Spatial proteomic profiling of tumor and stromal compartments in non-small-cell lung cancer identifies signatures associated with overall survival.

Objectives: Non-small-cell lung carcinoma (NSCLC) is the most prevalent and lethal form of lung cancer. The need for biomarker-informed stratification of targeted therapies has underpinned the need to uncover the underlying properties of the tumor microenvironment (TME) through high-plex quantitative assays.

Methods: In this study, we profiled resected NSCLC tissues from 102 patients by targeted spatial proteomics of 78 proteins across tumor, immune activation, immune cell typing, immune-oncology, drug targets, cell death and PI3K/AKT modules to identify the tumor and stromal signatures associated with overall survival (OS).

Results: Survival analysis revealed that stromal CD56 (HR = 0.384,  = 0.06) and tumoral TIM3 (HR = 0.703,  = 0.05) were associated with better survival in univariate Cox models. In contrast, after adjusting for stage, BCLXL (HR = 2.093,  = 0.02) and cleaved caspase 9 (HR = 1.575,  = 0.1) negatively influenced survival. Delta testing indicated the protective effect of TIM-3 (HR = 0.614,  = 0.04) on OS. In multivariate analysis, CD56 (HR = 0.172,  = 0.001) was associated with better survival in the stroma, while B7.H3 (HR = 1.72,  = 0.008) was linked to poorer survival in the tumor.

Conclusions: Deciphering the TME using high-plex spatially resolved methods is giving us new insights into compartmentalised tumor and stromal protein signatures associated with clinical endpoints in NSCLC.