PO.CL05.04 · 临床研究
超越密度：拓扑感知的空间指标预测免疫治疗结局
Beyond density: Topography-aware spatial metrics predict immunotherapy outcomes
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：越来越多的证据表明，肿瘤-免疫生态系统的空间组织决定了检查点抑制剂的获益。因此，对基线细胞丰度进行校正的拓扑感知指标更适合在多个尺度上量化空间免疫模式，如吸引、排斥和聚集。我们假设此类多尺度空间指标在解释晚期非小细胞肺癌（NSCLC）患者的治疗获益和生存方面优于单纯的免疫细胞密度。
方法：我们对54例接受检查点抑制剂治疗的NSCLC患者的诊断组织应用了多重免疫荧光流程。标注的细胞类别包括癌细胞、CD8⁺、CD8⁺FoxP3⁺、M1和M2巨噬细胞（-/+PDL1）、CD68-CD163+细胞以及基质非免疫细胞。为在空间上解析免疫模式，我们计算了邻域富集（ENR）和径向局部富集（RLE）。ENR在k近邻域（k=5-100）上计算，提供了两种细胞类别之间的吸引倾向信息，且独立于细胞密度。RLE在10-50 μm半径上评估，反映空间细胞聚集倾向。空间指标与治疗反应和总生存相关联。
结果：
CD8⁺FoxP3⁺细胞对风险进行分层。应答者表现出CD8⁺或CD8⁺PD1⁺细胞在不同尺度上与CD8⁺FoxP3⁺相邻（ENR k=5-100，p=0.04-0.007），在较短距离处最强。在生存分析中，CD8⁺FoxP3⁺细胞越接近PDL1+癌细胞时与更短的生存相关（ENR：HR 3.7-7.5，p=0.01-0.04）。
基质中CD8的几何分布追踪反应但不追踪生存。CD8⁺PD1⁺聚集程度的增加与更高的治疗反应相关（ENR k=10-100，p=0.05-0.002）。值得注意的是，CD8⁺PD1-细胞未显示此类关联。这种对治疗反应的积极影响在本队列中并未转化为更长的总生存。
肿瘤纯度和致密度与更短的生存相关。癌细胞在10-50 μm内的局部富集与不良反应和更短的OS相关（RLE：HR 2.2-2.5，p=0.03-0.007），提示更致密、均质的肿瘤巢是一个风险因素。
结论：拓扑感知的空间指标捕捉到了超越简单密度的具有临床意义的组织结构。这些发现支持对生物标志物进行依赖细胞背景的评估。此外，我们的发现还为聚焦评估一种罕见的、迄今描述较少的CD8⁺FoxP3⁺免疫细胞亚型提供了原位证据。
查看英文原文 English abstract
Background: Accumulating evidence suggests that the spatial organization of tumor-immune ecosystems determines the benefit of checkpoint inhibitors. Therefore, topography-aware metrics that adjust for baseline cell abundance are better suited to quantify spatial immune patterns, such as attraction, repulsion, and clustering, across scales. We hypothesized that such multiscale spatial metrics would outperform immune cell density alone for explaining therapy benefit and survival in advanced non-small cell lung cancer (NSCLC) patients.
Methods: We applied a multiplex immunofluorescence pipeline on diagnostic tissue of 54 NSCLC patients treated with checkpoint inhibitors. Annotated cell classes included cancer cells, CD8⁺, CD8⁺FoxP3⁺, M1 and M2 macrophages (-/+PDL1), CD68 - CD163 + cells and stromal non-immune cells. To spatially resolve the immune patterns, we computed neighborhood enrichment (ENR) and radial local enrichment (RLE). ENR was computed over k-nearest neighborhoods (k=5-100) and provided information about the attraction tendency between two cell classes, independent from cell density. RLE was evaluated over 10-50 µm radii and reflected the spatial cell clustering tendency. The spatial metrics were associated with therapy response and overall survival.
Results:
CD8 ⁺ FoxP3 ⁺ cells stratify risk. Responders demonstrate neighboring of CD8⁺ or CD8⁺PD1⁺ cells to CD8⁺FoxP3⁺ at different scales (ENR k=5-100, p=0.04-0.007), strongest at shorter distances. In the survival analysis, CD8⁺FoxP3⁺ cells were associated with shorter survival when they were closer to PDL1+ cancer cells (ENR: HR 3.7-7.5, p=0.01-0.04).
CD8 geometry in stroma tracks response but not survival. Increased clustering of CD8⁺PD1⁺ associated with higher response to therapy (ENR k=10-100, p=0.05- 0.002). Notably, CD8⁺PD1 - cells did not show such associations. This positive impact on therapy response did not translate into a longer overall survival in this cohort.
Tumor purity and compactness is associated with shorter survival. Local enrichment of cancer cells within 10-50 µm was associated with poor response and shorter OS, (RLE: HR 2.2-2.5, p=0.03-0.007), pointing to denser, homogeneous tumor nests as a risk factor.
Conclusion: Topography-aware spatial metrics capture clinically meaningful organization beyond simple density. These findings supported the cellular context-dependent evaluation of biomarkers. Furthermore our findings also provide in situ evidence for a focused evaluation of a rare, hitherto poorly described subtype of CD8⁺FoxP3⁺ immune cells.
利益披露 Disclosure
A. Mezheyeuski, None..
N. Hekmati, None..
I. Hrynchyk, None..
A. Lindberg, None..
J. Friedrich, None..
J. Mattsson, None..
M. Gulyas, None..
K. Kärre, None..
J. Isaksson, None..
C. Strell, None..
P. Micke, None.