CAR T Cells Secreting NGF-Neutralizing scFv Enhance Efficacy in Clear Cell Renal Cell Carcinoma by Relieving Immunosuppression through Immunosympathectomy [scRNA-seq]. CAR T Cells Secreting NGF-Neutralizing scFv Enhance Efficacy in Clear Cell Renal Cell Carcinoma by Relieving Immunosuppression through Immunosympathectomy [scRNA-seq]

This study investigated the role of sympathetic denervation in enhancing CAR T cell anti-tumor efficacy in renal clear cell carcinoma. By blocking the NGF pathway with antibodies, we demonstrated reduced sympathetic nerve distribution and delayed tumor progression. Furthermore, CAR T cells engineered to secrete NGF scFv achieved tumor immunosympathectomy, leading to improved anti-tumor effects. RNA sequencing revealed that this enhancement was linked to reduced terminal exhaustion in CD8 T cells and inhibited macrophage polarization from M1 to M2, promoting a robust anti-tumor immune state. Additionally, splenic T cells exhibited a stronger immune effector phenotype following the infusion of NGF scFv-secreting CAR T cells. These findings suggest that immunosympathectomy may represent a novel strategy to mitigate tumor-induced immunosuppression and improve CAR T cell efficacy in solid tumors. Overall design: Tumor cells were inoculated into the left axilla of Balb/c mice. When tumors reached approximately 80 mm3, mice were intraperitoneally injected with 200 mg/kg cyclophosphamide. Two days later, the mice were randomly divided into three groups and intravenously infused with either UT, 1×107 V28z, or 1×107 V28z/αNGF cells. 500,000 U of human recombinant IL-2 was administered intraperitoneally, followed by continuous injections twice daily for two days.

本研究探讨了交感神经去支配在肾透明细胞癌模型中增强CAR-T细胞抗肿瘤功效的作用机制。通过抗体阻断神经生长因子（NGF, Nerve Growth Factor）通路，我们证实可减少交感神经分布并延缓肿瘤进展。进一步研究发现，经工程化改造以分泌NGF单链可变片段（scFv, single-chain variable fragment）的CAR-T细胞能够实现肿瘤免疫交感神经切除，进而显著提升抗肿瘤效应。RNA测序结果显示，该疗效增强与CD8阳性T细胞的终末耗竭减少、巨噬细胞从M1型向M2型极化受抑制密切相关，可促成强效的抗肿瘤免疫状态。此外，输注分泌NGF scFv的CAR-T细胞后，小鼠脾脏T细胞呈现出更强的免疫效应细胞表型。本研究结果表明，免疫交感神经切除或可成为缓解肿瘤诱导的免疫抑制、提升实体瘤中CAR-T细胞治疗疗效的全新策略。实验整体设计如下：将肿瘤细胞接种于BALB/c小鼠左侧腋窝。当肿瘤体积约达80 mm³时，对小鼠腹腔注射200 mg/kg环磷酰胺。两天后，将小鼠随机分为三组，分别静脉输注UT、1×10⁷个V28z细胞或1×10⁷个V28z/αNGF细胞。随后腹腔给予500,000单位人重组IL-2，之后每日连续两次注射，持续两日。