Skip to main content
SpringerLink
Log in

AAV-Mediated Gene Delivery to the Inner Ear

  • Protocol
  • First Online:
Adeno-Associated Virus Vectors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1950))

Abstract

Cochlear gene therapy has made tremendous strides over the past 5 years. The first study documenting successful restoration of congenital hearing loss using AAV-mediated gene therapy occurred in a mouse model of deafness lacking vesicular glutamate transporter 3 (VGLUT 3). This study utilized a trans-bulla round window membrane (RWM) delivery approach. Since this study, these methodologies have been applied to a number of other mouse models of genetic deafness with varying degrees of success, lending promise for future clinical application of this burgeoning technology. Here we describe a method of virally mediated gene delivery into the cochlear scala tympani through the RWM. This method involves negligible damage to essential structures of the middle and inner ear while preserving hearing. The efficacy of this surgical technique will be demonstrated by the restoration of hearing to the VGLUT3 knockout mice (a mouse model of congenital deafness) after delivery of VGLUT3 gene to the inner ear using an adeno-associated virus as a vector.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Kral A, O’Donoghue GM (2010) Profound deafness in childhood. N Engl J Med 363(15):1438–1450

    Article  CAS  PubMed  Google Scholar 

  2. Di Domenico M, Ricciardi C, Martone T, Mazzarella N, Cassandro C, Chiarella G, D’Angelo L, Cassandro E (2011) Towards gene therapy for deafness. J Cell Physiol 226(10):2494–2499

    Article  PubMed  Google Scholar 

  3. Shearer AE, Hildebrand MS, Sloan CM, Smith RJ (2011) Deafness in the genomics era. Hear Res 282(1–2):1–9

    Article  PubMed  PubMed Central  Google Scholar 

  4. Petersen MB, Willems PJ (2006) Non-syndromic, autosomal-recessive deafness. Clin Genet 69(5):371–392

    Article  CAS  PubMed  Google Scholar 

  5. Izumikawa M, Minoda R, Kawamoto K, Abrashkin KA, Swiderski DL, Dolan DF, Brough DE, Raphael Y (2005) Auditory hair cell replacement and hearing improvement by Atoh1 gene therapy in deaf mammals. Nat Med 11:271–276

    Article  CAS  PubMed  Google Scholar 

  6. Praetorius M, Hsu C, Baker K, Brough DE, Plinkert P, Staecker H (2009) Adenovector-mediated hair cell regeneration is affected by promoter type. Acta Otolaryngol 130(2):215–222

    Article  Google Scholar 

  7. Akil O, Seal RP, Burke K, Wang C, Alemi A, During M, Edwards RH, Lustig LR (2012) Restoration of hearing in the VGLUT3 knockout mouse using virally mediated gene therapy. Neuron 75(2):283–293

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Lentz JJ, Jodelka FM, Hinrich AJ, McCaffrey KE, Farris HE, Spalitta MJ, Bazan NG, Duelli DM, Rigo F, Hastings ML (2013) Rescue of hearing and vestibular function by antisense oligonucleotides in a mouse model of human deafness. Nat Med 19(3):345–350

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Staecker H, Schlecker C, Kraft S, Praetorius M, Hsu C, Brough DE (2014) Optimizing atoh1-induced vestibular hair cell regeneration. Laryngoscope 124:S1–S12

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Ryan AF, Mullen LM, Doherty JK (2009) Cellular targeting for cochlear gene therapy. Adv Otorhinolaryngol 66:99–115

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Xia L, Yin S, Wang J (2012) Inner ear gene transfection in neonatal mice using adeno-associated viral vector: a comparison of two approaches. PLoS One 7(8):e43218

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Bennett J, Ashtari M, Wellman J, Marshall KA, Cyckowski LL, Chung DC, McCague S, Pierce EA, Chen Y, Bennicelli JL, Zhu X, Ying GS, Sun J, Wright JF, Auricchio A, Simonelli F, Shindler KS, Mingozzi F, High KA, Maguire AM (2012) AAV2 gene therapy readministration in three adults with congenital blindness. Sci Transl Med 4(120):120ra15

    Article  PubMed  PubMed Central  Google Scholar 

  13. Simonelli F, Maguire AM, Testa F, Pierce EA, Mingozzi F, Bennicelli JL, Rossi S, Marshall K, Banfi S, Surace EM, Sun J, Redmond TM, Zhu X, Shindler KS, Ying GS, Ziviello C, Acerra C, Wright JF, McDonnell JW, High KA, Bennett J, Auricchio A (2010) Gene therapy for Leber’s congenital amaurosis is safe and effective through 1.5 years after vector administration. Mol Ther 18(3):643–650

    Article  CAS  PubMed  Google Scholar 

  14. Husseman J, Raphael Y (2009) Gene therapy in the inner ear using adenovirus vectors. Adv Otorhinolaryngol 66:37–51

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Ballana E, Wang J, Venail F, Estivill X, Puel JL, Arbonès ML, Bosch A (2008) Efficient and specific transduction of cochlear supporting cells by adeno-associated virus serotype 5. Neurosci Lett 442(2):134–139

    Article  CAS  PubMed  Google Scholar 

  16. Praetorius M, Brough DE, Hsu C, Plinkert PK, Pfannenstiel SC, Staecker H (2009) Adenoviral vectors for improved gene delivery to the inner ear. Hear Res 248(1–2):31–38

