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Rescue of hearing and vestibular function by antisense oligonucleotides in a mouse model of human deafness

Nature Medicine volume 19, pages 345–350 (2013)Cite this article

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Abstract

Hearing impairment is the most common sensory disorder, with congenital hearing impairment present in approximately 1 in 1,000 newborns 1 . Hereditary deafness is often mediated by the improper development or degeneration of cochlear hair cells 2 . Until now, it was not known whether such congenital failures could be mitigated by therapeutic intervention 3, 4, 5 . Here we show that hearing and vestibular function can be rescued in a mouse model of human hereditary deafness. An antisense oligonucleotide (ASO) was used to correct defective pre-mRNA splicing of transcripts from the USH1C gene with the c.216G>A mutation, which causes human Usher syndrome, the leading genetic cause of combined deafness and blindness 6, 7 . Treatment of neonatal mice with a single systemic dose of ASO partially corrects Ush1c c.216G>A splicing, increases protein expression, improves stereocilia organization in the cochlea, and rescues cochlear hair cells, vestibular function and low-frequency hearing in mice. These effects were sustained for several months, providing evidence that congenital deafness can be effectively overcome by treatment early in development to correct gene expression and demonstrating the therapeutic potential of ASOs in the treatment of deafness.

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Figure 1: Correction of USH1C 216A splicing using ASOs.
Figure 2: ASOs correct vestibular function and rescue hearing in 216AA mice.
Figure 3: ASO-29 treatment corrects mRNA splicing and harmonin protein expression and prevents cochlear hair cell loss in 216AA mice.
Figure 4: Restoration of hair cell stereocilia bundle shape in mice.

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Acknowledgements

We gratefully acknowledge support from the Hearing Health Foundation, Midwest Eye-Banks, the National Organization for Hearing Research Foundation, Capita Foundation and the US National Institutes of Health. We thank D. Cunningham and E. Rubel for assistance with scanning electron microscopy analysis; A. Rosenkranz, R. Marr and M. Oblinger for use of equipment; J. Huang for assistance with open-field analysis, L. Ochoa for assistance with ABR analysis computer support, G. MacDonald for assistance with confocal imaging and deconvolution analysis, U. Wolfrum (Johannes Gutenberg University of Mainz) for harmonin b–specific antibodies, H. Thompson for statistical analysis, and A. Case, B. Keats and M. Havens for discussions and comments on the manuscript.

Author information

Author notes

  1. Jennifer J Lentz, Francine M Jodelka and Anthony J Hinrich: These authors contributed equally to this work.

Authors and Affiliations

  1. Neuroscience Center and Department of Otorhinolaryngology & Biocommunications, Louisiana State University Health Sciences Center (LSUHSC), New Orleans, Louisiana, USA

    Jennifer J Lentz, Hamilton E Farris & Matthew J Spalitta

  2. Department of Cell Biology and Anatomy, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, Illinois, USA

    Francine M Jodelka, Anthony J Hinrich, Kate E McCaffrey & Michelle L Hastings

  3. Neuroscience Center and Department of Ophthalmology, LSUHSC, New Orleans, Louisiana, USA

    Nicolas G Bazan

  4. Department of Cellular and Molecular Pharmacology, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, Illinois, USA

    Dominik M Duelli

  5. Isis Pharmaceuticals, Carlsbad, California, USA

    Frank Rigo

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  1. Jennifer J Lentz
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Contributions

The project was conceived of by M.L.H. Experiments were designed and performed by A.J.H., F.M.J., J.J.L., K.E.M., M.L.H. and D.M.D., and were analyzed by M.L.H., J.J.L., F.R., F.M.J., A.J.H. and K.E.M. Animal work was carried out by M.L.H., D.M.D., K.E.M., A.J.H., F.M.J., J.J.L. and M.J.S. Molecular experiments were performed by A.J.H., F.M.J., K.E.M. and M.L.H. J.J.L. carried out the immunofluorescence analysis. J.J.L., M.J.S. and H.E.F. performed auditory brainstem response experiments, and J.J.L. and N.G.B. interpreted the results. M.L.H. and J.J.L. wrote the paper.

Corresponding authors

Correspondence to Jennifer J Lentz or Michelle L Hastings.

Ethics declarations

Competing interests

F.R. may materially benefit financially through stock options in Isis Pharmaceuticals. M.L.H. and F.R. have patents pending with the United States Patent and Trademark Office for the ASOs and the targeting approach.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–10 and Supplementary Table 1 (PDF 3338 kb)

Supplementary Video 1

Ush1c.216AA mice treated with ASO-29. This video shows four 1-month-old mice. Two representative heterozygous (Ush1c.216GA) mice (top row) and homozygous mutant mice (Ush1c.216AA) (bottom row), one of each group treated with ASO-C (left) and one with ASO-29 (right). The ASO-C treated mutant exhibits characteristic circling behavior and hyperactivity (bottom left), whereas the ASO-29-treated mouse (bottom right) is indistinguishable from the heterozygous mice (top). (MOV 2547 kb)

Supplementary Video 2

Behavioral response to acoustic stimuli in Ush1c.216AA mice treated with ASO-29. This video shows four 2-month-old mice. Two representative heterozygous (Ush1c.216GA) mice (top row) and homozygous mutant mice (Ush1c.216AA) (bottom row), one of each group treated with ASO-C (left) and one with ASO-29 (right). Opaque dividers were inserted between the cages to blind the mice to each other. A whistle blow (∼3-16 kHz and 90-110 dB SPL; Supplementary Fig. 6) was activated after acclimation to the environment. This acoustic stimulus occurs 2.9 s into the video and persists for 0.9 s. A startle response, indicated by a rapid body movement, followed by freezing (a period of watchful immobility), suggests, qualitatively, that the heterozygote mice (GA) and the mutant 216AA mouse treated with ASO-29 could hear the acoustic stimulation. The 216AA mutant mouse treated with ASO-C showed no visible response to the acoustic stimulus. (MOV 2776 kb)

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Lentz, J., Jodelka, F., Hinrich, A. et al. Rescue of hearing and vestibular function by antisense oligonucleotides in a mouse model of human deafness. Nat Med 19, 345–350 (2013). https://doi.org/10.1038/nm.3106

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