A modified adenovirus can transfect cochlear hair cells in vivo without compromising cochlear function

Luebke, Anne E.; Steiger, Julia; Hodges, Bradley L.; Amalfitano, Andrea

doi:10.1038/sj.gt.3301445

Cited by 76 publications

(65 citation statements)

References 28 publications

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“…Indeed, gene expression in the sensory hair cells and strial cells has previously been reported using the same syringe inoculation protocol. 10 Intriguing, however, are the results of Luebke et al, 8,9 who reported that 7 days of delivery of adenovirus into the basal turn perilymph via an osmotic minipump led to preservation of hearing and b-gal expression specifically in the sensory hair cells, but not into the adjacent structures such as the supporting cells. In Accordance with Ishimoto et al, 6 who was unable to replicate this result, we suggest that the method used to detect b-gal expression may account for this unexpected result.…”

Section: Efficiency and Safety Of Gene Transfer In Vivomentioning

confidence: 65%

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Coxsackie adenovirus receptor and ανβ3/ανβ5 integrins in adenovirus gene transfer of rat cochlea

et al. 2006

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This study was designed to determine whether Coxsackie adenovirus receptor (CAR) and anb3/anb5 integrin co-receptors are involved in adenovirus gene transfer in the rat cochlea. We find that CAR and integrin co-receptors are expressed in every cell subtype transduced by the adenoviral vector Ad5 DE1-E3/cytomegalovirus/green fluorescent protein (GFP) on cochlear slices in vitro. The spiral ganglion neurons, which do not express CAR, were not transduced by the virus. Blocking these receptors by monoclonal antibodies decreased transgene expression, whereas disrupting tight junctions with ethylenediaminetetraacetic acid led to an increased transgene expression. However, sensory hair cells and strial cells also expressing CAR and an integrins were not transduced by the vector.GFP expression was also studied in vivo. Perilymphatic perfusion of adenovirus in vivo did not affect hearing and only cells lining the perilymphatic spaces were transduced. Endolymphatic perfusion resulted in low-frequency hearing loss and although some cells of the organ of Corti were efficiently transduced, the sensory and the strial cells were not. Transduced sensory and strial cells were occasionally observed in cochleas after single shot of adenovirus. Pretreatment with anti-CAR and anti-an antibodies decreases GFP expression in vivo, suggesting that the CAR/an integrin pathway is involved in adenovirus transduction in the cochlea.

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Section: Efficiency and Safety Of Gene Transfer In Vivomentioning

confidence: 65%

“…6,7 However, other studies have demonstrated that adenovirus vectors can indeed transduce sensory hair cells in the organ of Corti. [8][9][10] So far, no explanation has been provided to explain this discrepancy.…”

Section: Introductionmentioning

confidence: 99%

Coxsackie adenovirus receptor and ανβ3/ανβ5 integrins in adenovirus gene transfer of rat cochlea

et al. 2006

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“…Agents have been identified that can minimize degeneration and facilitate repair [4,[8][9][10][11][12]14]. Moreover, the extraordinary progress that has been made in defining the genes involved in a number of human genetic forms of deafness [15,16] offers hope for gene-transfer [1,17] and molecular [18] approaches to treat these diseases.…”

Section: Introductionmentioning

confidence: 99%

Inner ear drug delivery via a reciprocating perfusion system in the guinea pig

Chen

Kujawa

McKenna

et al. 2005

Journal of Controlled Release

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Rapid progress in understanding the molecular mechanisms associated with cochlear and auditory nerve degenerative processes offers hope for the development of gene-transfer and molecular approaches to treat these diseases in patients. For therapies based on these discoveries to become clinically useful, it will be necessary to develop safe and reliable mechanisms for the delivery of drugs into the inner ear, bypassing the blood-labyrinthine barrier. Toward the goal of developing an inner ear perfusion device for human use, a reciprocating microfluidic system that allows perfusion of drugs into the cochlear perilymph through a single inlet hole in scala tympani of the basal turn was developed. The performance of a prototype, extracorporeal reciprocating perfusion system in guinea pigs is described. Analysis of the cochlear distribution of compounds after perfusion took advantage of the place-dependent generation of responses to tones along the length of the cochlea. Perfusion with a control artificial perilymph solution had no effect. Two drugs with wellcharacterized effects on cochlear physiology, salicylate (5 mM) and DNQX (6,7-Dinitroquinoxaline-2,3-dione; 100 and 300 μM), reversibly altered responses. The magnitude of drug effect decreased with distance from the perfusion pipette for up to 10 mm, and increased with dose and length of application.

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“…We tested vectors with a single gene deleted and with multiple genes deleted and found that both vector designs transfected sensory epithelial cells. Previously, E1a/b-deleted vectors were shown to be toxic to mouse vestibular hair cells, 5,6 but multiply deleted adenoviral vectors were found to have reduced or no hair cell toxicity. [3][4][5] We did not detect any direct toxic effects on the cultured cells for either the E1a/b-deleted or the multiply deleted adenoviral vectors, but chose to focus our work on multiply deleted vectors since these would most likely have reduced toxicity in vivo.…”

Section: Cell Viability and Availabilitymentioning

confidence: 99%

“…Previously, E1a/b-deleted vectors were shown to be toxic to mouse vestibular hair cells, 5,6 but multiply deleted adenoviral vectors were found to have reduced or no hair cell toxicity. [3][4][5] We did not detect any direct toxic effects on the cultured cells for either the E1a/b-deleted or the multiply deleted adenoviral vectors, but chose to focus our work on multiply deleted vectors since these would most likely have reduced toxicity in vivo. Although cellular morphology of transfected cells appeared similar to that of neighboring nontransfected cells and transfected cells were able to synthesize the exogenous protein and remain viable for up to 4 days, studies in which cell physiology, including measurement of mechanotransduction in human hair cells before and after viral transfection, will be required to more directly address vector toxicity.…”

Section: Cell Viability and Availabilitymentioning

confidence: 99%

An in vitro model system to study gene therapy in the human inner ear

et al. 2007

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The confined fluid-filled labyrinth of the human inner ear presents an opportunity for introduction of gene therapy reagents designed to treat hearing and balance dysfunction. Here we present a novel model system derived from the sensory epithelia of human vestibular organs and show that the tissue can survive up to 5 days in vitro. We generated organotypic cultures from 26 human sensory epithelia excised at the time of labyrinthectomy for intractable Meniere's disease or vestibular schwannoma. We applied multiply deleted adenoviral vectors at titers between 10 5 and 10 8 viral particles/ml directly to the cultures for 4-24 h and examined the tissue 12-96 h post-transfection. We noted robust expression of the exogenous transgene, green fluorescent protein (GFP), in hair cells and supporting cells suggesting both were targets of adenoviral transfection. We also transfected cultures with a vector that carried the genes for GFP and KCNQ4, a potassium channel subunit that causes dominant-progressive hearing loss when mutated. We noted a positive correlation between GFP fluorescence and KCNQ4 immunolocalization. We conclude that our in vitro model system presents a novel and effective experimental paradigm for evaluation of gene therapy reagents designed to restore cellular function in patients who suffer from inner ear disorders.

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A modified adenovirus can transfect cochlear hair cells in vivo without compromising cochlear function

Cited by 76 publications

References 28 publications

Coxsackie adenovirus receptor and ανβ3/ανβ5 integrins in adenovirus gene transfer of rat cochlea

Coxsackie adenovirus receptor and ανβ3/ανβ5 integrins in adenovirus gene transfer of rat cochlea

Inner ear drug delivery via a reciprocating perfusion system in the guinea pig

An in vitro model system to study gene therapy in the human inner ear

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