Immune checkpoint blockade (ICB) has transformed the treatment of advanced non-small cell lung cancer (NSCLC), yet only a subset of patients derives durable benefit. Established biomarkers such as PD-L1 expression and tumor mutational burden have limited predictive accuracy and do not capture the temporal dynamics of the on-treatment immune response. We previously demonstrated that an early kinetic pattern of C-reactive protein (CRP), termed CRP-flare, is strongly associated with response and overall survival in patients with NSCLC receiving ICB. The biological basis of this clinical signal, however, remains incompletely understood. By integrating dynamic serum proteomics, multiplex cytokine profiling and single-cell transcriptomics of peripheral blood mononuclear cells (PBMC), this project aims to characterize the molecular and cellular processes underlying early treatment-associated immune responses and to develop a dynamic multi-omic classifier of ICB response.

Longitudinal cytokine and targeted serum proteomic analyses have already identified distinct early protein kinetics associated with radiological response, CRP-flare and survival. Building on these findings, deep serum proteomics will extend the analysis beyond predefined inflammatory markers and enable unbiased identification of dynamic protein signatures associated with treatment response. In parallel, longitudinal single-cell transcriptomics of PBMC will characterize changes in the circulating immune compartment and allow serum protein dynamics to be linked to specific immune-cell populations. Integration of these complementary molecular layers will be used to identify dynamic trajectories that reflect treatment response and to refine a multi-omic classifier.

The resulting classifier will be independently evaluated in a prospective NSCLC cohort with structured longitudinal collection of serum and PBMC samples. The overall goal of this project is the establishment of a dynamic serum- and immune-cell-based framework for response assessment in NSCLC, which could provide a minimally invasive and clinically applicable approach for early prediction and monitoring of benefit from ICB. By combining temporal molecular profiling with cellular characterization, the project aims to improve understanding of the biological basis of early treatment responses and to address the limitations of static biomarkers in clinical decision-making.