Therapeutic effect of adeno-associated virus (AAV)-mediated ADNF-9 expression on cochlea of kanamycin-deafened guinea pigs

Abstract

Conclusion: rAAV-NT4-ADNF-9 could ameliorate the damage to auditory function and repair previous impairment of cochlear hair cell loss induced by kanamycin. Objective: To investigate the therapeutic effect of ADNF-9 on cochlear hair cells using the recombinant adeno-associated virus (AAV) carrying fusion gene NT4-ADNF-9 and the kanamycin-deafened guinea pig model. Methods: Forty white guinea pigs with normal auricle reflex and normal auditory brainstem responses (ABRs) were randomly divided into four groups. Kanamycin was administered to the animals in groups A, B, and C to establish the deafened guinea pig model. rAAV-NT4-ADNF-9, vector only, and artificial perilymph were then delivered to the cochlear tissue of animals in groups A, B, and C, respectively, through the round window membrane. Animals in group D did not receive any treatment and acted as normal controls. The hearing thresholds on the surgery side were recorded before and after the transfection treatment. Fourteen days after treatment, cochleae were removed for paraffin slide preparation and cochlear surface preparation. A phase contrast microscope was used to observe the protective effect of ADNF-9 on hair cells. Results: Significant reduction of the ABR threshold was observed after rAAV-NT4-ADNF-9 treatment (p < 0.05). After 14 days of treatment, the ABR threshold was also significantly different between the rAAV-NT4-ADNF-9-infected group and the non-infected group. Moreover, phase contrast microscopy showed significantly less hair cell damage or hair cell loss in the group treated with rAAV-NT4-ADNF-9 than in the groups treated with vector only or artificial perilymph (p < 0.05).

Keywords::

• Sensorineural deafness
• activity-dependent neurotrophic factor-9
• recombinant adeno-associated virus
• gene therapy

Acknowledgments

The study was supported by the National Natural Science Foundation of China (no. 81271068), the Research Fund for the Doctoral Program of Higher Education of China (20120201110060) and Scientific and Technological Project of Shaanxi Province (no. 2012K14-02-01), and the Youth Foundation of the Second Hospital of Xi`an jiaotong University (no. YJ(QN)200913). We thank supports from Tysta Skolan and Karolinska Institutet, Stockholm, Sweden

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.