Barker S D, Dmitriev I P, Nettelbeck D M, Liu B, Rivera A A, Alvarez R D, Curiel D T, Hemminki A
Division of Human Gene Therapy, Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA.
Gene Ther. 2003 Jul;10(14):1198-204. doi: 10.1038/sj.gt.3301974.
Adenoviruses are efficient gene delivery vehicles but have broad native tropism. To this end, finding ways to target this virus specifically to carcinomas has become an important focus of cancer gene therapy. Transductional and transcriptional forms of targeting have been used with promising results in ovarian carcinoma. Therefore, we combined both forms of targeting to investigate the effect on the specificity and efficiency of transgene expression in this disease. We used the tissue-specific SLPI promoter and the ovarian cancer associated targeting adaptor protein, sCARfC6.5. This bispecific protein contains the coxsackie-adenovirus receptor ectodomain and a single-chain antibody specific for c-erbB-2. Viruses containing the SLPI or the ubiquitously expressed CMV promoter, with or without sCARfC6.5, were used for infection of ovarian cancer cell lines, primary ovarian tumor cells, and in an orthotopic model of disseminated ovarian carcinoma. This dual-targeting strategy increased the efficiency and specificity of transgene expression in vitro in reporter and cell-killing assays, and in vivo. By using both the SLPI promoter and sCARfC6.5, transgene expression was increased in ovarian tumors and decreased in normal tissues, including the liver. Thus, we show that combining transcriptional and transductional targeting can increase the efficacy and specificity of adenoviral gene therapy for ovarian carcinoma.
腺病毒是高效的基因传递载体，但具有广泛的天然嗜性。为此，找到将这种病毒特异性靶向癌组织的方法已成为癌症基因治疗的一个重要焦点。在卵巢癌中，转导靶向和转录靶向形式都已被使用并取得了有前景的结果。因此，我们将这两种靶向形式结合起来，以研究其对该疾病中转基因表达的特异性和效率的影响。我们使用了组织特异性的鳞状上皮细胞癌抗原启动子（SLPI启动子）和卵巢癌相关的靶向衔接蛋白sCARfC6.5。这种双特异性蛋白包含柯萨奇病毒-腺病毒受体胞外域和对c-erbB-2特异的单链抗体。含有SLPI或普遍表达的巨细胞病毒（CMV）启动子、有或没有sCARfC6.5的病毒，被用于感染卵巢癌细胞系、原发性卵巢肿瘤细胞，以及在播散性卵巢癌的原位模型中。这种双靶向策略在体外的报告基因和细胞杀伤试验以及体内均提高了转基因表达的效率和特异性。通过同时使用SLPI启动子和sCARfC6.5，转基因表达在卵巢肿瘤中增加，而在包括肝脏在内的正常组织中减少。因此，我们表明，将转录靶向和转导靶向相结合可以提高腺病毒基因治疗卵巢癌的疗效和特异性。
