Figure 6 from Spatial Architecture of Myeloid and T Cells Orchestrates Immune Evasion and Clinical Outcome in Lung Cancer

Neutrophil infiltration is associated with recent subclonal expansion and poorer disease-free survival. A, Comparison of neutrophil cell density in primary tumors with metastasis-seeding clones detectable at time of surgery or during follow-up (metastasizing tumors) to tumors from patients who were metastasis-free and recurrence-free for more than 3 years of follow-up time (control) in LUAD (n = 28) and LUSC (n = 15) within the TRACERx discovery cohort. Maximum neutrophil density taken for tumors with multiple tumor cores. P values derived from one-tailed Wilcoxon test. B, Kaplan–Meier curves for DFS according to tumor-level TAN score in the validation cohort (n = 332 patients). P value derived from univariate Cox model adjusted for histology. C, Multivariable Cox proportional hazard regression analysis of DFS using tumor-level TAN score and tumor-level necrosis evaluation from H&E images of diagnostic tumor blocks. D, Recent subclonal expansion score, measured as the maximum cancer cell fraction of subclones at the terminus of the phylogenetic tree, compared between TAN-high and TAN-low tumor regions in LUAD and LUSC patients from the TRACERx 421 cohort. P values were derived from a linear mixed effects model with patient as random effect. E, Spatial ITH score of cell types and communities and ITH probabilities of TME classes in multiregion analysis (pan-immune n = 41 tumors, 112 cores). ITH score was calculated as the average standard deviation of the cell/community density in multiple regions per tumor and z-score transformed. ITH score of αSMA⁺ cells was derived from T cells and stroma panel (n = 39 tumors, 105 cores). ITH probability was calculated as 1 − probability of all regions having the same indicated TME class. F, Example phylogenetic tree depicting a stage IIIA LUSC case with a clonal PIK3CA mutation, high TAN scores, and recent subclonal expansion, including in a region (R5) that seeded a lymph node (LN) metastasis (FLN, FFPE LN). The metastasis-seeding lineage is highlighted in orange. Tumor-level TAN scores and regional subclonal expansion (SubExp) scores are reported for primary tumor regions. The reported TAN score represents the maximum of the tumor nest and stroma scores. Each cluster in the phylogenetic tree is assigned a color that is also represented in the region clone maps. The clone maps illustrate the prevalence of each clone within a region. G, IMC images shown for R5 and the LN metastasis. Scale bar, 200 μm. H, Summary schematic of the link between tumors with a neutrophil enriched microenvironment with tumor progression. ·, P < 0.1; *, P < 0.05; **, P < 0.01; ***, P < 0.001; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; ITH, intratumor heterogeneity.