Honigsberg Rachel, Cruz Tatiana, Yoffe Liron, Tang Meixian Stephanie, Dicle Ozge, Markowitz Geoffrey, Michael Marissa, Singh Arshdeep, Altorki Nasser K, Elemento Olivier, Villena-Vargas Jonathan
Meining School of Biomedical Engineering, Cornell University, Ithaca, NY, USA.
Department of Cardiothoracic Surgery, Weill Cornell Medicine, New York, NY, USA.
bioRxiv. 2025 Jun 1:2025.04.27.650862. doi: 10.1101/2025.04.27.650862.
Elucidating the anti-tumor role of tumor-draining lymph nodes (tdLNs) in patients could offer critical mechanistic insight and shift therapeutic strategies from a tumor-centric approach to one that considers tumor-immune system interplay. Our study characterizes benign tdLNs T cell anti-tumor responses beyond initial T cell priming in patients with resectable non-small cell lung cancer. We further investigated whether tumor-specific tdLN T cells were altered by immune checkpoint blockade (ICB) locally and systemically. We performed single-cell TCR lineage tracing and transcriptomic profiling on 672,886 CD8 T cells from 41 tumor, benign tdLN, and blood samples in 14 patients treated with or without neoadjuvant chemoimmunotherapy (ChemoIO). Using deep-integrating clonal tracking with machine learning-based transcriptional analysis, our findings revealed that benign tdLNs locally and independently orchestrate two transcriptionally distinct tumor-specific memory CD8 T cell populations: one with ZNF683+ CXCR6+ tumor tissue-residency potential, and another with cytotoxic memory potential. Furthermore, tdLN-derived clones not only constitute the dominant tumor-infiltrating (75%) and circulating (>90%) tumor-specific expanded T cell populations but also preserve their transcriptionally distinct subset identities within the tumor T cell effector state. ChemoIO selectively increased the clonal diversity and cytotoxic memory/TEMRA programs of tdLN-derived clones locally and systemically, both of which remained unchanged in clones lacking tdLN TCR lineage. In conclusion, the tdLN locally orchestrates tumor-reactive and ChemoIO-reactive transcriptional distinct T cell subsets that shape the circulating blood and tumor T cell environments. These findings represent a clinical paradigm shift with implications regarding the extent of tdLN resection during surgery, timing of ChemoIO treatment, and the development of memory T cell-based immunotherapies.
阐明肿瘤引流淋巴结（tdLNs）在患者体内的抗肿瘤作用，可为关键机制提供见解，并将治疗策略从以肿瘤为中心的方法转变为考虑肿瘤-免疫系统相互作用的方法。我们的研究对可切除非小细胞肺癌患者的良性tdLNs T细胞抗肿瘤反应进行了表征，超越了初始T细胞致敏阶段。我们进一步研究了肿瘤特异性tdLN T细胞是否会因免疫检查点阻断（ICB）而在局部和全身发生改变。我们对14例接受或未接受新辅助化疗免疫治疗（ChemoIO）的患者的41个肿瘤、良性tdLN和血液样本中的672,886个CD8 T细胞进行了单细胞TCR谱系追踪和转录组分析。通过基于机器学习的转录分析进行深度整合克隆追踪，我们的研究结果表明，良性tdLNs在局部独立地协调两个转录上不同的肿瘤特异性记忆CD8 T细胞群体：一个具有ZNF683+ CXCR6+肿瘤组织驻留潜力，另一个具有细胞毒性记忆潜力。此外，tdLN衍生的克隆不仅构成了主要的肿瘤浸润（75%）和循环（>90%）肿瘤特异性扩增T细胞群体，而且在肿瘤T细胞效应状态下保持其转录上不同的亚群特征。ChemoIO在局部和全身选择性地增加了tdLN衍生克隆的克隆多样性和细胞毒性记忆/TEMRA程序，而在缺乏tdLN TCR谱系的克隆中这些程序保持不变。总之，tdLN在局部协调肿瘤反应性和ChemoIO反应性转录上不同的T细胞亚群，这些亚群塑造了循环血液和肿瘤T细胞环境。这些发现代表了一种临床范式转变，对手术期间tdLN切除的范围、ChemoIO治疗的时机以及基于记忆T细胞的免疫疗法的开发具有重要意义。
