Reseach for Gene therapy of ARJP

ARJP基因治疗研究

• 批准号: 11470150
• 负责人: MOCHIZUKI Hideki
• 金额: $ 1.79万
• 依托单位: JUNTENDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470150/
• 关键词: ARJP; Parkinson's disease; AAV vector; Parkin gene; Gene therapy; 常染色体劣性遺伝若年性パーキンソン病

In the studies of familiar parkinsonism, the parkin protein has recently been clarified from the analysis of autosomal recessive juvenile parkinsonism (ARJP). ARJP is characterized by selective dopaminergic cell death and the absence of Lewy bodies. Mutations of the parkin gene cause ARJP.Parkin protein is involved in protein degradation as an ubiquitin-protein ligase collaborating with the ubiquitin-conjugating enzyme UbcH7. A mutant of parkin protein from ARJP patients showed loss of its ubiquitin-protein ligase activity.Because ARJP involves loss of function, gene therapy using the parkin gene may prevent nigral cell death as a form of anti-apoptotic therapy. Therefore, we established AAV vector system to express Parkin protein for gene therapy in ARJP patients.The advantages of AAV vector are that it is non-pathogenic, it has stable integration and has the ability to introduce a gene into non-dividing cells. The titer is 108 cfu/ml. Only 5 ul of AAV-GFP was stereotactically injected into the striatum. We got high expression of Parkin protein in the normal mouse striatum. We are going to make knock out mouse for the replacement gene therapy experiment of ARJP.

在常见的帕金森综合征的研究中，Parkin蛋白最近从常染色体隐性遗传性青少年帕金森综合征(ARJP)的分析中得到澄清。ARJP的特点是选择性多巴胺能细胞死亡和路易小体缺失。Parkin基因的突变导致ARJP。Parkin蛋白作为泛素蛋白连接酶与泛素结合酶UbcH7合作参与蛋白质降解。ARJP患者的parkin蛋白突变体显示其泛素蛋白连接酶活性丧失，由于ARJP涉及功能丧失，使用parkin基因进行基因治疗可能会防止黑质细胞死亡，作为抗凋亡治疗的一种形式。因此，我们建立了表达Parkin蛋白的AAV载体系统，用于ARJP患者的基因治疗，其优点是无致病性，具有稳定的整合性，并具有将基因导入非分裂细胞的能力。AAV-GFP的滴度为108CFU/ml，仅5ul的AAV-GFP立体定向注射到纹状体。Parkin蛋白在正常小鼠纹状体中高表达。我们将为ARJP的替代基因治疗实验制作基因敲除小鼠。

项目成果

Mochizuki H: "Pathogenesis and neuronal death ; Parkinson's disease"Clinical Neuroscience. 18. 84-86 (2000)

Mochizuki H：“发病机制和神经元死亡；帕金森病”临床神经科学。

Nakano K,Migita M,Mochizuki H. et al.: "Differentiation of transplanted bone marrow cells in the adult mouse brain."Transplantation. (In press). (2000)

Nakano K、Migita M、Mochizuki H. 等人：“成年小鼠大脑中移植骨髓细胞的分化。”移植。

望月秀樹: "病態と神経細胞死:パーキンソン病"Clinaical Neuroscience. 18. 84-86 (2000)

Hideki Mochizuki：“病理学和神经元死亡：帕金森病”临床神经科学 18. 84-86 (2000)。

望月秀樹,後藤啓五,水野美邦: "パーキンソン病の分子生物学"感染 炎症 免疫. 30-4. 12-19 (2000)

Hideki Mochizuki、Keigo Goto、Mikuni Mizuno：“帕金森病的分子生物学”感染炎症免疫学 12-4。

Takanashi M,Mochizuki H,Yokomizo K. et al.: "Iron accumulation in the substantia nigra of ARJP."Parkinsonism and related disorders. (In press). (2000)

Takanashi M、Mochizuki H、Yokomizo K. 等人：“ARJP 黑质中的铁积累。”帕金森症和相关疾病。