胶质母细胞瘤（GBM）是一种高度侵袭性的脑癌，存活率低，促使人们探索新的治疗策略。免疫检查点抑制剂在癌症治疗中显示出前景，但与免疫相关的毒性和大脑渗透有关。在这里，我们提出了一种靶向方法，使用腺相关病毒血清型 9 (AAV9) 将抗 PD-1 的单链片段可变抗体 (scFv-PD-1) 系统性递送到肿瘤微环境 (TME) 中。单细胞 RNA 测序分析显示 GBM TME 中 PD-1 表达强劲，主要在 T 细胞上。 AAV9-scFv-PD-1表达并分泌scFv-PD-1，其有效结合PD-1。在免疫活性 GBM 小鼠模型中全身施用 AAV9-scFv-PD-1 会导致肿瘤部位的溶细胞 T 细胞强烈激活，其标志是 IFN-γ 和颗粒酶 B 的积累，从而导致肿瘤生长显着减少。重要的是，AAV9-scFv-PD-1 治疗可带来生存获益，凸显其治疗潜力。这项研究证明了系统性递送 AAV9 介导的 scFv-PD-1 局部表达用于 GBM 靶向免疫治疗的可行性，并值得进一步研究进行临床转化。© 2024。作者获得 Springer Science Business Media 的独家许可， LLC，施普林格自然集团的一部分。

Glioblastoma (GBM) is a highly aggressive brain cancer with a low survival rate, prompting the exploration of novel therapeutic strategies. Immune checkpoint inhibitors have shown promise in cancer treatment but are associated with immune-related toxicities and brain penetration. Here, we present a targeted approach using an adeno-associated virus serotype 9 (AAV9) to systemically deliver a single-chain fragment variable antibody against PD-1 (scFv-PD-1) into the tumor microenvironment (TME). Single-cell RNA sequencing analysis revealed robust PD-1 expression in GBM TME, predominantly on T cells. AAV9-scFv-PD-1 expressed and secreted scFv-PD-1, which effectively binds to PD-1. Systemic administration of AAV9-scFv-PD-1 in an immunocompetent GBM mouse model resulted in a robust cytolytic T-cell activation at the tumor site, marked by accumulation of IFN-γ and Granzyme B, leading to a significant reduction in tumor growth. Importantly, AAV9-scFv-PD-1 treatment conferred a survival benefit, highlighting its therapeutic potential. This study demonstrates the feasibility of systemically delivered AAV9-mediated local expression of scFv-PD-1 for targeted immunotherapy in GBM and warrants further investigation for clinical translation.© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.