Otof-/-成年鼠基因治疗DPOAE和ABR阈值变化研究
Study on gene therapy for DPOAE and ABR threshold changes in adult Otof-/-mice
王子菁 1曹麒 1胡少伟 2范新泰 2吕俊 2王会 2王武庆 2李华伟 2舒易来1
作者信息
- 1. 南华大学附属第二医院耳鼻喉科(湖南衡阳,421001);复旦大学附属眼耳鼻喉科医院耳鼻喉科
- 2. 复旦大学附属眼耳鼻喉科医院耳鼻喉科
- 折叠
摘要
目的:通过分析Otof-/-成年鼠在基因治疗前后各频率畸变产物耳声发射(distortion product otoa-coustic emissions,DPOAE)和听觉脑干反应(auditory brainstem response,ABR)的阈值变化及其相关性,探索治疗后听力恢复的评价方法.方法:经圆窗膜(round window membrane,RWM)路径,向4周龄Otof-/-成年鼠内耳注射双AAV载体携带的内含肽介导的OTOF基因治疗体系,检测治疗前后DPOAE与ABR阈值,免疫荧光染色评估耳畸蛋白(otoferlin)恢复表达的内耳毛细胞比例及突触数量,并进行统计学分析.结果:AAV-PHP.eB对耳蜗内毛细胞具有高转染率;治疗体系纠正了 Otof-//-成年鼠的听力,且不影响野生型小鼠听功能.基因治疗后DPOAE阈值与ABR阈值变化在16 kHz处存在显著相关;术后DPOAE略微上升,但在术后2个月时出现恢复趋势.结论:基因治疗可显著恢复Otof-/-成年鼠听力,基因治疗体系手术给药可能会引起听力损伤,这需要更加精细、轻柔的操作,以最大程度发挥基因治疗的作用.
Abstract
Objective:This study aims to analyze the threshold changes in distortion product otoacoustic emis-sions(DPOAE)and auditory brainstem response(ABR)in adult Otof-/-mice before and after gene therapy,e-valuating its effectiveness and exploring methods for assessing hearing recovery post-treatment.Methods:At the age of 4 weeks,adult Otof-/-mice received an inner ear injection of a therapeutic agent containing intein-media-ted recombination of the OTOF gene,delivered via dual AAV vectors through the round window membrane(RWM).Immunofluorescence staining assessed the proportion of inner ear hair cells with restored otoferlin ex-pression and the number of synapses.Statistical analysis was performed to compare the DPOAE and ABR thresh-olds before and after the treatment.Results:AAV-PHP.eB demonstrates high transduction efficiency in inner ear hair cells.The therapeutic regimen corrected hearing loss in adult Otof-/-mice without impacting auditory func-tion in wild-type mice.The changes in DPOAE and ABR thresholds after gene therapy are significantly correlated at 16 kHz.Post-treatment,a slight increase in DPOAE was observeds,followed by a recovery trend at 2 months post-treatment.Conclusion:Gene therapy significantly restored hearing in adult Otof-/-mice,though the surgi-cal delivery may cause transient hearing damage.Precise and gentle surgical techniques are essential to maximize gene therapy's efficacy.
关键词
OTOF/基因治疗/畸变产物耳声发射/听觉脑干反应Key words
OTOF/gene therapy/distortion product otoacoustic emissions/auditory brainstem response引用本文复制引用
基金项目
国家自然科学基金()
国家杰出青年科学基金(82225014)
国家重点研发计划(2020YFA0908201)
湖南省科技创新计划(2023RC4005)
出版年
2024
NETL
NSTL
万方数据