title: Optimizing atoh1-induced vestibular hair cell regeneration.
abstract: OBJECTIVE
  Determine the optimal design characteristics of an adenoviral (Ad) vector to deliver atoh1 and induce regeneration of vestibular hair cells.
  
  
  METHODS
  Evaluation of a mouse model of intralabyrinthine gene delivery. Tissue culture of mouse and human macular organs.
  
  
  METHODS
  Macular organs from adult C57Bl/6 mice were treated with binding modified and alternate adenovectors expressing green fluorescent protein (gfp) or luciferase (L). Expression of marker genes was determined over time to determine vector transfection efficiency. The inner ear of adult mice was then injected with modified vectors. Expression of gfp and distribution of vector DNA was followed. Hearing and balance function was evaluated in normal animals to ensure safety of the novel vector designs. An optimized vector was identified and tested for its ability to induce hair cell regeneration in a mouse vestibulopathy model. Finally, this vector was tested for its ability to induce hair cell regeneration in human tissue.
  
  
  RESULTS
  Ad5 serotype-based vectors were identified as having a variety of different binding capacities for inner ear tissue. This makes it difficult to limit the dose of vector due to entry into nontargeted cells. Screening of rare adenovector serotypes demonstrated that Ad-based vectors were ideally suited for delivery to supporting cells; therefore, they were useful for hair cell regeneration studies. Utilization of an Ad28-based vector to deliver atoh1 to a mouse model of vestibular loss resulted in significant functional recovery of balance. This vector was also capable of transfecting human macular organs and inducing regeneration of human vestibular hair cells in vitro.
  
  
  CONCLUSIONS
  Improvement in vector design can lead to more specific cell-based delivery and reduction of nonspecific delivery of the trans gene, leading to the development of optimized molecular therapeutics to induce hair cell regeneration.
  
  
  METHODS
  N/A. Laryngoscope 124:S1-S12, 2014.
authors.affiliation: Department of Otolaryngology, Head and Neck Surgery, University of Kansas School of Medicine, Kansas City, Kansas.
authors.forename: Hinrich
authors.initials: H
authors.lastname: Staecker
authors.forename: Christina
authors.initials: C
authors.lastname: Schlecker
authors.forename: Shannon
authors.initials: S
authors.lastname: Kraft
authors.forename: Mark
authors.initials: M
authors.lastname: Praetorius
authors.forename: Chi
authors.initials: C
authors.lastname: Hsu
authors.forename: Douglas E
authors.initials: DE
authors.lastname: Brough
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics*; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage*; D018069:Hair Cells, Vestibular / Q000502:physiology*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction / Q000379:methods; D004856:Postural Balance / Q000235:genetics; D011897:Random Allocation; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012038:Regeneration / Q000235:genetics*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; C500942:ATOH1 protein, human
keywords: Atoh1; adenovector; balance; hair cell regeneration; viral vector