Breidenbach M, Rein D T, Everts M, Glasgow J N, Wang M, Passineau M J, Alvarez R D, Korokhov N, Curiel D T
Division of Human Gene Therapy, Departments of Medicine, Surgery, Pathology and the Gene Therapy Center, Birmingham, AL 35294-2172, USA.
Gene Ther. 2005 Jan;12(2):187-93. doi: 10.1038/sj.gt.3302404.
Adenoviruses (Ads) are efficient gene transfer vehicles, but Ad-mediated gene therapy for ovarian cancer remains limited in vivo by inefficient and nonspecific gene transfer. Mesothelin (MSLN), a cell surface glycoprotein, is overexpressed in ovarian cancer but not in normal tissues except mesothelial cells. Therefore, MSLN is an attractive candidate for transcriptional and transductional targeting in the context of ovarian cancer gene therapy. We evaluated the expression of MSLN mRNA and MSLN surface protein in ovarian cancer cells. Ads containing the MSLN promoter driving reporter gene expression were created and tested in ovarian cancer cell lines and purified ovarian cancer cells isolated from patients. To evaluate transductional targeting, we used an Ad vector containing an Fc-binding domain within the fiber protein, which served as a docking domain for binding with anti-MSLN immunoglobulins. Both RT-PCR and flow cytometry revealed high MSLN gene and protein expression in ovarian cancer cells. The MSLN promoter was activated in ovarian cancer cells, but showed significantly reduced activity in normal control cells. Transductional targeting of Ads via anti-MSLN antibody increased transgene expression in ovarian cancer cells. This report describes the use of MSLN for transcriptional as well as transductional targeting strategies for ovarian cancer gene therapy.
腺病毒（Ads）是高效的基因传递载体，但腺病毒介导的卵巢癌基因治疗在体内仍受限于低效和非特异性的基因传递。间皮素（MSLN）是一种细胞表面糖蛋白，在卵巢癌中过表达，而在除间皮细胞外的正常组织中不表达。因此，在卵巢癌基因治疗中，MSLN是转录和转导靶向的一个有吸引力的候选分子。我们评估了卵巢癌细胞中MSLN mRNA和MSLN表面蛋白的表达。构建了含有驱动报告基因表达的MSLN启动子的腺病毒，并在卵巢癌细胞系和从患者分离的纯化卵巢癌细胞中进行了测试。为了评估转导靶向，我们使用了一种在纤维蛋白内含有Fc结合域的腺病毒载体，该结构域作为与抗MSLN免疫球蛋白结合的对接域。逆转录聚合酶链反应（RT-PCR）和流式细胞术均显示卵巢癌细胞中MSLN基因和蛋白高表达。MSLN启动子在卵巢癌细胞中被激活，但在正常对照细胞中活性显著降低。通过抗MSLN抗体对腺病毒进行转导靶向可增加卵巢癌细胞中的转基因表达。本报告描述了MSLN在卵巢癌基因治疗的转录和转导靶向策略中的应用。
