Abstract 909: Single-cell transcriptomic profiling of non-small cell lung cancer uncovers inter- and intracell population structure across TCGA lung adenocarcinoma and lung squamous cancer subtypes

Abstract

Abstract Lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC) are the most prevalent types of non-all cell lung cancer (NSCLC), a leading cause of cancer death worldwide. In this study, we analyzed the transcriptomes of ~45,000 single cells (scRNA) from 13 NSCLC patients, including 5 LUAD cases which were collected and profiled at our institution. To correlate genomes and transcriptomes we performed Whole-Genome Sequencing (WGS) on 3 of these 5 LUAD cases. By comparing tumor tissue with matched adjacent non-malignant lung tissue we are able to confidently distinguish 13 cell-type specific clusters that unambiguously match previously characterized lineages. We developed algorithms for the identification of malignant cells derived from tumor tissue through scRNA analysis of copy number alterations and single nucleotide variants (SNV). Joint analysis of WGS and scRNA confirmed an enrichment of tobacco-associated SNVs among malignant cells of the tumor. Stromal cell types demonstrated consistent expression patterns across cases, while malignant cells demonstrated both inter- and intra-tumoral heterogeneity in their expression of signatures related to GPCR signaling, 3’ UTR mediated translational regulation, and cell-cell junction organization. In particular, one case displayed a unique pattern of intra-tumoral heterogeneity, as a subset of malignant cells robustly express a marker of pulmonary neuroendocrine cells, CGRP. Employing immunohistochemistry, the spatial organization of these malignant cells is revealed to be mutually exclusive within the tumor microenvironment and overlapping in expression of clinical markers of small-cell lung cancer. Finally, we deconvolved bulk TCGA LUAD and LUSC gene expression samples and analyzed the relationship between cell type specific gene expression in cell types of the lung and passenger mutation topographies. Our results provide insight into the molecular and clinical correlates of deconvolved NSCLC transcriptomes and provide a novel methodology with which to explore genomic variation at a single cell resolution. Furthermore, our dataset provides a resource for illuminating cancer-cell transcriptional changes and revealing key molecular drivers of tumor-stromal interactions in lung cancer. Citation Format: Kofi E. Gyan, Aditya Deshpande, Shaham Beg, Huasong Tian, Joel Rosiene, Marlon Stoeckius, Peter Smibert, Davide Risso, Juan Miguel Mosquera, Marcin Imielinski. Single-cell transcriptomic profiling of non-small cell lung cancer uncovers inter- and intracell population structure across TCGA lung adenocarcinoma and lung squamous cancer subtypes [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 909.