Abstract 206: Proteogenomic classifications and outcome in squamous cell carcinoma of the lung
Authors: Slebos Robbert J.C. , Stewart Paul A. , Welsh Eric A. , Cen Ling , Zhang Yonghong , Chen Zhihua , Cheng Chia-Ho , Pettersson Fredrik , Berglund Anders E. , Zhang Guolin , Fang Bin , Izumi Victoria , Yoder Sean J. , Fellows Katherine M. , Chen Y Ann , Teer Jamie , Eschrich Steven , Koomen John , Haura Eric 
Published: July 2017

Introduction: Genomic analyses have yielded a tremendous amount of knowledge on the genetic changes in lung cancers, but translating this information into actionable data benefitting patients has proven difficult. The integration of proteomic analyses with genomics and gene expression profiling allows a more detailed description of the biological processes, thus improving our understanding of cancer phenotypes. These insights can potentially be used for better classification and help to guide patient selection for targeted therapies.

Experimental: We analyzed 116 surgically resected squamous cell lung carcinoma samples for copy-number alterations, gene expression profiling, targeted exome-sequencing and global proteomic profiling. The cohort consisted of mostly early stage tumors (83% Stage I or II) with complete follow-up (median 58 months). Copy number status was analyzed using the Affymetrix CytoScan array, DNA mutation status was assessed using a customized version of the Agilent Comprehensive Cancer Panel for targeted sequencing, and gene expression profiling was carried out by RNA-sequencing. Proteomic analysis was performed using TMT labeling, 12- fraction bRPLC separation and LC-MS/MS analysis with a Thermo Q-Exactive mass spectrometer. Database searches were performed using multiple search engines against RefSeq version 78, and summarized using IDPicker 3.

Results: The non-redundant protein inventory consisted of more than 6,000 protein groups with a protein FDR &lt;5%. Tumors were classified according to well established gene expression criteria into 4 classes: Classical, Basal, Primitive and Secretory. At the protein level, the Classical subtype was associated with xenobiotic and energy metabolism; the Basal subtype with defense responses and extracellular matrix changes; the Primitive subtype with nucleic acid metabolism; and the Secretory subtype with p38 signaling. These findings confirm and expand on previous mRNA expression studies of squamous cell lung carcinoma. Proteomics-based classification identified two sub-categories within the Classical subtype, which were characterized by inflammatory and stress response signaling. Within this group, patients with high expression of inflammation-associated proteins had better cancer-specific survival than those with low expression (p=0.04, Log-rank test). Targeted exome sequencing of 154 cancer-associated genes revealed frequent mutations in TP53, CDKN2A, NFE2L2, and other genes. Proteomic expression of genes located in amplified chromosomal regions was used to identify driver genes in squamous cell lung carcinoma.

Conclusion: Our results provide new biological insights from the addition of protein measurements to genomic datasets that have the potential to improve classification. The data suggest that proteins involved in immune responses are important for the biological behavior and outcome of the Classical subtype in squamous cell lung carcinoma.

Citation Format: Robbert J.C. Slebos, Paul A. Stewart, Eric A. Welsh, Ling Cen, Yonghong Zhang, Zhihua Chen, Chia-Ho Cheng, Fredrik Pettersson, Anders E. Berglund, Guolin Zhang, Bin Fang, Victoria Izumi, Sean J. Yoder, Katherine M. Fellows, Y Ann Chen, Jamie Teer, Steven Eschrich, John Koomen, Eric Haura. Proteogenomic classifications and outcome in squamous cell carcinoma of the lung [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 206. doi:10.1158/1538-7445.AM2017-206