Basic studies for an effective therapy for Alport syndrome

阿尔波特综合征有效治疗的基础研究

• 批准号: 13470159
• 负责人: UCHIYAMA Makoto
• 金额: $ 5.25万
• 依托单位: NIIGATA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470159/
• 关键词: Alport syndrome; cyclosporin A; enalapril; tenascin C; Col4a4; gene therapy; electroporation; trancegenic mouse; サイクロポリンA; ミゾリビン

1. Study for the efficiency of drug therapy for Alport syndrome using Col4a4 deficient miceCompared with untreated and mizoribine-treated groups, proteinuria and crescent formation were significantly reduced in cyclosporine A-treated and enalapril-treated groups. Furthermore, enalapril could significantly prolong the survival period of mutant mice, although cyclosporine A and mizoribine could not. Taken together, angiotensin converting enzyme inhibitor was thought to be effective in slowing down the progression towards renal failure in a mouse model of Alport syndrome.2. Analysis of Col4a4, tenascin double deficient miceCompared with Col4a4 deficient mice, proteinuria and survival period were significantly improved in Col4a4, tenascin double deficient mice. Elavation of tenascin gene expression observed at the glomerular basement membrane of Col4a4 deficient mice was thought to play a role for promote the pathological change of Alport syndrome.3. Rescue study of the phenotype of Col4a4 deficient mice by Col4a4 transgenic miceCol4a4 transgenic mice carrying CAG promoter and LacZ reporter gene were produced. The mice were mated with Col4a4 deficient mice. We are now examining whether the phenotype of Col4a4 deficient mice is rescued or not.4. Study of gene therapy for Alport syndrome using electroporationThe plasmid pCAG- Col4a4-LacZ was electroporated into mouse kidneys by either abdominal aorta route or renal capsule route. Although efficiency of DNA introduction via abdominal aorta was very low, considerable number of cells in renal cortex were introduced the plasmid by electroporation from subcapsular route. The electroporation from subcapsular route may be available for gene therapy for Alport syndrome and other renal diseases.

1. Col4a4缺陷小鼠治疗Alport综合征的疗效研究与未治疗组和米佐利比组相比，环孢素a治疗组和依那普利治疗组的蛋白尿和新月形成明显减少。此外，依那普利能显著延长突变小鼠的生存期，而环孢素A和米佐利滨则不能。综上所述，血管紧张素转换酶抑制剂被认为对减缓Alport综合征小鼠模型肾功能衰竭的进展是有效的。Col4a4， tenascin双缺陷小鼠与Col4a4缺陷小鼠相比，Col4a4， tenascin双缺陷小鼠的蛋白尿和生存期明显改善。Col4a4缺陷小鼠肾小球基底膜tenascin基因表达升高被认为促进了Alport综合征的病理改变。利用Col4a4转基因小鼠对Col4a4缺陷小鼠表型的拯救研究制备了携带CAG启动子和LacZ报告基因的Col4a4转基因小鼠。这些小鼠与Col4a4缺陷小鼠交配。我们现在正在研究Col4a4缺陷小鼠的表型是否被拯救。电穿孔基因治疗Alport综合征的研究pCAG- Col4a4-LacZ质粒经腹主动脉或肾包膜途径电穿孔进入小鼠肾脏。虽然经腹主动脉导入DNA的效率很低，但仍有相当数量的肾皮质细胞通过荚膜下电穿孔途径导入质粒。荚膜下电穿孔可用于Alport综合征和其他肾脏疾病的基因治疗。