PO.BCS01.14 · 生物信息与计算
靶向主调控因子以重编程中性粒细胞并增强去势抵抗性前列腺癌中 PD-1 阻断的疗效
Targeting master regulators to reprogram neutrophils and enhance PD-1 blockade efficacy in castration-resistant prostate cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
引言：中性粒细胞介导的免疫抑制限制了去势抵抗性前列腺癌（CRPC）的免疫治疗，但其机制尚不明确。本研究旨在识别这种免疫抑制的驱动因素，并评估靶向主调控因子（MR）蛋白是否能够重编程中性粒细胞、重塑肿瘤微环境（TME）并增强 PD-1 阻断的疗效。
方法：我们使用经 MR 抑制剂和抗 PD-1 疗法处理的原位 CRPC 模型，评估中性粒细胞耗竭和免疫重塑。借助 5,000 重 Xenium 面板进行空间图谱分析，并通过精心整理的 scRNA-seq 图谱进行细胞类型注释，揭示了组织组织结构的详细图谱。对免疫细胞群及空间重组（尤其是中性粒细胞）的治疗效果进行了量化。基于 VIPER 的分析定义了 MR 驱动的中性粒细胞功能状态，并使用 Xenium 数据进行了空间映射。最后，通过细胞间通讯网络分析探索了中性粒细胞与肿瘤细胞之间的配体-受体相互作用。
未发表数据摘要：我们在四个治疗组（载体对照、Trametinib、抗 PD-1 和联合治疗）中开展了一项 Xenium 试点研究，通过细胞丰度和空间重组来评估 MR 抑制对免疫反应的影响。抗 PD-1 诱导了广泛的免疫细胞募集，尤其是中性粒细胞，而 Trametinib 的影响甚微。联合治疗将中性粒细胞丰度降至基线以下，减弱了检查点阻断诱导的中性粒细胞聚集。NK/T 细胞、单核细胞和树突状细胞增多，而巨噬细胞保持稳定。空间分析显示，抗 PD-1 导致肿瘤细胞边缘化，而联合治疗则部分恢复了整合。中性粒细胞从边缘化转变为在肿瘤细胞周围形成免疫屏障，而 NK/T 细胞主要与基质/髓系细胞相互作用，表明存在免疫排斥。尽管所有治疗均上调了 PD-L1，但联合治疗诱导了 PD-L1 表达而未形成屏障，提示是空间组织结构而非仅仅是配体表达，驱动了中性粒细胞介导的免疫排斥。
结论：这些治疗在不破坏组织结构的情况下重塑了肿瘤-免疫构架，依赖于免疫细胞丰度及其在 TME 内的空间分布。抗 PD-1 诱导广泛的免疫浸润，但中性粒细胞形成限制肿瘤穿透的屏障，而 MR 抑制剂 Trametinib 可缓解这一屏障。中性粒细胞在所有治疗中均上调 PD-L1，并在抗 PD-1 治疗后达到峰值，而 Trametinib 和联合治疗则驱动 PD-L1 表达而不形成屏障。
查看英文原文 English abstract
Introduction: Neutrophil-mediated immunosuppression limits immunotherapy in Castration-Resistant Prostate Cancer (CRPC), but its mechanisms are unclear. This study aims to identify the drivers of this immunosuppression and evaluate whether targeting Master Regulator (MR) proteins can reprogram neutrophils, remodel the tumor microenvironment (TME), and enhance PD-1 blockade efficacy.
Methods: We used orthotopic CRPC models treated with MR inhibitors and anti-PD-1 therapy to assess neutrophil depletion and immune remodeling. Spatial profiling with the 5,000-plex Xenium panel along with cell types annotation via a curated scRNA-seq atlas, revealed a detailed map of tissue organization. Treatment effects on immune populations and spatial reorganization, particularly neutrophils, were quantified. VIPER-based analysis defined MR-driven neutrophil functional states, and spatial mapping was conducted using Xenium data. Finally, ligand-receptor interactions between neutrophils and tumor cells were explored through cell-cell communication network analysis.
Summary of unpublished data: We conducted a pilot Xenium study across four treatment arms (vehicle, Trametinib, anti-PD1, and combination) to assess MR inhibition's effect on immune responses through cell abundance and spatial reorganization. Anti-PD1 induced broad immune recruitment, especially neutrophils, while Trametinib had minimal effect. Combination therapy reduced neutrophil abundance below baseline, attenuating checkpoint blockade-induced neutrophil accumulation. NK/T cells, monocytes, and dendritic cells increased, while macrophages remained stable. Spatial analysis showed that anti-PD1 caused tumor cells to marginalize, while combination therapy partially restored integration. Neutrophils transitioned from marginalization to forming an immunological barrier around tumor cells, and NK/T cells interacted mainly with stromal/myeloid cells, indicating immune exclusion. Despite upregulating PD-L1 in all treatments, combination therapy induced PD-L1 expression without barrier formation, suggesting that spatial organization, not just ligand expression, drives neutrophil-mediated immune exclusion.
Conclusion: Treatments reshape tumor-immune architecture without disrupting tissue organization, relying on both immune cell abundance and their spatial distribution within the TME. Anti-PD1 induces broad immune infiltration, but neutrophils form a barrier limiting tumor penetration, which is relieved by the MR-inhibitor Trametinib. Neutrophils upregulate PD-L1 across all treatments, peaking after anti-PD1, while Trametinib and combination therapy drive PD-L1 expression without barrier formation.
利益披露 Disclosure
M. Franchini, None..
F. Picech, None..
C. Abate-Shen, None.
A. Califano,
DarwinHealth Independent Contractor, Stock.