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

以小鼠为耳聋实验模型，研究了包括耳蜗基因转移在内的哺乳动物耳蜗病的治疗方法。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。本研究介绍了KL/KL小鼠(Klotho基因敲除小鼠)的中耳和内耳的腹侧入路，以替代其他啮齿类动物常规使用的耳后入路，以及它在耳蜗基因转移中的应用。这种腹侧方法使Klotho基因和腺病毒载体能够快速和直接地传递到Klotho小鼠内耳，同时避免失血、面神经疾病和死亡。3天后，转基因在Corti器官、螺旋神经节细胞、螺旋韧带中均有表达，其表达模式与之前在豚鼠耳蜗组织中的表达模式相似。Klotho基因成功导入KL/KL小鼠耳蜗内，使其免于听力损失。这项研究中所看到的基因表达的复制应该会促进聋人小鼠在未来研究耳蜗靶向基因治疗的研究中的使用。

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.