development of gene therapy for auditory and vestibular disorders-gene delivery into the cochlear

听觉和前庭疾病基因治疗的发展-基因递送到耳蜗

• 批准号: 12557142
• 负责人: DOI Katsumi
• 金额: $ 7.81万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557142/
• 关键词: adeno virus vector; HVJ-liposome; sensorineural hearing loss; gene delivery; klotho gene; 血管条; コルチ器; ラセン神経節細胞; 呼吸上皮; 水チャネル; 遺伝子導入; 鼻粘膜

Auditory function of klotho gene knock out (kl/kl) mouse was evaluated by measuring of ABR and EP of kl/kl mouse was significantly deteriorated compared with those of wild mouse, suggesting that kl/kl mouse has sensorineural hearing loss. RT-PCR and immunohistochemical analysis indicated that there was no expression of klotho mRNA and protein in the cochlea. The results suggest that klotho protein secreted from other tissues must reach to the cochlea via blood circulation and exert its physiological action there as a humoral factor essential for normal auditory function. Intra-cochlear and intra-cutaneous gene delivery of normal klotho gene with adeno virus vector (AVV) was found to be able to rescue completely the auditory function of kl/kl mouse. Gene delivery into the cochlea with HVJ-lipose was also found to be effective.

与野生小鼠相比，通过测量KL/KL小鼠的ABR和EP的测量来评估Klotho Gene敲出（KL/KL）小鼠的听觉功能，这表明KL/KL小鼠具有感觉到听力损失。 RT-PCR和免疫组织化学分析表明，耳蜗中没有klotho mRNA和蛋白质表达。结果表明，从其他组织分泌的klotho蛋白必须通过血液循环到达耳蜗，并将其生理作用作为正常听觉功能必不可少的体液因子发挥作用。发现具有adeno病毒载体（AVV）正常klotho基因的近形和二骨内基因递送能够完全挽救KL/KL小鼠的听觉功能。还发现，用HVJ脂肪递送基因将基因递送到耳蜗有效。