Integrative Analysis of Targeted Genomic Profiling and Immune Cell Infiltration in the Prognosis of Lung Adenocarcinoma

The prognosis of lung adenocarcinoma (LUAD) is traditionally evaluated via tumor-node-metastasis (TNM) staging, which does not account for the patient's immune status. This protocol integrates genomic profiling and immune microenvironment analysis to facilitate a more comprehensive postoperative prognostic evaluation. The method involves a retrospective analysis of paraffin-embedded tumor tissues using two primary techniques. First, next-generation sequencing (NGS) is performed with a customized 37-gene panel to identify mutations in driver genes and variants of uncertain significance. Second, multiplex immunofluorescence (mIF) is utilized to target markers including HLA-DR, CD68, CD163, CD206, PD-L1, and PanCK. This enables the quantification of spatial distribution and density for specific immune cell subpopulations across various tumor regions. This integrated approach enables the simultaneous assessment of genomic heterogeneity and tumor-infiltrating immune cell characteristics. The resulting data identifies specific combinations of mutational profiles-such as EGFR status-and immune cell densities. These integrated combinations enable the study of their collective impact on patient survival, offering a promising approach for the development of future lung cancer prognostic models. This protocol demonstrates a robust method for characterizing the complex biological features of the LUAD tumor microenvironment.