Bulk transcriptional profiling of murine lung epithelial cells after expression of oncogenic KRAS. 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. Overall design: 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.