Thanos Mourikis

Baseline gene expression with significant association with ICI treatment outcome (OS) in PD-L1 negative in both CANOPY-1 and Tempus cohort 1 patients

Criteria to define post-ICI patients (n = 326) into acquired (n = 75) and primary resistance (n = 59) in Tempus cohort

Table of differentially expressed gene table in acquired resistance compared to treatment naïve cohort

Distribution of biopsy tissue sites comparing the treatment-naïve group (cohort 1) with the post-ICI group (cohort 2) in the left two panels and contrasting primary resistance with acquired resistance within cohort 2 in the right two panels

List of gene expression signatured

Key patient characteristics of the five cohorts in the study

Analysis of TME between pre- and post-ICI samples in patients with paired biopsies (Tempus cohort 3).

Analysis of the TME after monochemotherapy in the Tempus cohort 3 and SU2C-NSCLC cohorts. A, Volcano plot of differential expression analysis for 12 patients treated with platinum-based chemotherapy within Tempus cohort 3. B, Box plot of IFNγ signature expression before and after platinum therapy in 12 patients with paired biopsies. C, Box plot of the IFNγ signature by prior platinum-based therapy history in the SU2C-NSCLC cohort. D, Schematic of PD-L1 IHC class switching observed in Tempus cohort 3 by treatment class (ICI vs. non-ICI).

Patient characteristics of three Tempus cohorts for age, gender, race and PD-L1 class

Immune checkpoint inhibitors (ICI) have demonstrated clinical efficacy in non–small cell lung cancer (NSCLC), and extensive research has been conducted to explore biomarkers predictive of ICI response. However, the impact of ICI on the tumor and tumor microenvironment at primary and acquired resistance states is understudied due to the difficulty of collecting tissue biopsies at disease progression. In this study, we leveraged clinical and real-world data to study ICI resistance. Data used in this work consist of treatment outcome information and tissue RNA sequencing data from advanced-stage NSCLC cohorts from three sources: the Tempus real-world evidence database; CANOPY-1 (NCT03631199), a phase III clinical trial in first-line NSCLC; and Stand Up To Cancer (SU2C) publication. Our results indicate higher IFNγ and T-cell exhaustion in patients’ tumors at acquired resistance and low levels of B-cell and dendritic cell expression at primary resistance. The lower B-cell and dendritic cell levels may be primarily driven by prior treatment with a platinum-based chemotherapy regimen. Baseline transcriptomics data additionally suggest that innate immune cells may play an antitumor role in PD-L1<1% patients, whereas IFNγ and T-cell inflammation are more predictive of ICI treatment outcomes in PD-L1≥1% patients. Our study suggests a clear divergence of the tumor microenvironment in patients with primary versus acquired resistance and a potential role of myeloid cells in the PD-L1<1% population. These findings shed light on potential next-generation therapies to overcome ICI resistance.Significance:ICI benefits patients with NSCLC, but resistance remains common. Our research highlights differences in tumor environments between primary and acquired resistance after ICI treatment, emphasizing distinct post-therapy approaches. Findings also suggest myeloid cells as key players in PD-L1–negative cases, guiding future treatment strategies to overcome resistance and improve outcomes.