Multiple approaches of the therapy for lysosomal diseases affected with central nervous system

中枢神经系统溶酶体疾病的多种治疗方法

• 批准号: 13470164
• 负责人: SAKAI Norio
• 金额: $ 9.28万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470164/
• 关键词: lysosomal disease; psychosine; T cell death associated gene 8; Krabbe disease; Farber disease; saposin A; Molecular shaperon; ガラクトセレブロシダーゼ; 遺伝子変異解析; 蛋白質発現系; DHPLC法

Lysosomal diseases were caused by the accumulation of undigested substrates within lysosome. The lysosomal diseases affected in the central nervous system were hard to treat because the recombinant enzyme could not be entered through blood brain barrier. We tried several projects to approach the therapy of these disease, as follows ;1)Express affected enzyme: We tried to construct for the normal galactocerebrosidase to express in large-scale in yeast, however the ratio of expression was not good enough.2)Inhibition of synthetic enzyme of sphingolipid : We realize that cycloserine was confirmed to inhibit production of psychosine. We will check dose dependency for the effect of cycloserine.3)Molecular shaperon : Saposine A was proven to be the activator of galactocerebrosidase and might be shaperon for it. We were succeeded in express saposin A in yeast expression system. Purified saposin A was able to activate β-glucosidase and galactocerebrosidase.4)Activation of affected enzyme: The produced saposin A might be able to bind enzyme and stabilyze them. We are analyzing this effect in vivo.5)Block apoptosis cascade : We check the expression profile of TDAG8 and analyzed the effect of antibody against TDAG8.

溶酶体疾病是由溶酶体中无需困难的底物的积累引起的。在中枢神经系统中受影响的溶酶体疾病很难治疗，因为无法通过血液脑屏障输入重组酶。 We tried several projects to approach the therapy of these diseases, as follows ;1)Express affected enzyme: We tried to construct for the normal galactocerebrosidase to express in large-scale in yeast, however the ratio of expression was not good enough.2)Inhibition of synthetic enzyme of sphingolipid: We realized that cycloserine was confirmed to inhibit production of psychosine.We will check dose dependency for the effect 3）分子染色体：皂苷A已被证明是半乳脑酶酶的激活剂，可能是叶生成剂。我们在酵母表达系统中取得了成功的酵母表达剂表达saposin a。硫化皂苷a在酵母上表达。结合酶并将其稳定。我们在体内进行了分析。5）块凋亡级联反应：我们检查TDAG8的表达谱并分析了抗体对TDAG8的影响。

项目成果

Yanagihara I, Inui K, Yanagihara K, Park YD, Tanaka J, Ozono K, Okada S, Kurahashi H.: "Fluorescence in situ hybridization analysis of peripheral blood cells in Pearson marrow-pancreas syndrome."J Pediatr.. 139(3). 452-455 (2001)

Yanagihara I、Inui K、Yanagihara K、Park YD、Tanaka J、Ozono K、Okada S、Kurahashi H.：“皮尔逊骨髓-胰腺综合征外周血细胞的荧光原位杂交分析。”J Pediatr.. 139（3）

Kokubu C, Wilm C, Kokubu T, Walil M, Rodrigo I, Sakai N, Santagati F, Hayashizaki Y, Suzuki M, Yamamura K, Abe K, Imai K: "Undulated short-tail Deletion Mutation in the Mouse Ablates Paxi and Leads to Ectopic Activation of Neighboring NkxZ-2 in Domains th

Kokubu C、Wilm C、Kokubu T、Walil M、Rodrigo I、Sakai N、Santagati F、Hayashizaki Y、Suzuki M、Yamamura K、Abe K、Imai K：“小鼠中的波状短尾缺失突变消除了 Paxi 和引线

Tsukamoto H, Yamamoto T, Nishigaki T, Sakai N, Nanba E, Ninomiya H, Ohno K, Inui K, Okada S.: "T SSCP analysis by RT CR for the prenatal diagnosis of Niemann-Pick disease type C."Prenat Diagn. 21(1). 55-57 (2001)

Tsukamoto H、Yamamoto T、Nishigaki T、Sakai N、Nanba E、Ninomiya H、Ohno K、Inui K、Okada S.：“通过 RT CR 进行 T SSCP 分析，用于 C 型尼曼匹克病的产前诊断。”Prenat Diagn

Inui K, Akagi M, Ono J, Tsukamoto H, Shimono K, Mano T, Imai K, Yamada M, Muramatsu T, Sakai N, Okada S: "Mutational analysis of MECP2 in Japanese patients with atypical Rett syndrome"Brain Dev.. 23(4). 212-215 (2001)

Inui K、Akagi M、Ono J、Tsukamoto H、Shimono K、Mano T、Imai K、Yamada M、Muramatsu T、Sakai N、Okada S：“日本非典型 Rett 综合征患者 MECP2 的突变分析”Brain Dev..

Tsukamoto H, Yamamoto T, Nishigaki T, Sakai N, Inui K et al.: "SSCP analysis by RT-PCR for the prenatal diagnosis of Niemann-Pick disease type C"Prenat Diagn.. 21. 55-57 (2001)

Tsukamoto H、Yamamoto T、Nishigaki T、Sakai N、Inui K 等人：“通过 RT-PCR 进行 SSCP 分析用于 Niemann-Pick 病 C 型产前诊断”Prenat Diagn.. 21. 55-57 (2001)