Establishment of treatment of Duchenne muscular dystrophy

杜氏肌营养不良症治疗方法的建立

• 批准号: 12557068
• 负责人: MATSUO Masafumi
• 金额: $ 8.58万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557068/
• 关键词: Duchenne muscular dystrophy; splicing; splicing enhancer sequence; exon skipping; antisense oligonucleotide; スキッピング; アンチセンス; スプライシング促進配列; スプライシング

It has been proposed that Duchenne muscular dystrophy can be treated with modification of out-of-frame mutation into in-frame mutation by inducing exon skipping. In this study the possibility to treat DMD was evaluated by seeing inducibility of exon skipping. In the previous study we showed exon 19 skipping can be induced by blocking the function of splicing enhancer sequence with antisense oligonucleotides. To apply this strategy we looked for splicing enhancer sequence in exons that are located in deletion hot spots. From exon sequence purine-rich sequence were extracted as candidate sequences for splicing enhancer sequence and splicing enhancer activity of those candidate sequences were analyzed by using Drosophilla dsx minigene. Thereby some antisense oligonucleotides were found to have an ability to induce exon skipping.

已经提出，Duchenne肌营养不良症可以通过诱导外显子跳跃将框外突变修饰为框内突变来治疗。本研究通过观察外显子跳跃的诱导作用来评估治疗DMD的可能性。我们在前期的研究中发现，用反义寡核苷酸阻断剪接增强子序列的功能可以诱导外显子19跳跃。为了应用这种策略，我们在位于缺失热点的外显子中寻找剪接增强子序列。从外显子序列中提取富含嘌呤的序列作为剪接增强子序列的候选序列，并利用果蝇dsx minigene分析这些候选序列的剪接增强子活性。因此，发现一些反义寡核苷酸具有诱导外显子跳跃的能力。

项目成果

Ito, T, Takeshima, Y, Nakamura, H, Matsuo, M.: "One of three examined purine-rich sequences selected from dystrophin exons exhibits splicing enhancer activity"Acta Myologica. 20. 151-153 (2001)

Ito, T, Takeshima, Y, Nakamura, H, Matsuo, M.：“从抗肌营养不良蛋白外显子中选出的三个经过检查的富含嘌呤的序列之一表现出剪接增强子活性”Acta Myologica。

Ito T.: "One of three examined purine-rich sequence selected from dystrophin exons exhibits splicing enhancer activity"Acta Myologica. 2. 151-153 (2001)

Ito T.：“从肌营养不良蛋白外显子中选择的三个经检查的富含嘌呤的序列之一表现出剪接增强子活性”Acta Myologica。

Matsuo M.: "Treatment of Duchenne muscular dystrophy at the mRNA level"Frontiers in human genetics diseases and technologies. 347-361 (2001)

Matsuo M.：“在 mRNA 水平上治疗杜氏肌营养不良症”人类遗传疾病和技术的前沿。

Takeshima Y.: "Nakamura, H, Matsuo, M. Oligonucleotides against a splicing enhancer sequence led to dystrophin production in muscle cells from a Duchenne muscular dystrophy patient."Brain Dev.. 23. 788-798 (2001)

Takeshima Y.：“Nakamura, H, Matsuo, M.针对剪接增强子序列的寡核苷酸导致杜氏肌营养不良症患者的肌肉细胞中产生肌营养不良蛋白。”Brain Dev.. 23. 788-798 (2001)

Suminaga,R.,: "Nonsense mutation of the alpha-actinin-3 gene is not associated with dystrophinopathy."Am.J.Med.Genet.. 92(1). 77-78 (2000)

Suminaga, R.，：“α-actinin-3 基因的无义突变与肌营养不良症无关。”Am.J.Med.Genet.. 92(1)。