Natural killer cell TGF-β signaling regulates senolytic activity and vascular patterning in the postnatal lung

Abstract Background Bronchopulmonary dysplasia (BPD) is a disease of neonatal lung development that is linked to impaired pulmonary vascularization, dysregulated transforming growth factor-β (TGF-β) signaling and the accumulation of senescent cells. Despite the established role for TGF-β signaling in promoting vascular remodeling and suppressing the senolytic activity of natural killer (NK) cells, the contribution of NK cell TGF-β signaling to postnatal lung patterning and the pathogenesis of BPD remains unclear. Methods Mice bearing an NK cell-selective deletion of the type-II TGF-β receptor ( Tgfbr2 NK-/- ) were analyzed for vascular and alveolar structure, lung NK cell infiltration, senescence markers and lung function testing across neonatal and adult timepoints. Single-cell RNA sequencing of lung tissue from both neonatal mice and human infants with BPD was performed. The effect of enhanced NK cell activity in a hyperoxia-induced model of BPD was assessed in Tgfbr2 NK-/- neonates, as well as pharmacologically, using the TGF-β ligand trap/IL-15 superagonist, HCW9218. Results Neonatal Tgfbr2 NK-/- mice exhibited a baseline reduction in distal arteriolar density, impaired alveolarization, and sex-specific deficits in long-term lung function. Single-cell RNA sequencing identified the excessive clearance of senescent endothelial cells by TGF-β insensitive NK cells in the lungs of Tgfbr2 NK-/- neonates, which served as a contributor of the BPD-like phenotype observed in naïve animals. Tgfbr2 NK-/- mice were protected from impaired lung development in the hyperoxia model. Sequencing from lung tissue from infants with BPD confirmed excessive TGF-β signaling and cytotoxic impairment in NK cells. Treatment with HCW9218 prevented senescent cell accumulation and rescued lung development in the hyperoxia mouse model. Conclusions These findings identify TGF-β as a tunable regulator of NK cell senolytic activity that is essential to normal postnatal lung development. Excessive NK cell TGF-β signaling contributes to impaired lung development following exposure to neonatal hyperoxia and may serve as a viable therapeutic target for human BPD.