    Article  CAS  PubMed  Google Scholar 

  17. Kay MA, Glorioso CG, Naldini L (2001) Viral vectors for gene therapy: the art of turning infectious agents into vehicles of therapeutics. Nat Med 7(1):33–40

    Article  CAS  PubMed  Google Scholar 

  18. Seal RP, Akil O, Yi E, Weber CM, Grant L, Yoo J, Clause A, Kandler K, Noebels JL, Glowatzki E, Lustig LR, Edwards RH (2008) Sensorineural deafness and seizures in mice lacking vesicular glutamate transporter 3. Neuron 57:263–275

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Askew C, Rochat C, Pan B, Asai Y, Ahmed H, Child E, Schneider BL, Aebischer P, Holt JR (2015) Tmc gene therapy restores auditory function in deaf mice. Sci Transl Med 7(295):295ra108

    Article  PubMed  Google Scholar 

  20. Emptoz A, Michel V, Lelli A, Akil O, Boutet de Monvel J, Lahlou G, Meyer A, Dupont T, Nouaille S, Ey E, Franca de Barros F, Beraneck M, Dulon D, Hardelin JP, Lustig L, Avan P, Petit C, Safieddine S (2017) Local gene therapy durably restores vestibular function in a mouse model of Usher syndrome type 1G. Proc Natl Acad Sci U S A 114(36):9695–9700

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Geng R, Akil O, Gopal SR, Chen DH, Stepanyan R, Basch ML, Dinculescu A, Furness DN, Saperstein D, Hauswirth W, Lustig LR, Alagramam KN (2017) Modeling and preventing progressive hearing loss in Usher syndrome III. Sci Rep 7:13480

    Article  PubMed  PubMed Central  Google Scholar 

  22. Isgrig K, Shteamer JW, Belyantseva IA, Drummond MC, Fitzgerald TS, Vijayakumar S, Jones SM, Griffith AJ, Friedman TB, Cunningham LL, Chien WW (2017) Gene therapy restores balance and auditory functions in a mouse model of Usher syndrome. Mol Ther 25(3):780–791

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Pan B, Askew C, Galvin A, Heman-Ackah S, Asai Y, Indzhykulian AA, Jodelka FM, Hastings ML, Lentz JJ, Vandenberghe LH, Holt JR, Géléoc GS (2017) Gene therapy restores auditory and vestibular function in a mouse model of Usher syndrome type 1c. Nat Biotechnol 35(3):264–272

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Suzuki J, Hashimoto K, Xiao R, Vandenberghe LH, Liberman MC (2017) Cochlear gene therapy with ancestral AAV in adult mice: complete transduction of inner hair cells without cochlear dysfunction. Sci Rep 7:45524

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Akil O, Lustig LR (2013) Mouse cochlear whole mount immunofluorescence. Bio Protoc 3(5):e332

    Article  PubMed  PubMed Central  Google Scholar 

  26. Akil O, Chang J, Hiel H, Kong JH, Yi E, Glowatzki E, Lustig LR (2006) Progressive deafness and altered cochlear innervation in knock-out mice lacking prosaposin. J Neurosci 26(5):13076–13088

    Article  CAS  PubMed  Google Scholar 

  27. Fremeau RT, Kam K, Qureshi T, Johnson J, Copenhagen DR, Storm-Mathisen J, Chaudhry FA, Nicoll RA, Edwards RH (2004) Vesicular glutamate transporters 1 and 2 target to functionally distinct synaptic release sites. Science 304(5678):1815–1819

    Article  CAS  PubMed  Google Scholar 

  28. Akil O, Oursler AE, Fan K, Lustig LR (2016) Mouse auditory brainstem response testing. Bio Protoc 6(6):e1768

    Article  PubMed  PubMed Central  Google Scholar 

  29. Liu Y, Okada T, Sheykholeslami K, Shimazaki K, Nomoto T, Muramatsu S, Kanazawa T, Takeuchi K, Ajalli R, Mizukami H, Kume A, Ichimura K, Ozawa K (2005) Specific and efficient transduction of cochlear inner hair cells with recombinant adeno-associated virus type 3 vector. Mol Ther 12 (4:725–733

    Article  Google Scholar 

Download references

Acknowledgments

The authors would like to acknowledge the financial support provided by Hearing Research Inc. (HRI). We thank Neuron for some illustrating images we used in this chapter.

Author information

Authors and Affiliations

  1. Department of Otolaryngology-Head and Neck Surgery, University of California San Francisco, San Francisco, CA, USA

    Omar Akil

  2. Otolaryngology-Head and Neck Surgery, Columbia University Medical Center, New York, NY, USA

    Lawrence Lustig

Authors
  1. Omar Akil
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lawrence Lustig
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lawrence Lustig .

Editor information

Editors and Affiliations

  1. Department of Neurosciences, University of California, San Diego, La Jolla, CA, USA

    Michael J. Castle

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Akil, O., Lustig, L. (2019). AAV-Mediated Gene Delivery to the Inner Ear. In: Castle, M. (eds) Adeno-Associated Virus Vectors. Methods in Molecular Biology, vol 1950. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9139-6_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9139-6_16

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9138-9

  • Online ISBN: 978-1-4939-9139-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation