Transgene expression in the guinea pig cochlea mediated by a lentivirus-derived gene transfer vector.
The utility of lentivirus as a gene delivery vector in the cochlea was evaluated in vitro and in vivo. Lentivirus transduction was assessed through expression analysis of a reporter gene, green fluorescent protein (GFP), integrated within the viral genome. In vitro characterization of lentivirus-GFP was assessed by infection of explants from cochleas of neonatal rat. The lentiviral vector transduced both spiral ganglion neurons (SGNs) and glial cells. In vivo characterization of lentivirus-GFP was assessed by directly infusing the vector into the guinea pig cochlea via an osmotic minipump. Sections of lentivirus-infused cochlea revealed a highly restricted fluorescence pattern limited to the periphery of the perilymphatic space. Transduction of SGNs and glial cells by lentivirus in vitro but not in vivo suggests limited dissemination of the viral vector from the perilymphatic space. The cellular and tissue architecture of the lentivirus-infused cochlea was intact and free of inflammation. Restricted transduction of cell types confined to the periphery of the perilymphatic space by the lentivirus is ideal for stable production of gene products secreted into the perilymph.
Han
J J
JJ
Department of Otolaryngology--Head and Neck Surgery, University of California San Francisco, 94143, USA.
Mhatre
A N
AN
Wareing
M
M
Pettis
R
R
Gao
W Q
WQ
Zufferey
R N
RN
Trono
D
D
Lalwani
A K
AK
D000818:Animals; D002478:Cells, Cultured; D003051:Cochlea / Q000166:cytology / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006168:Guinea Pigs; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000821:virology; D009924:Organ Culture Techniques; D014161:Transduction, Genetic; D019076:Transgenes
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins