Development of the parathyroid hormone production controlling method by the RNA interference in consideration of clinical application

考虑临床应用的RNA干扰甲状旁腺激素产生控制方法的开发

• 批准号: 22590916
• 负责人: TANAKA Reika
• 金额: $ 3.49万
• 依托单位: Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010-10-20 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22590916/
• 关键词: 副甲状腺機能亢進症; 二次性副甲状腺機能亢進症; ＲＮＡ干渉; RNA干渉; 副甲状腺ホルモン

Considering the safety to clinical application of gene therapy for secondary hyperparathyroidism (2HPT), we attempted the establishment of the RNA interference system in order to prevent damaging patient's genomes. Expression of miRNA for human parathyroid hormone (hPTH) was controlled with tetracycline inducible promotor in modified HEK293 cells which express hPTH constitutively. The cells were transplanted into nudemice, and secretion of hPTH into serum was confirmed. As the decrease of hPTH concentration in serum by feeding doxycycline was shown, the validity of this system was checked.In parallel to this, the survey of new target genes applied to this system found c-myc and p27Kip1, genes related with apoptosis of parathyroid cells.

考虑到基因治疗在继发性甲状旁腺功能亢进症（2HPT）中的临床应用安全性，我们尝试建立RNA干扰系统，以防止损害患者的基因组受损。在修饰的HEK293细胞中，用四环素可诱导型启动子控制人类甲状旁腺激素（HPTH）miRNA的表达，这些启动子在组成性地表达HPTH。将细胞移植到裸性中，并确认HPTH分泌到血清中。由于显示了多西环素在血清中HPTH浓度的降低，因此检查了该系统的有效性。在此同时，对应用于该系统的新靶基因的调查发现了与甲状旁腺细胞凋亡相关的C-MYC和P27KIP1。