Single-cell analysis of tobacco-associated lung adenocarcinoma development

Understanding cellular processes underlying early lung adenocarcinoma (LUAD) development is needed to devise intervention strategies. Here, we performed single-cell RNA sequencing (scRNA-seq) of mouse lungs from Gprc5a-/- mice during lung tumor development. We coupled scRNA-seq analysis with spatial transcriptomics of tumor-bearing lungs. scRNA-seq profiling was performed on lungs from Gprc5a-/- mice treated with nicotine-derived nitrosamine ketone (NNK) or saline at the end of exposure (EOE, timepoint at which lungs are still clear of lesions), at 3 months and at 7 months post-exposure, the time point of Kras-mutant LUAD onset in all mice (n = 4 mice per group and time point). We also performed scRNA-seq of GFP+ and GFP- fractions of lungs from Gprc5a-/- mice carrying alveolar type II reporter (Gprc5a-/-; SftpcCreER/+; RosaSun1GFP/+) at three months post-exposure to saline of NNK (n = 2 mice per group). scRNA-seq was performed using the 10X Genomics platform. Spatial transcriptomics of tumor-bearing lungs from Gprc5a-/- mice at 7 months post-exposure to NNK (n = 3) was performed using the Visium platform from 10X Genomics to further characterize and validate scRNA-seq findings in a spatial context.