Gene expression and its application to the investigation on pathogenesis of congenital metabolic diseases and development of therapy for them.

基因表达及其在先天性代谢性疾病发病机制研究和治疗开发中的应用。

• 批准号: 03670516
• 负责人: SAKURABA Hitoshi
• 金额: $ 1.34万
• 依托单位: The Tokyo Metropolitan Institute of Medical Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670516/
• 关键词: gene expression; Fabry disease; alpha-galactosidase; baculovirus; enzyme replacement therapy; confocal laser scanning microscopy; galactosialidosis; protective protein; ファブリー病; alpha-ガラクトシダーゼ; 先天代謝異常症; キメラ蛋白; レーザー走査型共焦点顕微鏡; ファブリ-病; αーガラクトシダ-

Effors were directed to clarify the pathogenesis of Fabry disease and galactosialidosis and develop the therapy for these diseases by means of gene expression system.Fabry disease is an X-linked inborn error of glycosphingolipid catabolism resulting from the deficient activity of alpha-galactosidase. The specific alpha-galactosidase mutations that cause the classic or variant Fabry disease phenotypes have been deterined. A variety of mutations, including deletions, nonsense mutations, splicing mutations and amino acid substitutions caused complete deficiency of alpha-galactosidase activity and resulted in the classic form of Fabry manifestations. Single base substitutions between 5'-end and the center of exon 6 led to the residual enzyme activity and variant form of Fabry phenotype. A large amount of human alpha-galactosidase was expressed using recombinant baculovirus/insect cell expression system and a possibility of enzyme replacement therapy for Fabry disease was investigated.A genetic defect of protective protein causes an systemic disease, galactosialidosis. Expression of human protective protein was established in transformed Chinese hamster ovary cells, and purified protective protein was confirmed to be a multifunctional enzyme protein.

Fabry病（Fabry disease）是由α-半乳糖苷酶（alpha-galactosidase）活性缺陷引起的X连锁遗传性鞘糖脂（glycosphingolipid，GSL）代谢异常。引起典型或变异型法布里病表型的特异性α-半乳糖苷酶突变已经确定。多种突变，包括缺失、无义突变、剪接突变和氨基酸取代，导致α-半乳糖苷酶活性完全缺乏，并导致典型形式的法布里病表现。第6外显子5 '端与中心之间的单碱基替换导致酶活性残留和Fabry表型变异。利用重组杆状病毒/昆虫细胞表达系统大量表达人α-半乳糖苷酶，并探讨其用于Fabry病的酶替代治疗的可能性。在转化的中国仓鼠卵巢细胞中表达了人保护蛋白，纯化的保护蛋白被证实是一种多功能酶蛋白。

项目成果

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Yoshida K.et al.：“GM1-神经节苷脂沉积症的表型-基因型相关性。”

Oshima A: "GM1 gangliosidosis:Tandem duplication within exon3 of β-galactosidase gene in an infantile patient." Clin.Genet.41. 235-238 (1992)

Oshima A：“GM1 神经节苷脂沉积症：婴儿患者 β-半乳糖苷酶基因外显子 3 内的串联重复。”Clin.Genet.41 (1992)。

Yoshida K: "Human β-galactosidase gene mutations in GM1-gangliosidosis:A common mutation among Japanese adult/chronic cases." Am.J.Hum.Genet.49. 435-442 (1991)

Yoshida K：“GM1-神经节苷脂沉积症中的人类 β-半乳糖苷酶基因突变：日本成人/慢性病例中的常见突变。Am.J.Hum.Genet.435-442 (1991)。

Nagao Y.: "Hypertrophic cardiomyopathy in late-onset variant of Fabry disease with high residual activity of α-galactosidaseA." Clin.Genet.39. 233-237 (1991)

Nagao Y.：“法布里病迟发型肥厚型心肌病，α-半乳糖苷酶 A 残留活性高。”Clin.Genet.39 (1991)。

Tsuji A: "Lysosomal enzyme replacement using α2-macroglobulin as a transport vehicle." J.Biochem.(in press). (1994)

Tsuji A：“使用 α2-巨球蛋白作为运输工具的溶酶体酶替代。”J.Biochem（出版中）。