Abstract 3393: Altered immune response in the transcriptome of patients with lung cancer
Authors: Hijazi Kahkeshan , Lel Julian , Billatos Ehab , Moses Elizabeth , Stevenson Christopher S. , Lorenzi Matthew V. , Liu Gang , Campbell Joshua D. , Koga Yusuke , Zhang Jiarui , Duan Fenghai , Marques Helga , Lenburg Marc E. , Spira Avrum E. , Beane Jennifer 
Published: August 2019

Introduction The immune system is critical to surveying and eradicating abnormal cells, but tumor cells develop ways to escape immunosurveillance and induce an immunosuppressive state. We previously developed and validated gene expression (GE) signatures measured in the normal airway-epithelial brushings in patients undergoing bronchoscopy for suspicion of lung cancer (LC). In this study, we seek to understand if the immunosuppressive environment extends to the airway field of injury via profiling of endobronchial biopsies from the central airway containing a broader range of cell types, including immune cells.

Methods Endobronchial biopsies from normal-appearing regions of the central airway were collected from ever smokers undergoing workup of indeterminate pulmonary nodules (7-30 mm in diameter) suspicious for LC at military and VA hospitals within the DECAMP consortium. Initially, total RNA from the biopsies (n=44, discovery-set) were isolated and sequenced. Reads were aligned to hg19 using STAR and gene level counts were quantified with RSEM. Poor quality samples were removed using FASTQC and RSEQC. Differential GE associated with cancer status was identified using edgeR, adjusting for smoking-status, COPD and sample quality. RNA from additional endobronchial biopsies (n=49, validation-set) were isolated and preprocessed similarly. Genes differentially expressed with LC status in the discovery-set were tested in the validation-set using gene set variation analysis (GSVA). Functional enrichment of cancer associated genes was explored using Enrichr. Comparison of cancer signatures identified in previously published LC studies was investigated using GSEA. CIBERSORT, xCell, TIMER software and single-cell RNA sequencing data generated from airway brushings were used to deconvolute the immune cell content of the bulk biopsy samples.

Results We identified a GE signature associated with LC which was significantly and concordantly enriched in the validation set of biopsies and two previously published studies of LC-associated GE in airway brushings. Genes decreased in LC patient biopsies were enriched for genes involved in immune-related pathways, including cytokine interactions, the inflammatory response and neutrophil degranulation. Computational deconvolution and comparison with single-cell RNAseq data predicts a decrease in neutrophils in the airway of LC patients.

Conclusion We identified LC-associated GE alterations in smokers presenting with indeterminate pulmonary nodules. Down-regulated genes in LC subjects are strongly associated with immune system function, specifically neutrophil biology. Subjects with LC appear to have an immunosuppressive environment directed towards myeloid cell populations, and this could have implications for the future development of immunoprevention therapies.

Citation Format: Kahkeshan Hijazi, Julian Lel, Ehab Billatos, Elizabeth Moses, Christopher S. Stevenson, Matthew V. Lorenzi, Gang Liu, Joshua D. Campbell, Yusuke Koga, Jiarui Zhang, Fenghai Duan, Helga Marques, Marc E. Lenburg, Avrum E. Spira, Jennifer Beane. Altered immune response in the transcriptome of patients with lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3393.