Kuriyama N, Kuriyama H, Julin C M, Lamborn K, Israel M A
Preuss Laboratory for Molecular Neuro-oncology, Brain Tumor Research Center, Departments of Neurological Surgery and Pediatrics, University of California San Francisco, San Francisco, CA 94143, USA.
Hum Gene Ther. 2000 Nov 1;11(16):2219-30. doi: 10.1089/104303400750035744.
A key impediment to the development of effective virus-mediated gene therapy for cancer is the low level of gene transfer that occurs after the administration of recombinant viral vectors. Improving in vivo infection and transduction efficiency is an important goal for gene therapy. The limited distribution of gene delivery is particularly problematic when large vectors such as recombinant adenoviruses and retroviruses are used to mediate transgene delivery to solid tumors. To facilitate the spread of virus, we have investigated the potential of administering proteases prior to the intratumoral inoculation of recombinant replication deficient adenovirus. For these studies, we chose proteases that are active against collagen and the other extracellular matrix proteins found in primary brain tumor tissue, but are not widely expressed in normal brain. Various concentrations of a mixture of collagenase/dispase or trypsin were inoculated into xenografts of human glioblastoma multiforme-derived brain tumor cell lines U87, U251, and SF767. Subsequently, recombinant adenovirus encoding the beta-galactosidase gene was administered and tumor tissue was examined for evidence of virus infection. Both collagenase/dispase and trypsin enhanced virus infection, indicating that protease pretreatment may be a useful strategy for enhancing virus-mediated gene transduction for many in vivo applications.
开发有效的病毒介导癌症基因治疗的一个关键障碍是重组病毒载体给药后基因转移水平较低。提高体内感染和转导效率是基因治疗的一个重要目标。当使用重组腺病毒和逆转录病毒等大型载体介导转基因传递到实体瘤时，基因递送的有限分布尤其成问题。为了促进病毒传播，我们研究了在瘤内接种重组复制缺陷腺病毒之前施用蛋白酶的可能性。对于这些研究，我们选择了对原发性脑肿瘤组织中发现的胶原蛋白和其他细胞外基质蛋白有活性，但在正常脑中不广泛表达的蛋白酶。将各种浓度的胶原酶/分散酶或胰蛋白酶混合物接种到人多形性胶质母细胞瘤衍生的脑肿瘤细胞系U87、U251和SF767的异种移植瘤中。随后，施用编码β-半乳糖苷酶基因的重组腺病毒，并检查肿瘤组织是否有病毒感染的证据。胶原酶/分散酶和胰蛋白酶均增强了病毒感染，表明蛋白酶预处理可能是增强许多体内应用中病毒介导的基因转导的有用策略。
