Letter | Published:

Treatment of autosomal dominant hearing loss by in vivo delivery of genome editing agents

Nature volume 553, pages 217221 (11 January 2018) | Download Citation

Abstract

Although genetic factors contribute to almost half of all cases of deafness, treatment options for genetic deafness are limited1,2,3,4,5. We developed a genome-editing approach to target a dominantly inherited form of genetic deafness. Here we show that cationic lipid-mediated in vivo delivery of Cas9–guide RNA complexes can ameliorate hearing loss in a mouse model of human genetic deafness. We designed and validated, both in vitro and in primary fibroblasts, genome editing agents that preferentially disrupt the dominant deafness-associated allele in the Tmc1 (transmembrane channel-like gene family 1) Beethoven (Bth) mouse model, even though the mutant Tmc1Bth allele differs from the wild-type allele at only a single base pair. Injection of Cas9–guide RNA–lipid complexes targeting the Tmc1Bth allele into the cochlea of neonatal Tmc1Bth/+ mice substantially reduced progressive hearing loss. We observed higher hair cell survival rates and lower auditory brainstem response thresholds in injected ears than in uninjected ears or ears injected with control complexes that targeted an unrelated gene. Enhanced acoustic startle responses were observed among injected compared to uninjected Tmc1Bth/+ mice. These findings suggest that protein–RNA complex delivery of target gene-disrupting agents in vivo is a potential strategy for the treatment of some types of autosomal-dominant hearing loss.

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Acknowledgements

This work was supported by DARPA HR0011-17-2-0049 (to D.R.L.), US NIH R01 EB022376 (to D.R.L.), R35 GM118062 (to D.R.L.), R01 DC006908 (to Z.-Y.C), P30 DC05209 (to M.C.L.), R01 DC00138 (to M.C.L.), and R01 DC013521 (to J.R.H.). We are grateful for support from the David-Shulsky Foundation (to Z.-Y.C.), a Frederick and Ines Yeatts Hair Cell Regeneration grant (to Y.T., V.L., M.H., and Y.S.), the Bertarelli Foundation and the Jeff and Kimberly Barber Fund (to J.R.H.), the Broad Institute (to D.R.L and Z.-Y.C.), and the HHMI (to D.R.L.). We thank H. Rees, S. Tsai, M. Packer, K. Zhao and D. Usanov for assistance.

Author information

Author notes

    • Xue Gao
    •  & Yong Tao

    Present addresses: Department of Chemical and Biomolecular Engineering, Rice University, Houston, Texas 77005, USA (X.G.); Department of Otolaryngology-Head and Neck Surgery, Shanghai Ninth People’s Hospital and Ear Institute, Shanghai Jiaotong University School of Medicine, Shanghai, 200011, China (Y.T.).

    • Xue Gao
    •  & Yong Tao

    These authors contributed equally to this work.

    • Zheng-Yi Chen
    •  & David R. Liu

    These authors jointly supervised this work.

Affiliations

  1. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA

    • Xue Gao
    • , Wei-Hsi Yeh
    • , Johnny H. Hu
    • , David B. Thompson
    •  & David R. Liu
  2. Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts 02138, USA

    • Xue Gao
    • , Wei-Hsi Yeh
    • , Johnny H. Hu
    • , David B. Thompson
    •  & David R. Liu
  3. Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02141, USA

    • Xue Gao
    • , Wei-Hsi Yeh
    • , Johnny H. Hu
    •  & David R. Liu
  4. Department of Otolaryngology and Program in Neuroscience, Harvard Medical School and Eaton Peabody Laboratory, Massachusetts Eye and Ear Infirmary, Boston, Massachusetts 02114, USA

    • Yong Tao
    • , Veronica Lamas
    • , Mingqian Huang
    • , Yu-Juan Hu
    • , Yilai Shu
    • , Hongyang Wang
    • , Daniel B. Polley
    • , M. Charles Liberman
    •  & Zheng-Yi Chen
  5. Department of Otorhinolaryngology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, China

    • Yong Tao
    • , Yu-Juan Hu
    •  & Wei-Jia Kong
  6. Program in Speech and Hearing Bioscience and Technology, Harvard University, Cambridge, Massachusetts 02138, USA

    • Wei-Hsi Yeh
  7. Departments of Otolaryngology and Neurology, F.M. Kirby Neurobiology Center Boston Children’s Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA

    • Bifeng Pan
    •  & Jeffrey R. Holt
  8. Department of Otolaryngology–Head and Neck Surgery, Eye and ENT Hospital, Shanghai Medical College, Fudan University, Shanghai, China

    • Yilai Shu
  9. Department of Biomedical Engineering, Tufts University, Medford, Massachusetts 02155, USA

    • Yamin Li
    •  & Qiaobing Xu
  10. Department of Otolaryngology & Head Neck Surgery, Key Lab of Hearing Impairment Science of Ministry of Education, Key Lab of Hearing Impairment Prevention and Treatment of Beijing City, Chinese PLA Medical School, Beijing, China

    • Hongyang Wang
    •  & Shiming Yang

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Contributions

X.G. and Y.T. designed the research, performed genome editing (X.G.) and hearing biology (Y.T.) experiments, analysed data, and wrote the manuscript. V.L., M.H., W.-H.Y., B.P., Y.-J.H., and H.W. designed experiments, performed hearing biology experiments and analysed data. D.B.P., M.C.L., and W.-J.K. designed hearing experiments and analysed data. Y.S. and S.Y. supported hearing biology experiments. J.H.H. analysed GUIDE-seq data. D.B.T. supported genome editing experiments. Y.L. and Q.X. designed and synthesized lipids. J.R.H., Z.-Y.C. and D.R.L. designed and supervised the research, and wrote the manuscript. All authors edited the manuscript.

Competing interests

D.R.L. is a consultant and co-founder of Editas Medicine, Beam Therapeutics, and Pairwise Plants, companies that use genome editing. X.G., D.B.T., Z.-Y.C., and D.R.L. have filed patent applications on aspects of this work.

Corresponding authors

Correspondence to Zheng-Yi Chen or David R. Liu.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

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    Life Sciences Reporting Summary

  2. 2.

    Supplementary Information

    This file contains a representative example of the gating strategy used in the flow cytometry experiments in Extended Data Figure 1a.

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    Supplementary Information

    This file contains supplementary methods which includes tables 1 and 2, sequences, notes and figure 1.

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https://doi.org/10.1038/nature25164

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