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An in vitro model system to study gene therapy in the human inner ear

Gene Therapy volume 14, pages 1121–1131 (2007)Cite this article

Abstract

The confined fluid-filled labyrinth of the human inner ear presents an opportunity for introduction of gene therapy reagents designed to treat hearing and balance dysfunction. Here we present a novel model system derived from the sensory epithelia of human vestibular organs and show that the tissue can survive up to 5 days in vitro. We generated organotypic cultures from 26 human sensory epithelia excised at the time of labyrinthectomy for intractable Meniere's disease or vestibular schwannoma. We applied multiply deleted adenoviral vectors at titers between 105 and 108 viral particles/ml directly to the cultures for 4–24 h and examined the tissue 12–96 h post-transfection. We noted robust expression of the exogenous transgene, green fluorescent protein (GFP), in hair cells and supporting cells suggesting both were targets of adenoviral transfection. We also transfected cultures with a vector that carried the genes for GFP and KCNQ4, a potassium channel subunit that causes dominant-progressive hearing loss when mutated. We noted a positive correlation between GFP fluorescence and KCNQ4 immunolocalization. We conclude that our in vitro model system presents a novel and effective experimental paradigm for evaluation of gene therapy reagents designed to restore cellular function in patients who suffer from inner ear disorders.

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Acknowledgements

We thank Dr Thomas Jentsch and Dr Kirk Biesel for providing the coding sequence of the human and mouse KCNQ4 genes, respectively. We also thank Dr Andrea Amalfitano for providing Ad2-GFP, Dr Bechara Kachar for the KCNQ4 antibody and Dr Tama Hasson for the myosin-VIIa antibody. This work was supported by the NIH/NIDCD Grant #DC005439 to JR Holt, the Virginia Lion's Hearing Foundation and a Triological Society Career Development Grant to BW Kesser.

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Authors and Affiliations

  1. Department of Otolaryngology – Head and Neck Surgery, University of Virginia School of Medicine, Charlottesville, VA, USA

    B W Kesser, G T Hashisaki, K Fletcher & J R Holt

  2. Department of Neuroscience, University of Virginia School of Medicine, Charlottesville, VA, USA

    H Eppard & J R Holt

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  1. B W Kesser
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Correspondence to J R Holt.

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Supplementary Information accompanies the paper on Gene Therapy Web site (http://www.nature.com/gt)

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Kesser, B., Hashisaki, G., Fletcher, K. et al. An in vitro model system to study gene therapy in the human inner ear. Gene Ther 14, 1121–1131 (2007). https://doi.org/10.1038/sj.gt.3302980

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