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Topical application of the antiapoptotic TAT-FNK protein prevents aminoglycoside-induced ototoxicity

Abstract

We previously demonstrated that an artificial protein, TAT-FNK, has antiapoptotic effects against cochlear hair cell (HC) damage caused by ototoxic agents when applied systemically. To examine the feasibility of topical protein therapy for inner ear disorders, we investigated whether gelatin sponge soaked with TAT-FNK and placed on the guinea pig round window membrane (RWM) could deliver the protein to the cochlea and attenuate aminoglycoside (AG)-induced cochlear damage in vivo. First, we found that the immunoreactivity of TAT-myc-FNK was distributed throughout the cochlea. The immunoreactivity was observed from 1–24 h after application. When Tat-FNK was applied 1 h before ototoxic insult (a combination of kanamycin sulfate and ethacrynic acid), auditory brainstem response threshold shifts and the extent of HC death were significantly attenuated. When cochlear organotypic cultures prepared from P5 rats were treated with kanamycin, TAT-FNK significantly reduced the extent of caspase-9 activation and HC death. These findings indicate that TAT-FNK topically applied on the RWM can enter the cochlea by diffusion and effectively prevent AG-induced apoptosis of cochlear HCs by suppressing the mitochondrial caspase-9 pathway.

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Acknowledgements

We thank Ms A Tsuyuzaki and Ms Y Kurasawa (Department of Otolaryngology and Head and Neck Surgery, Faculty of Medicine, University of Tokyo, Tokyo, Japan) for technical assistance. This work was supported by Grants (17659527 and 20390440) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan to TY and a Grant (15110201) from the Ministry of Health, Labor and Welfare of Japan to TY.

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Kashio, A., Sakamoto, T., Kakigi, A. et al. Topical application of the antiapoptotic TAT-FNK protein prevents aminoglycoside-induced ototoxicity. Gene Ther 19, 1141–1149 (2012). https://doi.org/10.1038/gt.2011.204

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