Department of Immunology, Genetics, and Pathology, Science for Life Laboratory, Uppsala University, Uppsala SE-75185, Sweden.
Hum Gene Ther. 2013 Aug;24(8):766-75. doi: 10.1089/hum.2012.132.
Secretogranin III (SGC3) belongs to the granin family and is highly expressed in endocrine and neural tissues. The human SCG3 promoter has not yet been characterized. We identified that a 0.5-kb DNA fragment upstream of the SCG3 gene can selectively drive transgene expression in neuroblastoma cell lines. The strength of transgene expression was further increased, with specificity maintained, by addition of the human achaete-scute complex homolog 1 (ASH1) enhancer. We developed an oncolytic serotype 5-based adenovirus, in which the SCG3 promoter and ASH1 enhancer drive E1A gene expression. The virus was further modified with a cell-penetrating peptide (Tat-PTD) in the viral capsid, which we have previously shown results in increased adenovirus transduction efficiency of many neuroblastoma cell lines. The virus, Ad5PTD(ASH1-SCG3-E1A), shows selective and efficient killing of neuroblastoma cell lines in vitro, including cisplatin-, etoposide-, and doxorubicin-insensitive neuroblastoma cells. Furthermore, it delays tumor growth and thereby prolonged survival for nude mice harboring subcutaneous human neuroblastoma xenograft. In conclusion, we report a novel oncolytic adenovirus with potential use for neuroblastoma therapy.
分泌颗粒蛋白 III（SGC3）属于颗粒蛋白家族，在内分泌和神经组织中高度表达。人类 SCG3 启动子尚未被表征。我们发现，SCG3 基因上游的 0.5kb DNA 片段可以选择性地驱动神经母细胞瘤细胞系中转基因的表达。通过添加人 Achaete-scute 复合物同源物 1（ASH1）增强子，进一步增加了转基因的表达强度，同时保持了特异性。我们开发了一种基于溶瘤 5 型腺病毒的病毒，其中 SCG3 启动子和 ASH1 增强子驱动 E1A 基因的表达。该病毒进一步在病毒衣壳中修饰了一个穿透肽（Tat-PTD），我们之前已经证明，这会增加许多神经母细胞瘤细胞系中腺病毒的转导效率。该病毒，Ad5PTD（ASH1-SCG3-E1A），在体外对神经母细胞瘤细胞系具有选择性和高效的杀伤作用，包括对顺铂、依托泊苷和阿霉素不敏感的神经母细胞瘤细胞。此外，它还延迟了肿瘤的生长，从而延长了携带皮下人神经母细胞瘤异种移植物的裸鼠的存活时间。总之，我们报告了一种新型的溶瘤腺病毒，具有用于神经母细胞瘤治疗的潜力。
