Toward gene therapy of uterine fibroids: targeting modified adenovirus to human leiomyoma cells

被引:17
作者
Hassan, M. H. [3 ,4 ]
Khatoon, N. [4 ]
Curiel, D. T. [2 ]
Hamada, F. M. [3 ]
Arafa, H. M. [3 ]
Al-Hendy, A. [1 ]
机构
[1] Meharry Med Coll, Ctr Womens Hlth Res, Dept Obstet & Gynecol, Nashville, TN 37208 USA
[2] Univ Alabama Birmingham, Gene Therapy Ctr, Birmingham, AL 35924 USA
[3] Al Azhar Univ, Dept Pharmacol & Toxicol, Fac Pharm, Cairo, Egypt
[4] Univ Texas Galveston, Med Branch, Dept Obstet & Gynecol, Galveston, TX 77555 USA
关键词
uterine leiomyoma; gene therapy; adenovirus targeting strategies;
D O I
10.1093/humrep/dem410
中图分类号
R71 [妇产科学];
学科分类号
100211 ;
摘要
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.
引用
收藏
页码:514 / 524
页数:11
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