Toward gene therapy of uterine fibroids: targeting modified adenovirus to human leiomyoma cells.
BACKGROUND
To circumvent the paucity of the primary adenovirus (Ad5) receptor and the non-specific Ad5 tropism in the context of uterine leiomyoma cells, Ad5 modification strategies would be beneficial.


METHODS
We screened several modified adenoviruses to identify the most efficient and selective virus toward human leiomyoma cells to be used as candidate for delivering therapeutic genes. We propagated: wild-type Ad5-luc, fiber-modified viruses: ad5 RGD-luc, Ad5-Sigma-luc, Ad5/3-luc and Ad5-CAV2-luc, as well as transcriptional targeted viruses: ad5 survivin-luc, Ad5-heparanase-luc, Ad5-MSLN-CRAD-luc and Ad5-SLPI-luc, on 293 cells and purified them by double CsCL density centrifugation. Then we transfected primary cultures of human leiomyoma cells derived from fibroids of four different patients, telomerase-immortalized human leiomyoma cell line (huLM), telomerase-immortalized normal human myometrial cell line (HM9) and immortalized normal human liver cells (THLE3) with the viruses at 5, 10 and 50 plaque-forming units (PFU)/cell. After 48 h, luciferase activities were measured and normalized to the total cellular protein content.


RESULTS
Ad5-RGD-luc and Ad5-CAV2-luc, Ad5-SLPI-luc and Ad5-MSLN-CRAD-luc at 5, 10 and 50 pfu/cell showed significantly higher expression levels of luciferase activity in both primary and immortalized human leiomyoma cells when compared with Ad5-Luc. Additionally, these modified viruses demonstrated selectivity toward leiomyoma cells, compared with myometrial cells and exhibited lower liver cell transduction, compared with Ad5-luc, at the same dose levels.


CONCLUSIONS
Ad5-CAV2-luc, Ad5-RGD-luc, Ad5-SLPI-luc and Ad5-MSLN-CRAD-luc are promising delivery vehicles in the context of leiomyoma gene therapy.
Hassan
M H
MH
Department of Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, TX 77555-0587, USA.
Khatoon
N
N
Curiel
D T
DT
Hamada
F M
FM
Arafa
H M
HM
Al-Hendy
A
A
D000256:Adenoviridae / Q000235:genetics*; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007889:Leiomyoma / Q000628:therapy* / Q000821:virology*; D008099:Liver / Q000166:cytology; D000090204:Mesothelin; D009215:Myometrium / Q000166:cytology / Q000821:virology; D011991:Receptors, Virus / Q000235:genetics
C000715527:MSLN protein, human; D011991:Receptors, Virus; C085301:adenovirus receptor; D000090204:Mesothelin