Abstract 5572: A COX-2-based pro-tumourigenic inflammatory signature predicts poor outcome in early-stage non-small cell lung cancer
Authors: Fife Victoria , Roberts Matthew , Bromley Christian P. , Morgan Derrick , Atkinson Sophie , Zhou Cong , Chaturvedi Anshuman , Bagley Steven , Ashton Garry , Coussens Lisa M. , Crosbie Philip A. , Dive Caroline , Kilgour Elaine , Zelenay Santiago 
Published: June 2023

Abstract
Lung cancer is the most common cause of cancer-related deaths with Non-Small Cell Lung Cancer (NSCLC) accounting for ~85% of cases. Patients with early-stage NSCLC (stages I-IIIA) undergo surgery with curative intent but up to 30% relapse. Identification of biomarkers that predict outcome at the point of surgery could inform patient management. Here, we explored whether a COX-2-based pro-tumourigenic inflammatory gene signature (PTI), measured in surgically resected samples from early-stage NSCLC patients, can predict patient outcome. Gene expression analysis was performed on 30 formalin fixed paraffin embedded surgical tumour samples from early-stage NSCLC patients with 36-63 months follow-up, using a custom code-set on the NanoString nCounter® platform. Additionally, in silico analysis of early-stage NSCLC data from The Cancer Genome Atlas (TCGA) was undertaken. The PTI gene signature and a published signature associated with T cell-inflamed tumours measuring IFNγ activity (IFNG)1 were assessed in both datasets and correlated with patient outcome. NanoString gene expression data was robust, displaying good correlations with immunohistochemistry (IHC) expression for key proteins such as CD8 and COX-2, R=0.88, p&lt;1 × 10−6 and R=0.83, p&lt;1 × 10−6, respectively. The IFNG signature score, widely reported to associate with response to checkpoint inhibitors2, did not correlate with outcome and consistent with this observation neither did CD8 protein expression (by IHC). In contrast, patients with LUSC (n=16) with the highest (upper quartile) PTI score showed significantly worse overall survival (p&lt;0.01). Median overall survival for these PTIhigh patients was &lt;6 months vs 4.5 years for LUSC patients with PTIlow scores. Analysis of TCGA data from stage I-IIIA LUAD/LUSC cases confirmed these contrasting findings for the PTI compared with the IFNG signature. A multiplexed IHC panel for immune contexture analysis was successfully transferred from the Coussens Laboratory Oregon to CRUK Manchester3 and with inclusion of COX-2 in this panel and adaptation of the pipeline for use with the HALO image analysis platform, analysis of the immune contexture in PTI high versus low tumours is underway. Transcriptional profiling of early-stage NSCLC samples revealed that COX-2-associated inflammation predicts patient outcome following surgery. These data highlight the importance of pro-tumourigenic inflammation in early-stage NSCLC. Findings are under validation in a larger cohort, including early-stage NSCLC samples from Manchester’s early-detection screening studies and mapping of the immune contexture of PTI high versus low tumours is underway. 1: Ayers, M., et al., J Clin Invest., 2017 2: Danaher, P., et al., J Immunother Cancer., 2018 3: Banik, G., et al., Methods Enzymol., 2020
Citation Format: Victoria Fife, Matthew Roberts, Christian P. Bromley, Derrick Morgan, Sophie Atkinson, Cong Zhou, Anshuman Chaturvedi, Steven Bagley, Garry Ashton, Lisa M. Coussens, Philip A. Crosbie, Caroline Dive, Elaine Kilgour, Santiago Zelenay. A COX-2-based pro-tumourigenic inflammatory signature predicts poor outcome in early-stage non-small cell lung cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5572.