Inhibition of neuronal cell death in Parkinson's disease by transduction of neurotrophic factor gene.

通过神经营养因子基因转导抑制帕金森病中的神经细胞死亡。

• 批准号: 11670640
• 负责人: MORI Hideo
• 金额: $ 1.34万
• 依托单位: JUNTENDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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/grant/KAKENHI-PROJECT-11670640/
• 关键词: Pigment epitheliurm-derived factori; Neurotrophic factor; Substantia nigra; Striatum; Apoptosis; Adeno-associated virus; Gene therapy; PEDF; 神経栄養因子; 黒質神経細胞死; パーキンソン病; レトロウィルスベクター; アストロサイト; マイクログリア; 移植; 黒質神経細胞

Pigment epithelium-derived factor (PEDF), a neurotrophic agents first identified in conditioned medium from cultured human retinal pigment epithelial cells, induces neuronal differentiation with neuritic outgrowth in Y-79 retinoblastoma cells and has a neurotrophic survival effect on cerebellar granule cells in culture. PEDF also have protective effect against glutamate toxicity and induction of apoptotic cell death of cultured granular cells. In this project, first we investigated the effects of human recombinant PEDF (hrPEDF) on proliferation of cultured nigral neurones. To do this experiments, we used cultures of nigral neurones isolated from the fetal rat brains. The number of nigral neurones, which identified by immunohistochemistry for tyrosine hydroxilase, is larger in the presence of PEDF than that of controls. The effect of PEDF showed a dose-response relationship. In second studies, we have performed the transduction of PEDF gene to the striatum and nigra of mice. To this purpose, we used recombinent adeno-associated (AAV) vectors which are sufficient for packaging and integration. AAV containing cDNA encoding PEDF and GFP were injected to the striatum of the mice by streotaxic operation. GFP-positive cells were observed in the striatum and the substantia nigra, which were transported through strionigral pathway. We are now conducting immunohistochemistry for PEDF on these sections to confirm the expression of the PEDF.Our study suggest the possibility of PEDF gene transfer to the striatum and nigra and effect of survival of niral neurones, and furthermore potential role for the gene therapy of Parkinson's disease

色素上皮衍生因子（PEDF）是一种神经营养因子，在培养的人视网膜色素上皮细胞的条件培养液中首次发现，它能诱导Y-79视网膜母细胞瘤细胞向神经元分化并产生神经炎样生长，对培养的小脑颗粒细胞具有神经营养存活作用。PEDF对谷氨酸毒性和诱导颗粒细胞凋亡也有保护作用。在本研究中，我们首先研究了人重组PEDF（hrPEDF）对培养的黑质神经元增殖的影响。为了做这个实验，我们使用了从胎鼠大脑中分离的黑质神经元的培养物。酪氨酸羟化酶免疫组化结果显示，PEDF组黑质神经元的数量明显多于对照组。PEDF的作用呈剂量-反应关系。在第二个研究中，我们进行了PEDF基因转导到小鼠的纹状体和黑质。为此目的，我们使用重组腺相关（AAV）载体，其足以进行包装和整合。将含有编码PEDF和GFP的cDNA的AAV注射到小鼠的纹状体中。在纹状体和黑质中观察到GFP阳性细胞，其通过纹状体黑质途径转运。本研究旨在探讨PEDF基因转移到纹状体和黑质的可能性及其对神经元存活的影响，并为帕金森病的基因治疗奠定基础

项目成果

Mizuno Y, Hattori N, Mori H: "Genetics of Parkinson's disease."Biomed Pharmacother. 53. 109-116 (1999)

Mizuno Y、Hattori N、Mori H：“帕金森病的遗传学”。Biomed Pharmacother。

Mizuno Y,Hattori N,Mori H: "Genetics of Parkinson's desease"Biomed Phormacother. 59. 109-116 (1999)

Mizuno Y，Hattori N，Mori H：“帕金森病的遗传学”Biomed Phormacother。

Sugita Y, Zhao B, Shankar P, Dunbar CE, Doren S, Young HA, Schwartz JP: "CNS interleukin-3 (IL-3) expression and neurological syndrome in antisense-IL-3 transgenic mice."J Neuropathol Exp Neurol. 58. 480-8 (1999)

Sugita Y、Zhao B、Shankar P、Dunbar CE、Doren S、Young HA、Schwartz JP：“反义 IL-3 转基因小鼠中的中枢神经系统白细胞介素 3 (IL-3) 表达和神经系统综合征。”J Neuropathol Exp Neurol。

Sugita Y,Zhao B,Shankar P,Dunbar CE,Doren S,Young HA,Schwartz JP: "CNS interleukin-3(IL-3) expression and neurological syndrome in antisense-IL-3 transgenic mice."J Neuropathol Exp Neurol. 58. 480-488 (1999)

Sugita Y、Zhao B、Shankar P、Dunbar CE、Doren S、Young HA、Schwartz JP：“反义 IL-3 转基因小鼠中枢神经系统白细胞介素 3 (IL-3) 的表达和神经系统综合征。”J Neuropathol Exp Neurol。

Takanashi M,Mochizuki H,Yokomizo K,Hattori N,Mori H,et al.: "Iron accumulation in the substantia nigra of autosomal recessive javenile parkinsonism"Parkinson's disease and related disorders. in press.

Takanashi M，Mochizuki H，Yokomizo K，Hattori N，Mori H，et al.：“常染色体隐性青少年帕金森病黑质中的铁积累”帕金森病和相关疾病。