Study on the treatment of Duchenne musclar dystrophy with chimera RNA/DNA

嵌合体RNA/DNA治疗杜氏肌营养不良症的研究

• 批准号: 13307026
• 负责人: MATSUO Masafumi
• 金额: $ 35.28万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 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/grant/KAKENHI-PROJECT-13307026/
• 关键词: dystrophin; Duchenne; muscular dystrophy; molecular therapy; 遺伝子異常; デュシェンヌ型筋ジストロフィー; キメラRNA; DNA; 点変異; 筋細胞; 治療; 点突然変異

In this study the method to treat Duchenne musclar dystrophy (DMD) was examined by inducing exon 41 skipping. In one DMD a nonsense mutation was identified in exon 41 of the dystrophin gene. It was supposed that he will start dystrophin production when exon 41 skipping was induced resulting in in-frame mRNA. RNA/ENA chimera was employed to induce exon 41 skipping. Several RNA/ENA chimera were designed to cover the full-length of exon 41 and each of RNA/ENA chimera examined for its activity to induce exon 41 skipping. As a result the best RNA/ENA was identified. The selected RNA/ENA chimera was transfected to myocytes of the DMD case. Remarkably, exon 41 skipping was induced in his dystrophin mRNA and dystrophin expression was shown.

本研究探讨了诱导第41外显子跳脱治疗杜氏肌营养不良症（DMD）的方法。在一个DMD中，在肌营养不良蛋白基因的41号外显子上发现了无义突变。据推测，当41号外显子跳跃性被诱导产生帧内mRNA时，他将开始产生肌营养不良蛋白。采用RNA/ENA嵌合体诱导41外显子跳变。设计了多个RNA/ENA嵌合体覆盖41号外显子的全长，并检测了每个RNA/ENA嵌合体诱导41号外显子跳跃的活性。最终鉴定出最佳RNA/ENA。将选择的RNA/ENA嵌合体转染到DMD病例的肌细胞中。值得注意的是，肌营养不良蛋白mRNA外显子41被诱导跳跃，并显示出肌营养不良蛋白的表达。

项目成果

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

Matsuo M.：“在 mRDA 水平上治疗杜氏肌营养不良症。”人类遗传疾病和技术前沿.. 347-361 (2001)

Ito T.: "One of three examined purine-rich sequences 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检查的富含嘌呤序列之一表现出剪接增强子活性”Acta Myologica。

Adachi K, Takeshima Y, Wada H, Yagi Y, Nakamura H, Matsuo M.: "Hetergpus dystrophin mRNAs produced by a novel splice acceptor site mutation in intermediate dystrophinopathy."Ped.Res.. 53. 1-7 (2003)

Adachi K、Takeshima Y、Wada H、Yagi Y、Nakamura H、Matsuo M.：“中间型肌营养不良症中新型剪接受体位点突变产生的 Hetergpus 肌营养不良蛋白 mRNA。”Ped.Res.. 53. 1-7 (2003)

Nakayama Y: "Cloning of cDNA Encoding a Regeneration-associated Muscle Protease Whose Expression is Attenuated in Cell Lines Derived from Duchenne Muscular Dystrophy Patients"American Journal of Pathology. in press. (2004)

Nakayama Y：“克隆编码再生相关肌肉蛋白酶的 cDNA，该蛋白酶的表达在杜氏肌营养不良症患者的细胞系中减弱”，《美国病理学杂志》。

Ito T.: "Purine-rich exon sequences are not necessarily splicing enhancer sequence in the dystrophin gene."Kobe. J. Med. Sci.. 47. 193-202 (2001)

Ito T.：“富含嘌呤的外显子序列不一定是肌营养不良蛋白基因中的剪接增强子序列。”Kobe。