Gene delivery with adeno virus vector into the inner ear of deaf mouse

用腺病毒载体将基因导入聋小鼠内耳

• 批准号: 11671679
• 负责人: DOI Katsumi
• 金额: $ 2.5万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671679/
• 关键词: adeno virus vector; gene therapy; deafness; cochlea; アデノウイルスベクタ-; 内耳; 遺伝子導入; カリウムチャネル; トランスジェニック

Therapeutic manipulations of the mammalian cochlea, including cochlear gene transfer, have been studied using mouse as the experimental deaf model. With the significant developments in mouse genomics and the availability of mutant strains of mice with well-characterized hearing loss, the mouse will be preferred animal model for therapeutic manipulations. This study describes a ventral approach, instead of the routinely used postauricular approach in other rodents, for accessing the middle and inner ear of the kl/kl mouse (klotho gene knock-out mouse), and its application in cochlea gene transfer. This ventral approach enabled rapid and direct delivery of klotho gene with adeno virus vector to the klotho mouse inner ear while avoiding blood loss, facial nerve morbidity, and mortality. Transgene expression at 3 days was detected in the organ of Corti, spiral ganglion cells, spiral ligament, in a pattern similar to the previously described in the guinea pig cochlea. The successful access and delivery of klotho gene to the kl/kl mouse cochlea resulted in a rescue from hearing loss. The replication of gene expression seen in this study should promote the use of the deaf mouse in future studies investigating targeted cochlear gene therapy.

使用小鼠作为实验性聋模型研究了哺乳动物耳蜗的治疗操作，包括耳蜗基因转移。随着小鼠基因组学的重大发展以及具有良好特征的听力损失的小鼠突变株的出现，小鼠将成为治疗操作的首选动物模型。本研究描述了一种腹侧方法，而不是其他啮齿类动物常规使用的耳后方法，用于进入 kl/kl 小鼠（klotho 基因敲除小鼠）的中耳和内耳，及其在耳蜗基因转移中的应用。这种腹侧方法能够将带有腺病毒载体的 klotho 基因快速直接递送至 klotho 小鼠内耳，同时避免失血、面神经发病和死亡。 3天时在柯蒂氏器、螺旋神经节细胞、螺旋韧带中检测到转基因表达，其模式与之前在豚鼠耳蜗中描述的模式相似。 klotho 基因成功进入并传递至 kl/kl 小鼠耳蜗，从而挽救了听力损失。本研究中看到的基因表达的复制应该会促进聋鼠在未来研究靶向耳蜗基因治疗的研究中的使用。