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Studies of genetic analysis and gene therapy for myelin-associated disorders.

髓磷脂相关疾病的遗传分析和基因治疗研究。

基本信息

批准号：
04454282
负责人：
MAEKAWA Kihei
金额：
$ 3.71万
依托单位：
The Jikei University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

项目摘要

We investigated the genotype of Japanese Gaucher disease (GD), metachromatic leukodystrophy (MLD) to evaluate the correlation between genotype and phenotype, and to characterize the ethnicity. In 7 patients with type II GD, a new 754A missense mutation was identified in 3 cases heteroallelically.A new heteroalleic 1070A missense mutation was found in 2 cases of Japanese patients with MLD.These two mutations have not been reported mong Jewish and Caucasian population, which suggests that the genotypes of GD and MLD in Japan is unique.In order to evaluate the potential gene therapy for MLD, we constructed a retroviral vector for the transfer of the arylsulfatase cDNA, and transfected and expressed the gene in human MLD fibroblasts. As a result of gene transfer, the enzyme activity of MLD fibroblasts was increased about 70% of normal fibroblast level. Those observation demonstrates the feasibility of gene therapy for MLD.To understand the cause of demyelination in Krabbe's disease (GLD), the neurotoxicity of psychosine in neural cell culture was investigated. By immunofluorescence staining method using an anti-neurofilament antibody, psychosine treated cells showed destruction of cytoskelton and pathy intracellular changes. An electron microscopic study showed swelling of mitochondria, desruption of cristae in mitochondria, and disappearance of microtubules and neuro-filaments. These data suggest that psychosine influences mitochondrial function, possibly through inhibition in the destruction of cellular components.

我们调查了日本戈谢病（GD）和异染性脑白质营养不良（MLD）的基因型，以评估基因型和表型之间的相关性，并描述种族特征。在7例II型GD患者中，3例发现了新的754 A错义突变，2例日本MLD患者发现了新的1070 A错义突变，这两种突变在犹太人和高加索人群中未见报道，表明日本GD和MLD的基因型是独特的。我们构建了一个用于转移芳基硫酸酯酶cDNA的逆转录病毒载体，并在人MLD成纤维细胞中转染和表达该基因。由于基因转移的结果，MLD成纤维细胞的酶活性增加了约70%的正常成纤维细胞水平。为了解Krabbe病（GLD）脱髓鞘的原因，本研究在神经细胞培养中观察了精神病碱的神经毒性。用抗神经丝抗体免疫荧光染色法观察，精神病碱处理的细胞胞浆素被破坏，细胞内病变。电镜观察显示线粒体肿胀，嵴断裂，微管和神经丝消失。这些数据表明，精神病影响线粒体功能，可能是通过抑制细胞成分的破坏。