title: Viral vector tropism for supporting cells in the developing murine cochlea.
abstract: Gene-based therapeutics are being developed as novel treatments for genetic hearing loss. One roadblock to effective gene therapy is the identification of vectors which will safely deliver therapeutics to targeted cells. The cellular heterogeneity that exists within the cochlea makes viral tropism a vital consideration for effective inner ear gene therapy. There are compelling reasons to identify a viral vector with tropism for organ of Corti supporting cells. Supporting cells are the primary expression site of connexin 26 gap junction proteins that are mutated in the most common form of congenital genetic deafness (DFNB1). Supporting cells are also primary targets for inducing hair cell regeneration. Since many genetic forms of deafness are congenital it is necessary to administer gene transfer-based therapeutics prior to the onset of significant hearing loss. We have used transuterine microinjection of the fetal murine otocyst to investigate viral tropism in the developing inner ear. For the first time we have characterized viral tropism for supporting cells following in utero delivery to their progenitors. We report the inner ear tropism and potential ototoxicity of three previously untested vectors: early-generation adenovirus (Ad5.CMV.GFP), advanced-generation adenovirus (Adf.11D) and bovine adeno-associated virus (BAAV.CMV.GFP). Adenovirus showed robust tropism for organ of Corti supporting cells throughout the cochlea but induced increased ABR thresholds indicating ototoxicity. BAAV also showed tropism for organ of Corti supporting cells, with preferential transduction toward the cochlear apex. Additionally, BAAV readily transduced spiral ganglion neurons. Importantly, the BAAV-injected ears exhibited normal hearing at 5 weeks of age when compared to non-injected ears. Our results support the use of BAAV for safe and efficient targeting of supporting cell progenitors in the developing murine inner ear.
authors.affiliation: Department of Otolaryngology, University of Iowa, Iowa City, IA, USA.
authors.forename: Abraham M
authors.initials: AM
authors.lastname: Sheffield
authors.forename: Samuel P
authors.initials: SP
authors.lastname: Gubbels
authors.forename: Michael S
authors.initials: MS
authors.lastname: Hildebrand
authors.forename: Stephen S
authors.initials: SS
authors.lastname: Newton
authors.forename: John A
authors.initials: JA
authors.lastname: Chiorini
authors.forename: Giovanni
authors.initials: G
authors.lastname: Di Pasquale
authors.forename: Richard J H
authors.initials: RJ
authors.lastname: Smith
mesh_terms: D000161:Acoustic Stimulation; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001301:Audiometry, Pure-Tone; D001309:Auditory Threshold; D003638:Deafness / Q000235:genetics / Q000503:physiopathology / Q000628:therapy*; D000229:Dependovirus / Q000235:genetics*; D016057:Evoked Potentials, Auditory, Brain Stem; D018014:Gene Transfer Techniques / Q000009:adverse effects; D015316:Genetic Therapy / Q000009:adverse effects / Q000379:methods*; D005822:Genetic Vectors / Q000009:adverse effects; D005865:Gestational Age; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D007760:Labyrinth Supporting Cells / Q000378:metabolism / Q000821:virology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008845:Microinjections; D008856:Microscopy, Fluorescence; D009925:Organ of Corti / Q000196:embryology / Q000378:metabolism / Q000503:physiopathology / Q000821:virology*; D013234:Stem Cells / Q000821:virology; D014161:Transduction, Genetic; D056189:Viral Tropism
chemical_list: D049452:Green Fluorescent Proteins