DNA/RNA chimeric ribozymes with the zelatin delivery reagent and the recombinant ribozymes targeting PDGF A-chain and TGF-b1 will be feasible gene therapy for the progressive renal diseases.

为开发核酶治疗进展性肾脏疾病的新方法，我们设计并合成了靶向PDGF A链和TGF-β 1的核酸抗性DNA/RNA嵌合核酶。此外，我们还获得了靶向PDGF A链的重组核酶和TGF-β 1腺病毒载体。FITC标记的DNA/RNA嵌合核酶被大量地递送到培养的系膜细胞中。在体外实验的基础上，我们研究了靶向PDGF A链和TGF-β 1的DNA/RNA嵌合核酶的体内给药及其对Dahl盐敏感大鼠肾脏疾病的治疗作用。经静脉注射FITC标记的DNA/RNA嵌合核酶可有效地进入Dahl盐敏感大鼠的肾小球和肾小管。DNA/RNA嵌合核酶与zelatin载体及重组核酶均能显著抑制肾皮质PDGF A链和TGF-β 1 mRNA和蛋白的表达，并使尿蛋白排泄量减半。静脉内注射过量的DNA/RNA嵌合核酶与zelatin递送试剂和重组核酶不诱导脾、骨髓、睾丸、肾脏和心脏中的组织损伤，这些结果表明，具有zelatin递送试剂的DNA/RNA嵌合核酶和靶向PDGF A链和TGF-β的重组核酶在体内是安全的。b1基因治疗进展性肾脏疾病是可行的。