Specific and efficient transduction of Cochlear inner hair cells with recombinant adeno-associated virus type 3 vector.
Recombinant adeno-associated virus (AAV) vectors are of interest for cochlear gene therapy because of their ability to mediate the efficient transfer and long-term stable expression of therapeutic genes in a wide variety of postmitotic tissues with minimal vector-related cytotoxicity. In the present study, seven AAV serotypes (AAV1-5, 7, 8) were used to construct vectors. The expression of EGFP by the chicken beta-actin promoter associated with the cytomegalovirus immediate-early enhancer in cochlear cells showed that each of these serotypes successfully targets distinct cochlear cell types. In contrast to the other serotypes, the AAV3 vector specifically transduced cochlear inner hair cells with high efficiency in vivo, while the AAV1, 2, 5, 7, and 8 vectors also transduced these and other cell types, including spiral ganglion and spiral ligament cells. There was no loss of cochlear function with respect to evoked auditory brain-stem responses over the range of frequencies tested after the injection of AAV vectors. These findings are of value for further molecular studies of cochlear inner hair cells and for gene replacement strategies to correct recessive genetic hearing loss due to monogenic mutations in these cells.
Liu
Yuhe
Y
Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School, Minami-kawachi, Kawachi, Tochigi, Japan.
Okada
Takashi
T
Sheykholeslami
Kianoush
K
Shimazaki
Kuniko
K
Nomoto
Tatsuya
T
Muramatsu
Shin-Ichi
S
Kanazawa
Takeharu
T
Takeuchi
Koichi
K
Ajalli
Rahim
R
Mizukami
Hiroaki
H
Kume
Akihiro
A
Ichimura
Keiichi
K
Ozawa
Keiya
K
D017498:Adenovirus E1 Proteins; D000818:Animals; D003051:Cochlea / Q000502:physiology; D000229:Dependovirus / Q000145:classification / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D006199:Hair Cells, Auditory, Inner; D034381:Hearing Loss; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D008856:Microscopy, Fluorescence; D010957:Plasmids; D012703:Serotyping; D014161:Transduction, Genetic; D019076:Transgenes
D017498:Adenovirus E1 Proteins; D049452:Green Fluorescent Proteins