598 - Single-cell Transcriptional Profiling of Mouse lung Following Hyperoxia-induced Injury

Poster Number: 598
Publication Number: 598.1597

Background: Hyperoxia disrupts lung development in mice and causes bronchopulmonary dysplasia (BPD) in neonates. Our understanding of cell-specific gene expression changes that drive hyperoxia-induced lung damage is still evolving.

Objective: We assessed mouse lung using single cell RNA sequencing (scRNA-seq) changes in an established mouse model of hyperoxia-induced acute lung injury (HALI).

Design/Methods: Newborn mouse pups were randomized to room air or to HALI using the following established model - exposure to 80-85% O2 from postnatal (PN) day 1-3, PN1-7, PN1-14, or PN1-21. We harvested lung tissue on PN 3, 7, 14 and 21 after hyperoxia exposure, for histology (n=6/group) and 62520 cells from lungs for scRNA-seq at PN14 (n=2/group). Alveolarization was quantified on lung sections by radial alveolar count, mean alveolar diameter and alveolar septal thickness. Chromium 10× Genomics library and sequencing were performed on single-cell suspensions of lung tissues. Final libraries were sequenced on the Illumina NovaSeq 6000 sequencing system (paired-end multiplexing run, 150bp) by LC-Bio Technology Co., Ltd. (Hangzhou, China) at a minimum depth of 20,000 reads per cell.

Results: There was a decreased number of alveoli, increased alveolar size and thickening of alveolar wall in the hyperoxia groups at days 14 and 21 (Figure 1A-C). Eighteen cell types were identified. Compared with control mice, there was no significant difference in cell composition between the HALI model and control groups at 14 days (Figure 1D-E, Table 1). However, expression of multiple pro-fibrotic, and anti-angiogenic genes was significantly increased, while expression of sftpa1, sftpb and sftpc was down-regulated in HALI group.

Conclusion(s): Several features of HALI were recapitulated in our mouse model of hyperoxia-induced lung injury. Single-cell transcriptional profiling implicate pro-fibrotic and anti-angiogenic genes in the pathogenesis of HALI and may be targeted for therapeutic considerations.

Funding Sources: This work was supported by grants from the National Natural Science Foundation of China (No.81873845).