Glioblastoma (GBM) is an immunosuppressive, highly vascular and devastating malignant brain tumor. Even with progressive combination treatment that includes surgery, radiotherapy, and chemotherapy, the prognosis for GBM patients is still extremely poor. Oncolytic adenovirus (OAd) can specifically replicate in GBM cells, permitting the rapid copy of the therapeutic genes it carries. Moreover, E1A is an essential gene in adenoviral replication and is the first gene expressed upon viral infection. E1A expression can be regulated by the Ki67 promoter, while the CMV promoter drives therapeutic gene expression. However, the efficacy of a double-controlled OAd driven by the Ki67 core promoter and armed with IL-15 against GBM cells has not been investigated. Fluorescence microscopy was performed to evaluate infection ability in the viruses. Cell viability was detected by CCK-8 assay. Levels of cytokines in different supernatants were determined by ELISA, and IL-15 gene expression was measured by RT-PCR. Angiogenic capacity was analyzed by tube formation assay. We successfully constructed a double-controlled oncolytic adenovirus driven by the Ki67 core promoter and armed with IL-15 that selectively infected and killed GBM cells while sparing normal cells. The adenoviruses prime IL-15 gene expression to significantly enhance anti-GBM efficacy through effective activation of microglial cells. Moreover, OAd not only directly inhibits angiogenesis but exhibits potent antiangiogenic capacity mediated by the reduction of VEGF secretion. These results provide new insight into the effects of a novel double-controlled OAd driven by the Ki67 core promoter and armed with IL-15 in glioblastoma treatment, which may help in the development of novel therapies in solid tumors.

中文翻译：

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由 Ki67 核心启动子驱动并配备 IL-15 的新型双控溶瘤腺病毒对胶质母细胞瘤细胞的疗效

胶质母细胞瘤 (GBM) 是一种免疫抑制性、高度血管性和破坏性的恶性脑肿瘤。即使采用包括手术、放疗和化疗在内的渐进性联合治疗，GBM患者的预后仍然极差。溶瘤腺病毒 (OAd) 可以在 GBM 细胞中特异性复制，从而允许快速复制其携带的治疗基因。此外，E1A 是腺病毒复制的必需基因，是病毒感染后第一个表达的基因。E1A 表达可由 Ki67 启动子调节，而 CMV 启动子驱动治疗性基因表达。然而，尚未研究由 Ki67 核心启动子驱动并配备 IL-15 的双重控制 OAd 对 GBM 细胞的功效。进行荧光显微镜检查以评估病毒的感染能力。通过CCK-8测定检测细胞活力。ELISA测定不同上清液中细胞因子水平，RT-PCR测定IL-15基因表达。通过管形成测定分析血管生成能力。我们成功构建了由 Ki67 核心启动子驱动并配备 IL-15 的双重控制溶瘤腺病毒，该病毒选择性感染和杀死 GBM 细胞，同时保留正常细胞。腺病毒引发 IL-15 基因表达，通过有效激活小胶质细胞显着增强抗 GBM 功效。此外，OAd 不仅直接抑制血管生成，而且表现出由 VEGF 分泌减少介导的有效抗血管生成能力。