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Gene transfer to urethral strictures in rabbits: a preliminary report
Author(s) -
Meria P.,
Anidjar M.,
Brouland J.P.,
Teillac P.,
Berthon P.,
Cussenot O.
Publication year - 2000
Publication title -
bju international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.773
H-Index - 148
eISSN - 1464-410X
pISSN - 1464-4096
DOI - 10.1046/j.1464-410x.2000.00615.x
Subject(s) - urethra , stromal cell , retrovirus , reporter gene , transduction (biophysics) , genetic enhancement , immunohistochemistry , transfection , biology , pathology , microbiology and biotechnology , cell culture , medicine , virus , gene , virology , gene expression , anatomy , biochemistry , genetics
Objective To assess the rationale for virus‐mediated gene transfer into the urethra in vivo and in vitro , using a rabbit model, as this is an attractive approach to prevent recurrence after the endoscopic management of urethral strictures. Materials and methods Primary cultures of rabbit urethral stromal cells were infected with adenoviral and retroviral solutions carrying a nucleus‐targeted β‐galactosidase (β‐Gal) reporter gene (respectively 10 9 and 10 7 plaque‐forming units/mL). In addition, to mimic the human clinical situation, a model was developed of thermally induced stricture in rabbit urethra which produced fibrotic stenosis within 15 days. Using a prototype channelled balloon catheter, these strictures were endoscopically dilated and then instilled with the β‐Gal adenoviral or retroviral constructs. Results The application of recombinant adenovirus and retrovirus harbouring a nucleus‐targeted β‐Gal reporter gene to cultured rabbit urethral stromal cells resulted in a high transduction efficiency of up to 90% and 96%, respectively. Five days after infection, histochemical and immunohistochemical staining of the strictured urethrae showed a 3% rate of transfection targeted to stromal cells within the fibrosis, confirmed by polymerase chain reaction (PCR) analysis. Adjacent and distal spread of the virus was excluded by histochemistry, immunohistochemistry and PCR. Conclusion These results represent the first report of endoscopic adenovirus and retrovirus‐mediated gene transfer to the urethra. Although at a low rate, transduction reached stromal cells transmurally within the induced strictures and was site‐specific.