尽管免疫检查点抑制剂（ICI）的免疫疗法最近取得了进展，但许多非小细胞肺癌（NSCLC）患者在初始反应后未能做出反应或产生耐药性。使用工程病毒进行原位疫苗接种 (ISV) 已成为一种有前途的抗原不可知策略，它既可以调节肿瘤微环境 (TME)，又可以增强抗肿瘤 T 细胞反应以克服免疫抵抗。我们通过进行全基因组功能筛选并在流感基因组中引入八种干扰素 (IFN) 敏感突变，设计了一种减毒活病毒疫苗，即超干扰素敏感病毒 (HIS)。与野生型 (WT) 流感相比，HIS 复制在免疫功能正常的宿主中减弱，增强了其作为癌症治疗安全选择的潜力。 HIS ISV 在小鼠 NSCLC 中引发强烈但短暂的 I 型 IFN 反应，导致多功能效应 Th1 CD4 和细胞毒性 CD8 T 细胞富集到肿瘤中。在具有不同驱动突变和不同突变负荷的多种非小细胞肺癌同基因小鼠模型中，HIS ISV 与 WT 相比表现出增强的抗肿瘤功效。这种功效取决于宿主 1 型 IFN 反应和 T 淋巴细胞。 HIS ISV 克服了 LKB-1 缺陷型小鼠 NSCLC 中对抗 PD-1 的耐药性，从而提高了总体生存率并维持了全身肿瘤特异性免疫。这些研究提供了令人信服的证据，支持对 HIS 作为一种新型“现成”ISV 策略进行进一步临床评估，用于 ICI 难治性 NSCLC 患者。

Despite recent advances in immunotherapy with immune checkpoint inhibitors (ICI), many patients with non-small cell lung cancer (NSCLC) fail to respond or develop resistance after an initial response. In situ vaccination (ISV) with engineered viruses has emerged as a promising antigen-agnostic strategy that can both condition the tumor microenvironment (TME) and augment anti-tumor T cell responses to overcome immune resistance. We engineered a live attenuated viral vaccine, Hyper-Interferon Sensitive virus (HIS), by conducting a genome-wide functional screening and introducing eight interferon (IFN)-sensitive mutations in the influenza genome. Compared to wild-type (WT) influenza, HIS replication was attenuated in immunocompetent hosts, enhancing its potential as a safe option for cancer therapy. HIS ISV elicited robust yet transient type I IFN responses in murine NSCLCs, leading to an enrichment of polyfunctional effector Th1 CD4 and cytotoxic CD8 T cells into the tumor. HIS ISV demonstrated enhanced anti-tumor efficacy compared to WT in multiple syngeneic murine models of NSCLC with distinct driver mutations and varying mutational burden. This efficacy was dependent on host type 1 IFN responses and T lymphocytes. HIS ISV overcame resistance to anti-PD-1 in LKB-1 deficient murine NSCLC, resulting in improved overall survival and enduring systemic tumor-specific immunity. These studies provide compelling evidence to support further clinical evaluation of HIS as a novel 'off-the-shelf' ISV strategy for patients with NSCLC refractory to ICI.