Coxsackie adenovirus receptor and alpha nu beta3/alpha nu beta5 integrins in adenovirus gene transfer of rat cochlea.
This study was designed to determine whether Coxsackie adenovirus receptor (CAR) and alpha nu beta3/alpha nu beta5 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 DeltaE1-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 alpha nu 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-alpha nu antibodies decreases GFP expression in vivo, suggesting that the CAR/alpha nu integrin pathway is involved in adenovirus transduction in the cochlea.
Venail
F
F
INSERM UMR 583, Physiopathologie et Thérapie des Déficits Sensoriels et Moteurs, Montpellier, France.
Wang
J
J
Ruel
J
J
Ballana
E
E
Rebillard
G
G
Eybalin
M
M
Arbones
M
M
Bosch
A
A
Puel
J-L
JL
D000200:Action Potentials; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D003051:Cochlea / Q000378:metabolism* / Q000821:virology; D003056:Cochlear Nerve / Q000502:physiology; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D007267:Injections; D039482:Integrin alpha5 / Q000032:analysis / Q000378:metabolism; D039641:Integrin beta Chains / Q000032:analysis / Q000378:metabolism; D039661:Integrin beta3 / Q000032:analysis / Q000378:metabolism; D016023:Integrins / Q000032:analysis / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D023421:Models, Animal; D051381:Rats; D017208:Rats, Wistar; D011991:Receptors, Virus / Q000378:metabolism*; D046509:Tissue Culture Techniques; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes
C568771:CLMP protein, human; C568773:Clmp protein, rat; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D039482:Integrin alpha5; D039641:Integrin beta Chains; D039661:Integrin beta3; D016023:Integrins; D011991:Receptors, Virus; C084344:integrin beta5; D049452:Green Fluorescent Proteins