Bronchopulmonary Dysplasia with Pulmonary Hypertension Associates with Loss of Semaphorin Signaling and Functional Decrease in FOXF1 Expression

Abstract Lung injury in preterm infants leads to structural and functional respiratory deficits, with a risk for bronchopulmonary dysplasia (BPD) that in its most severe form is accompanied by pulmonary hypertension (PH). To examine cellular and molecular dynamics driving evolving BPD in humans, we performed single-cell RNA sequencing of preterm infant lungs in early stages of BPD and BPD+PH compared to term infants. Analysis of the endothelium revealed a unique aberrant capillary cell-state primarily in BPD+PH marked by ANKRD1 expression. Predictive signaling analysis identified deficits in the semaphorin guidance-cue signaling pathway and decreased expression of pro-angiogenic transcription factor FOXF1 within the alveolar parenchyma in neonatal lung samples with BPD/BPD+PH. Loss of semaphorin signaling was replicated in a murine BPD model and in humans with alveolar capillary dysplasia (ACDMPV), suggesting a mechanistic link between the developmental programs underlying BPD and ACDMPV and a critical role for semaphorin signaling in normal lung development. Competing Interest Statement Disclosures: JAK reports grant funding from Boehringer Ingleheim and Bristol-Myers Squibb outside the scope of this work, and serves on the Scientific Advisory Board for APIE and ARDA, outside of the scope of this work. Footnotes Disclosures: JAK reports grant funding from Boehringer Ingelheim and Bristol-Myers Squibb outside the scope of this work, and serves on the Scientific Advisory Board for APIE and ARDA, outside of the scope of this work. Revision reflects addition of references to discussion and text edits for clarity.