Bulk transcriptional profiling of murine lung epithelial cells after expression of oncogenic KRAS

The goal of this study was to define the transcriptional changes that follow oncogenic KRAS-G12D expression in aveolar type 2 (AT2) cells. We induced KRAS-G12D expression in AT2 cells in vitro and studied transcriptional changes using bulk RNA-Sequencing seven days after plating. Alveolar type 2 (AT2) cells were sorted using FACS (CD31-/CD45-/EPCAM+/SCA1-) from lungs of mixed and pooled male and female uninduced KRAS-LSL-G12D;LSL-YFP (KY) or KRAS-LSL-G12D; p53FL/FL; LSL-YFP (KPY) mice. AT2 cells were then not induced (NI control), or infected with Cre expressing (CRE), or empty (EMP control) adenovirus and plated in 3D organoid co-culture. Seven days after plating, multiple wells (>10) were pooled, single cell suspensions were made from the organoid cultures, and epithelial cells were FACS sorted using EPCAM. This experiment was repeated 4 times for 3-4 biological samples per condition.