Intracranial AAV-IFN-β gene therapy eliminates invasive xenograft glioblastoma and improves survival in orthotopic syngeneic murine model.
Intracranial AAV-IFN-β gene therapy eliminates invasive xenograft glioblastoma and improves survival in orthotopic syngeneic murine model.
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DOI：
10.1002/1878-0261.12020
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发表时间：
2017-02
影响因子：
6.6
通讯作者：
Sena-Esteves M
中科院分区：
文献类型：
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作者：
GuhaSarkar D;Neiswender J;Su Q;Gao G;Sena-Esteves M
胶质母细胞瘤（GBM）细胞的高度侵袭性和遗传异质性是目前标准治疗后肿瘤复发的主要原因，因此是死亡的直接原因。之前，我们已经证明，通过局部施用腺相关病毒载体（AAV）的颅内干扰素β（IFN-β）基因治疗成功治疗了非侵入性原位胶质母细胞瘤模型。在这里，我们通过在侵袭性人类GBM异种移植和同基因小鼠模型中测试这种方法来扩展这些发现。首先，我们证明了编码人IFN-β的AAV的单次颅内注射消除了侵袭性人GBM 8肿瘤并促进了长期存活。接下来，我们筛选了五种具有不同启动子的AAV-IFN-β载体，以在同基因GL 261肿瘤的背景下驱动小鼠IFN-β在脑中的安全表达。两种AAV-IFN-β载体由于安全性问题而被排除，但使用其他三种载体的治疗研究显示了广泛的肿瘤细胞死亡、肿瘤周围小胶质细胞的活化以及用AAV/P2-Int-mIFN-β载体治疗的动物的中位存活率增加56%。我们还评估了AAV-IFN-β疗法与替莫唑胺（TMZ）组合的治疗效果。由于TMZ影响DNA复制，这是一个对活性转录前单链AAV载体的第二链DNA合成至关重要的事件，我们测试了两种TMZ治疗方案。在AAV-IFN-β之前用TMZ治疗消除了后者的任何益处，而与对照相比，相反的治疗顺序使中位生存期增加了一倍。这些研究证明了颅内AAV-IFN-β治疗在高度迁移性GBM模型以及同系小鼠模型中的治疗潜力，并且与TMZ组合可能增强其抗肿瘤效力。
The highly invasive property of glioblastoma (GBM) cells and genetic heterogeneity are largely responsible for tumor recurrence after the current standard‐of‐care treatment and thus a direct cause of death. Previously, we have shown that intracranial interferon‐beta (IFN‐β) gene therapy by locally administered adeno‐associated viral vectors (AAV) successfully treats noninvasive orthotopic glioblastoma models. Here, we extend these findings by testing this approach in invasive human GBM xenograft and syngeneic mouse models. First, we show that a single intracranial injection of AAV encoding human IFN‐β eliminates invasive human GBM8 tumors and promotes long‐term survival. Next, we screened five AAV‐IFN‐β vectors with different promoters to drive safe expression of mouse IFN‐β in the brain in the context of syngeneic GL261 tumors. Two AAV‐IFN‐β vectors were excluded due to safety concerns, but therapeutic studies with the other three vectors showed extensive tumor cell death, activation of microglia surrounding the tumors, and a 56% increase in median survival of the animals treated with AAV/P2‐Int‐mIFN‐β vector. We also assessed the therapeutic effect of combining AAV‐IFN‐β therapy with temozolomide (TMZ). As TMZ affects DNA replication, an event that is crucial for second‐strand DNA synthesis of single‐stranded AAV vectors before active transcription, we tested two TMZ treatment regimens. Treatment with TMZ prior to AAV‐IFN‐β abrogated any benefit from the latter, while the reverse order of treatment doubled the median survival compared to controls. These studies demonstrate the therapeutic potential of intracranial AAV‐IFN‐β therapy in a highly migratory GBM model as well as in a syngeneic mouse model and that combination with TMZ is likely to enhance its antitumor potency.