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 vi...

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Bibliographic Details
Published in:Human reproduction (Oxford) 2008-03, Vol.23 (3), p.514-524
Main Authors: Hassan, M.H., Khatoon, N., Curiel, D.T., Hamada, F.M., Arafa, H.M., Al-Hendy, A.
Format: Article
Language:English
Subjects:
Adenoviridae - genetics
Adenovirus
adenovirus targeting strategies
Biological and medical sciences
Female
gene therapy
Genetic Therapy - methods
Gynecology. Andrology. Obstetrics
Humans
Leiomyoma - therapy
Leiomyoma - virology
Liver - cytology
Medical sciences
Myometrium - cytology
Myometrium - virology
Receptors, Virus - genetics
uterine leiomyoma
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Summary: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.
ISSN:0268-1161
1460-2350
DOI:10.1093/humrep/dem410