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.

哺乳动物耳蜗的治疗方法，包括耳蜗基因移植，已经用小鼠作为实验聋人模型进行了研究。随着小鼠基因组学的重大发展和具有良好特征的听力损失小鼠突变株的可用性，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入路进入kl/kl小鼠（klotho基因敲除小鼠）中耳和内耳的方法，而不是在其他啮齿动物中常规使用的耳后入路，并将其应用于耳蜗基因转移。这种腹侧入路可以通过腺病毒载体将klotho基因快速直接递送到klotho小鼠内耳，同时避免失血、面神经发病和死亡。3天后，在Corti器官、螺旋神经节细胞、螺旋韧带中检测到转基因表达，其模式与先前在豚鼠耳蜗中描述的相似。klotho基因成功进入并传递到kl/kl小鼠耳蜗，从而挽救了听力损失。本研究中发现的基因表达的复制将促进聋鼠在未来研究靶向耳蜗基因治疗中的应用。