title: Muscle derived cell mediated ex vivo gene transfer to the lower urinary tract: comparison of viral vectors.
abstract: Gene therapy is a novel form of molecular medicine that may have a major impact on the future of human health care. We explored the efficacy of skeletal muscle derived cells (MDC) transduced with four viruses for ex vivo gene transfer into the lower urinary tract. Primary MDC were isolated from normal neonatal rats and transduced with: (1). adenovirus, (2). herpes simplex virus type-1 (HSV-1), (3). retrovirus or (4). adeno-associated virus (AAV), all of which express the beta-galactosidase reporter gene. Adult Sprague Dawley rats ( n=4 each group-time) were used. The MDC were injected into the right and left lateral bladder walls. The number of injected MDC ranged from 1 to 1.5 x 10(6). The tissues were harvested after 1, 4, 7, and 15 days, sectioned and assayed for beta-galactosidase expression. In the bladder wall, we noted cells expressing beta-galactosidase for each viral group. Adenoviral and HSV-1 transduced cells showed strong expression at 1 and 4 days post-injection, but the expression decreased gradually and was not detectable at 15 days post-injection. Retroviral transduced cells were detected at each time point with a strong expression persisting for 15 days but decreasing gradually over time. Although expression of the AAV transduced cells was initially weak, the later time points exhibited a much stronger expression, especially at day 7 post-injection. This expression persisted for at least 15 days post-injection. In conclusion, successful MDC mediated ex vivo gene transfer into the lower urinary tract was achieved with all four viral vectors. Our results suggest that the ex vivo approach may lead to an efficient and persistent viral gene delivery to the lower urinary tract while minimizing exposure of the host to virus.
authors.affiliation: Department of Orthopedic Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Pruchnic
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Yokoyama
authors.forename: Ji Youl
authors.initials: JY
authors.lastname: Lee
authors.forename: Johnny
authors.initials: J
authors.lastname: Huard
authors.forename: Michael B
authors.initials: MB
authors.lastname: Chancellor
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017690:Cell Transplantation; D000229:Dependovirus / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics; D007267:Injections; D018482:Muscle, Skeletal / Q000166:cytology* / Q000821:virology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012190:Retroviridae / Q000235:genetics; D013997:Time Factors; D014551:Urinary Tract; D014553:Urinary Tract Physiological Phenomena; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D001616:beta-Galactosidase