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: 10557076
• 负责人: MATSUO Masafumi
• 金额: $ 8.7万
• 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: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10557076/
• 关键词: dystrophin; splicing; splicing enhancer sequence; exon skipping; treatment; ジストロフィン遺伝子; エクソン; スキッピング; アンチセンスオリゴオヌクレオチド

Duchenne muscular dystrophy (DMD) is a severe muscle wasting disease caused by mutation of the dystrophin gene. DMD patients usually die among the age of 20, but no treatment has been established. We have proposed that out-frame mutation identified in DMD can be corrected to in-frame by inducing exon skipping at the time of splicing. To confirm our proposal, we transfected oligonucleotide which is complementary to splicing enhancer sequence of exon 19 into cultured muscle cells, which was established from DMD case having exon 20 deletion. By this treatment, exon 19 skipping was induced and resulting dystrophin transcript re-gained translational reading frame. Remarkably dystrophin was stained in those transfected cells. This confirms that dystrophin negative cells are able to be converted to dystrophin positive and our proposal is right way to treat DMD.To expand our strategy, we are currently studying splicing enhancer sequence in other exons, dystrophin, splicing, splicing enhancer sequence, exon skipping, treatment

Duchenne型肌营养不良症（DMD）是由dystrophin基因突变引起的严重肌肉萎缩性疾病。DMD患者通常在20岁左右死亡，但尚未建立治疗方法。我们已经提出，DMD中鉴定的框外突变可以通过在剪接时诱导外显子跳跃来校正为框内突变。为了证实我们的提议，我们将与外显子19的剪接增强子序列互补的寡核苷酸转染到培养的肌细胞中，所述肌细胞是从具有外显子20缺失的DMD病例建立的。通过这种处理，诱导外显子19跳跃，并且所得肌营养不良蛋白转录物重新获得翻译阅读框架。在那些转染的细胞中显著地染色肌养蛋白。这证实了dystrophin阴性细胞能够转化为dystrophin阳性，我们的建议是治疗DMD的正确方法。为了扩展我们的策略，我们目前正在研究其他外显子中的剪接增强子序列，dystrophin，剪接，剪接增强子序列，外显子跳跃，治疗

项目成果

Dwi Pramono,ZA, Takeshima,Y, Surono,A, Ishida,T and Matsuo,M.: "A novel cryptic exon in intron 2 of the human dystrophin gene evolved from an intron by acquiring consensus sequence for splicing at different stages of anthropoid evolution"Biochem Biophys R

Dwi Pramono,ZA、Takeshima,Y、Surono,A、Ishida,T 和 Matsuo,M.：“人类肌营养不良蛋白基因内含子 2 中的一个新的神秘外显子是通过在类人猿的不同阶段获得剪接的共有序列而从内含子进化而来的

Wibawa,T, Takeshima,Y, Mitsuyoshi,H., Surono,A, Nakamura,H and Matsuo,M.: "Complete skipping of exon 66 due to novel mutation of the dystrophin gene was identified in two Japanese families of DMD with severe mental retardation"Brain Dev. (in press). (2000

Wibawa,T、Takeshima,Y、Mitsuyoshi,H.、Surono,A、Nakamura,H 和 Matsuo,M.：“由于抗肌营养不良蛋白基因的新突变，在两个患有严重 DMD 的日本家族中发现了外显子 66 的完全跳跃。

Chen,D,Takeshima,Y,Ishikawa,Y,Ishikawa,Y,Minami,R,Matsuo,M.: "A novel deletion of the dystrophin S-promoter region co-segregating with mental retardation." Neurology. (in press). (1999)

Chen,D,Takeshima,Y,Ishikawa,Y,Ishikawa,Y,Minami,R,Matsuo,M.：“抗肌营养不良蛋白 S 启动子区域的新型缺失与智力障碍共分离。”

Shiga,N,Matsuo,M,Yokoyama,M,Yokota,Y.: "Study on mutations affecting the muscle promoter/first exon of the dystrophin gene in 92 Japanese dilated cardiomyopathy patients." Am J Med Genet.79. 226-227 (1998)

Shiga,N,Matsuo,M,Yokoyama,M,Yokota,Y.：“影响 92 名日本扩张型心肌病患者肌营养不良蛋白基因的肌肉启动子/第一个外显子的突变研究。”

Shiga, N., Matsuo, M., Yokoyama, M and Yokota, Y.: "Study on mutations affecting the muscle promoter/first exon of the dystrophin gene in 92 Japanese dilated cardiomyopathy patients"Am. J. Med. Genet.. 79. 226-227 (1998)

Shiga, N.、Matsuo, M.、Yokoyama, M 和 Yokota, Y.：“影响 92 名日本扩张型心肌病患者肌营养不良蛋白基因肌肉启动子/第一外显子的突变研究”Am。