Barker Shannon D, Coolidge Candace J, Kanerva Anna, Hakkarainen Tanja, Yamamoto Masato, Liu Bin, Rivera Angel A, Bhoola Snehal M, Barnes Mack N, Alvarez Ronald D, Curiel David T, Hemminki Akseli
Division of Human Gene Therapy, Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
J Gene Med. 2003 Apr;5(4):300-10. doi: 10.1002/jgm.341.
Adenoviruses allow efficient transduction of dividing and non-dividing cells and their safety for the treatment of cancer has been established in clinical trials. However, one disadvantage is their promiscuous tropism. In this regard, tissue-specific promoters (TSPs) could be useful for directing transgene expression to target tissues and for reducing adverse effects in non-target tissues. We hypothesize that selective adenovirus-mediated transgene expression could be achieved through the use of the secretory leukoprotease inhibitor (SLPI) promoter in the context of ovarian cancer.
Adenoviruses containing the SLPI promoter driving reporter and suicide gene expression were created and tested in ovarian cancer cell lines and primary tumor cells isolated from patients. To evaluate the in vivo activation of the SLPI promoter in comparison to a ubiquitous promoter, intraperitoneal delivery was performed in tumor-bearing mice, followed by analysis of survival or gene expression in normal organs and tumor.
The SLPI promoter retained its fidelity in an adenoviral context and was activated in both cell lines and primary cancer cells. The SLPI promoter was induced to a high degree in ovarian cancer cells while showing significantly reduced activity in normal tissues. The therapeutic efficacy of SLPI promoter-controlled gene expression was similar to the ubiquitous promoter in vitro and in an orthotopic murine model of peritoneally disseminated ovarian cancer, with higher activity than controls.
The SLPI promoter is a potentially useful TSP for ovarian cancer and facilitates further development of targeting strategies for improved gene therapy of ovarian carcinomas.
腺病毒可有效转导分裂细胞和非分裂细胞，其用于癌症治疗的安全性已在临床试验中得到证实。然而，其一个缺点是具有广泛的嗜性。在这方面，组织特异性启动子（TSPs）可用于将转基因表达导向靶组织，并减少非靶组织中的不良反应。我们假设在卵巢癌的背景下，通过使用分泌型白细胞蛋白酶抑制剂（SLPI）启动子可实现选择性腺病毒介导的转基因表达。
构建了含有驱动报告基因和自杀基因表达的SLPI启动子的腺病毒，并在卵巢癌细胞系和从患者分离的原发性肿瘤细胞中进行了测试。为了评估与普遍存在的启动子相比SLPI启动子在体内的激活情况，对荷瘤小鼠进行腹腔注射，随后分析正常器官和肿瘤中的存活率或基因表达。
SLPI启动子在腺病毒环境中保持其保真度，并在细胞系和原发性癌细胞中均被激活。SLPI启动子在卵巢癌细胞中被高度诱导，而在正常组织中活性显著降低。在体外和腹膜播散性卵巢癌的原位小鼠模型中，SLPI启动子控制的基因表达的治疗效果与普遍存在的启动子相似，且活性高于对照。
SLPI启动子是一种对卵巢癌潜在有用的TSP，有助于进一步开发靶向策略，以改善卵巢癌的基因治疗。
