Molecular Pathogenesis of Brain Damage and Gene Therapy in Genetic Leukodystrophy

1. 1)利用Krabbe globoid Cell Leukodystrophy （GLD）动物模型，试图找出抽搐小鼠白质营养不良的原因。斑痉挛法证实，神经细胞损伤的原因可能是细胞内钙的内流。细胞内钙的增加导致细胞蛋白酶的激活，从而对神经细胞造成损伤。2)我们研究了异色性脑白质营养不良的临床表型和基因型，并在2例MLD中发现了新的基因型。此外，我们还发现G99D突变为神经严重型，占日本mld所有基因型的50%。3)日本干燥拉尔松综合征患者具有日本特有的基因型。4)我们研究了法布里病的基因型鉴定，分别为L16H、A37V、W209X、342QIVS-1-1等。遗传白质营养不良的细胞治疗和基因治疗采用Twitcher小鼠和Sly小鼠进行基因治疗和细胞治疗。1)在胎儿期，将腺病毒载体注入Twitcher小鼠脑室内。注射病毒载体后，治疗动物的球状细胞数量减少。同时，治疗动物的精神素含量降低。2)将人胎脑提取的神经干细胞注射到Sly小鼠脑内，减少了Sly小鼠脑内积累的化合物。这些数据表明，神经干细胞对于治疗这些神经突变的中枢神经系统病变是有效的。3)间充质干细胞注入Sly小鼠后，其储存物质减少，对神经系统小鼠有效。

1. Pathogenesis of Leukodystrophy in Globoide Cell Leukodystrophy and Other1) Using animal model of Krabbe globoid cell leukodystrophy(GLD), we tried to identified the cause of leukodytrophy in twitcher mice. The cause of neural cell damage might be caused by the influx of intracellular calcium in such mice which was demonstrated by patch cramp method. The increased intracellular calcium resulted in the activation of cellular protease and hense damage the neural cells.2) We studied the clinical phnotype and genotype in metachromatic leukodystrophy and also identified novel genotype in two cases with MLD. Furthermore, we demonstrated that the G99D mutation was neruological severe type and consisted of 50% of all genotype of Japanese MLD.3) Japanese patients with Sjogren Larrson syndrome shows particular genotype in Japanese.4) We studied the genotype identifications in Fabry disease which were L16H, A37V, W209X, 342QIVS-1-1 etc.2. Cell therapy and gene therapy in genetic leukodystrophyGene therapy and cell therapy were carried out using Twitcher mice and Sly Mice.1) Twitcher mice were treated with adenovirus vector which was administered into intraventricle, during fetal period. The number of globoid cells were decreased in treated animals after the administration of viral vetor. Simultaneously, the amount of psychosine was decreased in treated animals.2) Neural stem cells obtained fromhuman fetal brains were injected into Sly mice brain and the accumulated compounds in Sly mice were decreased. The data suggest that neural stem cells were effective for the treatment of The CNS involvement in these neurological mutants.3) Injection of mesenchymal stem cells into Sly mice showed decreased storage Materials and effective for neurological mice.