Therapy for neurodegeneerative disease using novel screening method for chemical chaperon.

使用化学伴侣的新型筛选方法治疗神经退行性疾病。

• 批准号: 18590967
• 负责人: OSAKA Hitoshi
• 金额: $ 2.04万
• 依托单位: Kanagawa Cancer Center Research Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590967/
• 关键词: Biological neurochemistry; Neurodegeneration; Chaperon; 小胞体ストレス

(1)Therapy for Pelizaeus-Merzbacher with chaperon. We failed to establish the gas-chromatographically measurement for a candidate chemical. We switched to search for chemicals to decrease the expression level of proteolipid protein(PLP). Duplication of PLP accounts for the half of the PMD patients. In C6 glioma cell line expressing PLP, we could measure PLP mRNA level by Quantitative RT-PCR. PLP expression was normalized by that of hypoxanthine guanine phosphoribosyl transferase. At first, we found that the more cell differentiate, the more PLP expresses. We optimized screening condition as follows ; chemicals extracted from food were added at final concentration of 10μM at cell density of 2.5×10^5/well(9.2cm^2) for 48hrs with DMEM(low glucose) including 1%fetal bovine serum. We screened the library including more than one hundred of food chemicals. We found 2 chemicals that decrease the PLP mRNA expression more than 50%. 11 chemicals were found to decrease the PLP expression level mor … More e than 30%. We have already found that PLP missence mutations cause overload to proteorytic system in the cell leading to cell death. Therefore, food chemicals that lower the level of PLP may be applied to the PMD patients with duplication as well as missence mutations in PLP gene.(2)Establishment of therapeutic effect in mouse model of PMD. For proper evaluation of drugs for PMD, we need to assess the degree of myelination in mouse model. We tried to quantitatively measure myelination by microscopic examinations. Firstly, we evaluated the degree of myelination using light microscopic examinations after conventional HE, Kluver-Barrera's or immune staining using the antibodies to PLP and myelin basic protein. These methods partially disclosed the dysmyelination of PMD model mouse. Electron microscopic examination revealed the degree of myelination properly. Optical nerve was superior for quantitative examination among central nervous system, as it contained the uniform numbers of neuron and oligodendrocytes. Less

（1）Pelizaeus-Merzbacher二氏病伴伴侣治疗。我们未能建立一个候选化学品的气相色谱测量。我们转而寻找化学物质来降低蛋白脂质蛋白（PLP）的表达水平。在PMD患者中，PLP重复占一半。在表达PLP的C6胶质瘤细胞系中，我们可以通过定量RT-PCR检测PLP mRNA的水平。PLP表达通过次黄嘌呤鸟嘌呤磷酸核糖转移酶的表达标准化。首先，我们发现细胞分化越多，PLP表达越多。筛选条件优化为：以含1%胎牛血清的DMEM（低糖）培养液为培养基，加入终浓度为10μM的食物提取物，细胞密度为2.5×10^5/孔（9.2cm ^2），培养48小时。我们筛选了包括一百多种食品化学品的库。我们发现2种化学物质使PLP mRNA表达降低50%以上。发现11种化学物质降低PLP表达水平莫尔 ...更多信息 超过30%。我们已经发现PLP缺失突变导致细胞中蛋白质降解系统过载，导致细胞死亡。因此，降低PLP水平的食品化学品可应用于PLP基因重复突变和错义突变的PMD患者。（2）PMD小鼠模型治疗效果的建立。为了正确评价PMD药物，我们需要评估小鼠模型中髓鞘形成的程度。我们试图通过显微镜检查定量测量髓鞘形成。首先，我们在常规HE、Kluver-Barrera’s或使用PLP和髓鞘碱性蛋白的抗体的免疫染色后使用光学显微镜检查来评估髓鞘形成的程度。这些方法部分揭示了PMD模型小鼠的髓鞘形成障碍。电镜检查显示髓鞘化程度适当。在中枢神经系统中，视神经因其神经元和少突胶质细胞数量均匀，在定量检测中具有上级优势。少

项目成果

Pelizaeus-Merzbacher disease

• DOI: 10.1093/jnen/61.9.747
• 发表时间: 2002-09-01
• 期刊: JOURNAL OF NEUROPATHOLOGY AND EXPERIMENTAL NEUROLOGY
• 影响因子: 3.2
• 作者: Koeppen, AH;Robitaille, Y
• 通讯作者: Robitaille, Y

Aberrant trafficking of a proteolipid protein in a mild Pelizaeus-Merzbacher disease

• DOI: 10.1016/j.neuroscience.2006.05.067
• 发表时间: 2006-01-01
• 期刊: NEUROSCIENCE
• 影响因子: 3.3
• 作者: Koizume, S.;Takizawa, S.;Osaka, H.
• 通讯作者: Osaka, H.

Aberrant trafficking of a mutated proteolipid protein in a mild Pelizaeus-Merzbacher disease.

轻度 Pelizaeus-Merzbacher 病中突变蛋白脂质蛋白的异常运输。

• 发表时间: 2006
• 作者: Koizume S;Takizawa S;Yamashita S;Miyagi Y;OsakaH
• 通讯作者: OsakaH

elizaeus-Merzbacher病; 小児中枢神経疾患の画像診断2008

Elizaeus-Merzbacher病；小儿中枢神经系统疾病的影像诊断2008年

• 发表时间: 2008
• 作者: Koizume S;Takizawa S;Yamashita S;Miyagi Y;OsakaH;小坂 仁
• 通讯作者: 小坂 仁

Vaccination and infection as causative factors in Japanese patients with Rasmussen syndrome: molecular mimicry and HLA class I.

• DOI: 10.1080/17402520600589522
• 发表时间: 2006-06
• 期刊: CLINICAL & DEVELOPMENTAL IMMUNOLOGY
• 作者: Takahashi, Yukitoshi;Matsuda, Kazumi;Kubota, Yuko;Shimomura, Jiro;Yamasaki, Etsuko;Kudo, Tatsuya;Fukushima, Katsuyuki;Osaka, Hitoshi;Akasaka, Noriyuki;Imamura, Atsushi;Yamada, Shinji;Kondo, Naomi;Fujiwara, Tateki
• 通讯作者: Fujiwara, Tateki