Oncolytic viruses (OVs) offer a novel approach to treat solid tumors; however, their efficacy is frequently suboptimal due to various limiting factors. To address this challenge, we engineered an OV containing targets for neuron-specific microRNA-124 and Granulocyte-macrophage colony-stimulating factor (GM-CSF), significantly enhancing its neuronal safety while minimally compromising its replication capacity. Moreover, we identified PARP1 as an HSV-1 replication restriction factor using genome-wide CRISPR screening. In models of glioblastoma (GBM) and triple-negative breast cancer (TNBC), we showed that the combination of OV and a PARP inhibitor (PARPi) exhibited superior efficacy compared to either monotherapy. Additionally, single-cell RNA sequencing (scRNA-seq) revealed that this combination therapy sensitized TNBC to immune checkpoint blockade, and the incorporation of an immune checkpoint inhibitor (ICI) further increased the survival rate of tumor-bearing mice. The combination of PARPi and ICI synergistically enhanced the ability of OV to establish durable tumor-specific immune responses. Our study effectively overcomes the inherent limitations of OV therapy, providing valuable insights for the clinical treatment of TNBC, GBM, and other malignancies.

中文翻译：

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使用全基因组 CRISPR 筛选鉴定参与溶瘤病毒疗法的限制性分子


溶瘤病毒 （OVs） 提供了一种治疗实体瘤的新方法;然而，由于各种限制因素，它们的疗效往往不理想。为了应对这一挑战，我们设计了一种包含神经元特异性 microRNA-124 和粒细胞-巨噬细胞集落刺激因子 （GM-CSF） 靶标的 OV，显著提高了其神经元安全性，同时对其复制能力的影响最小。此外，我们使用全基因组 CRISPR 筛选将 PARP1 确定为 HSV-1 复制限制因子。在胶质母细胞瘤 （GBM） 和三阴性乳腺癌 （TNBC） 的模型中，我们表明 OV 和 PARP 抑制剂 （PARPi） 的组合与任何一种单一疗法相比表现出更好的疗效。此外，单细胞 RNA 测序 （scRNA-seq） 显示，这种联合疗法使 TNBC 对免疫检查点阻断敏感，免疫检查点抑制剂 （ICI） 的加入进一步提高了荷瘤小鼠的存活率。PARPi 和 ICI 的组合协同增强了 OV 建立持久的肿瘤特异性免疫反应的能力。我们的研究有效克服了 OV 疗法的固有局限性，为 TNBC、GBM 和其他恶性肿瘤的临床治疗提供了有价值的见解。