治疗耐药性是免疫检查点抑制剂实现长期获益的主要障碍。新辅助抗PD-1耐药的潜在机制仍不清楚。使用队列中切除的肿瘤样本进行多组学分析，包括质谱流式细胞术、单细胞RNA-seq、bulk RNA-seq和多色流式细胞术的非小细胞肺癌 (NSCLC) 患者接受了新辅助抗 PD-1 治疗。从未经治疗的患者获得的肿瘤和配对肺部样本用作对照。使用从新鲜组织和肺癌细胞系中分离的原代细胞进行体外实验。体内实验采用Lewis小鼠模型。组织驻留记忆CD8 T细胞（CD8 TRM）的数量、分化状态和克隆扩增与新辅助抗PD-1治疗的疗效呈正相关。人类非小细胞肺癌。相反，未成熟CD1c经典2型树突状细胞（imcDC2）和galectin-9癌细胞的数量与治疗效果呈负相关。发现了一个上皮/imDC2 抑制轴通过 galectin-9/TIM-3 抑制 CD8 TRM 的抗肿瘤反应。 CD8 TRMs 和半乳糖凝集素 9 癌细胞相关基因的表达水平可预测人类 NSCLC 患者抗 PD-1 新辅助治疗的临床结果。最后，阻断TIM-3和PD-1可以通过促进imcDC2和CD8 TRM介导的肿瘤杀伤的抗原呈递来提高荷瘤小鼠的存活率。表达Galectin-9的肿瘤细胞维持新辅助抗肿瘤药物的原发耐药性。通过半乳糖凝集素 9/TIM-3 介导的 imcDC2 和 CD8 TRM 抑制来治疗 NSCLC。补充抗 TIM-3 可以打破上皮/imcDC2/CD8 TRM 抑制环，从而克服抗 PD-1 耐药性。NCT03732664.© 作者（或其雇主）2024。CC 允许重复使用BY-NC。禁止商业再利用。请参阅权利和权限。英国医学杂志出版。

Therapeutic resistance is a main obstacle to achieve long-term benefits from immune checkpoint inhibitors. The underlying mechanism of neoadjuvant anti-PD-1 resistance remains unclear.Multi-omics analysis, including mass cytometry, single-cell RNA-seq, bulk RNA-seq, and polychromatic flow cytometry, was conducted using the resected tumor samples in a cohort of non-small cell lung cancer (NSCLC) patients received neoadjuvant anti-PD-1 therapy. Tumor and paired lung samples acquired from treatment-naïve patients were used as a control. In vitro experiments were conducted using primary cells isolated from fresh tissues and lung cancer cell lines. A Lewis-bearing mouse model was used in the in vivo experiment.The quantity, differentiation status, and clonal expansion of tissue-resident memory CD8+ T cells (CD8+ TRMs) are positively correlated with therapeutic efficacy of neoadjuvant anti-PD-1 therapy in human NSCLC. In contrast, the quantity of immature CD1c+ classical type 2 dendritic cells (imcDC2) and galectin-9+ cancer cells is negatively correlated with therapeutic efficacy. An epithelium/imDC2 suppressive axis that restrains the antitumor response of CD8+ TRMs via galectin-9/TIM-3 was uncovered. The expression level of CD8+ TRMs and galectin-9+ cancer cell-related genes predict the clinical outcome of anti-PD-1 neoadjuvant therapy in human NSCLC patients. Finally, blockade of TIM-3 and PD-1 could improve the survival of tumor-bearing mouse by promoting the antigen presentation of imcDC2 and CD8+ TRMs-mediated tumor-killing.Galectin-9 expressing tumor cells sustained the primary resistance of neoadjuvant anti-PD-1 therapy in NSCLC through galectin-9/TIM-3-mediated suppression of imcDC2 and CD8+ TRMs. Supplement of anti-TIM-3 could break the epithelium/imcDC2/CD8+ TRMs suppressive loop to overcome anti-PD-1 resistance.NCT03732664.© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.