Towards gene therapy of postoperative adhesions: fiber and transcriptional modifications enhance adenovirus targeting towards human adhesion cells.
Postoperative abdominal/pelvic peritoneal adhesions are a major source of morbidity (bowel obstruction, infertility, ectopic gestation as well as chronic pelvic pain) in women. In this study, we screened various transduction and transcription modifications of adenovirus (Ad) to identify those that support maximal Ad-mediated gene delivery to human adhesion fibroblasts, which in turn would enhance the efficacy of this novel treatment/preventative strategy for postoperative adhesions. We transduced primary cultures of human peritoneal adhesion fibroblasts with fiber-modified Ad vectors Ad5-RGD-luc, Ad5-Sigma-luc, Ad5/3-luc and Ad5-CAV2-luc as well as transcriptional targeting viruses Ad5-survivin-luc, Ad5-heparanase-luc, Ad5-mesothelin (MSLN)-CRAd-luc and Ad5-secretory leukoprotease inhibitor (SLPI)-luc, and compared their activity to wild-type Ad5-luc. At 48 h, luciferase activity was measured and normalized to the total protein content in the cells. Among the fiber-modified Ad vectors, Ad5-Sigma-luc and among the transcriptional targeting modified Ad vectors, Ad5-MSLN-CRAd-luc showed significantly increased expression levels of luciferase activity at 5, 10 and 50 plaque forming units/cell in adhesion fibroblast cells compared with wild-type Ad5-luc (p < 0.05). Specific modifications of Ad improve their gene delivery efficiency towards human peritoneal adhesion fibroblasts. Developing a safe localized method to prevent/treat postoperative adhesion formation would have a major impact on women health.
Center for Women's Health Research, Department of Obstetrics and Gynecology, Meharry Medical College, Nashville, Tenn., USA.
S
S
Nair
G M
GM
Saed
H M
HM
Atta
V
V
Rajaratnam
M P
MP
Diamond
D T
DT
Curiel
A
A
Al-Hendy
D000256:Adenoviridae / Q000235:genetics*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000201:enzymology; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D000090204:Mesothelin; D000267:Tissue Adhesions / Q000517:prevention & control / Q000628:therapy*; D014158:Transcription, Genetic; D014161:Transduction, Genetic
C000715527:MSLN protein, human; D008156:Luciferases; D000090204:Mesothelin