Single-cell RNA Sequencing of Murine Lungs to Compare Two Different Models of Alveolar Simplification

Neonatal hyperoxia exposure causes alveolar simplification in mice and has been employed as a model to study bronchopulomonary dysplasia, or chronic lung disease of prematurity. Loss of epithelial TGF-beta signaling has also been shown to cause alveolar simplification in mouse models of lung development. We used single cell RNA sequencing (scRNA-seq) to analyze both of these models of lung injury to identify shared molecular and cellular mechanisms that contribute to pathogenic alveolar simplification. Neonatal mice on the C57BL/6 background were exposed to either hyperoxia (75% oxygen) or normoxia for 10 days begnning on day 0 and were recovered in normoxia until harvest at day 14. Tgfbr2 flox/flox (WT) mice were compared to Nkx2.1-cre;Tgfbr2 flox/flox (KO) littermates in either normoxic (RA) or hyperoxic (O2) conditions. Two mice of each genotype and of each exposure were harvested on day 7 and day 14 and lung tissues were digested to generate a single cell suspension and analyzed using scRNAseq.