Adenovector-mediated hair cell regeneration is affected by  promoter type.
CONCLUSIONS
Replacement of vestibular hair cells induced by atoh1 driven by the tissue-specific GFAP promoter was significantly more efficient than use of the cBA or hCMV promoter.


OBJECTIVE
To test whether expression level, persistence, or selectivity from adenovirus vectors delivered in the inner ear can be altered by changing the adenovector backbone or by using different cellular and viral promoters.


METHODS
Adenovector and promoter modifications were tested for differences in transgene expression in adult macular organs. The effect of using an E1/E3 deleted vector was compared to E1/E3/E4 deleted vectors. The effect of using viral and cellular promoters to modify transgene expression was tested in explanted adult mouse macular organs. Based on these results three different promoters were tested for efficacy of atonal gene.


RESULTS
Use of adenovectors containing human CMV, the hybrid cBA and ubiquitin promoters driving transgene expression resulted in different types of transgene expression. While several viral and cellular promoters provided broad cell type expression, expression driven by the GFAP promoter was limited to vestibular supporting cells, demonstrating the specificity of this promoter.
Praetorius
Mark
M
Department of Otolaryngology, University of Heidelberg, Heidelberg, Germany.
Hsu
Chi
C
Baker
Kim
K
Brough
Douglas E
DE
Plinkert
Peter
P
Staecker
Hinrich
H
D008267:Acoustic Maculae / Q000502:physiology; D017500:Adenovirus E1A Proteins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics* / Q000378:metabolism; D004195:Disease Models, Animal; D007758:Ear, Inner / Q000502:physiology; D015967:Gene Expression Regulation, Viral / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018069:Hair Cells, Vestibular / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D011401:Promoter Regions, Genetic / Q000235:genetics; D012038:Regeneration / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic; D019076:Transgenes / Q000235:genetics; D014452:Ubiquitins / Q000235:genetics / Q000378:metabolism
D017500:Adenovirus E1A Proteins; D005904:Glial Fibrillary Acidic Protein; D014452:Ubiquitins; D049452:Green Fluorescent Proteins