Eldredge, L. C. | bioRxiv Science

bioRxiv2025

Infants who develop BPD have an airway endotype defined by vimentin expression and ciliary loss

RationaleBronchopulmonary Dysplasia (BPD) results from abnormal lung development after preterm birth, with structural deficits at every respiratory tree level. BPD with lower airway disease is emerging as a clinically significant phenotype with increased mortality, and there is a significant knowledge gap in the molecular mechanisms whereby preterm birth disrupts normal airway development. ObjectivesTo develop a human model of lower airway disease after preterm birth and to characterize a molecular endotype of evolving BPD (eBPD) at baseline and in response to injury. MethodsWe used a combination of an ex vivo organotypic Airway Epithelial Cell (AEC models) and well-characterized pathologic and transcriptomic patient samples for quantitative immunohistochemistry and RNA sequencing analyses. Measurements and Main ResultsCompared to AECs from healthy patients, eBPD- derived AECs have a molecular endotype of reduced proliferation, impaired differentiation to ciliated epithelium, and an expanded vimentin-positive population with a transcriptional shift toward stromal cell-associated genes. With hyperoxia exposure, eBPD-derived AECs exhibited a pronounced vimentin response ex vivo, which parallels the increased vimentin expression of airway cells observed in lung tissue from human infants with BPD. ConclusionsIn this organotypic model of neonatal airway differentiation, we find that infants with eBPD have impaired differentiation, increased expression of vimentin, and concomitant loss of cilia, with an exaggerated increase in vimentin expression after hyperoxia injury, findings that mimic the effects of prematurity in airway cells in human patients. These data provide a foundation for future mechanistic studies interrogating the role of intermediate filaments in epithelial differentiation and repair.

cell biology↗

bioRxiv2025

A spatial transcriptomic atlas of acute neonatal lung injury across development and disease severity

A molecular understanding of lung organogenesis requires delineation of the timing and regulation of the cellular transitions that ultimately form and support a surface capable of gas exchange. While the advent of single-cell transcriptomics has allowed for the discovery and identification of transcriptionally distinct cell populations present during lung development, the spatiotemporal dynamics of these transcriptional shifts remain undefined. With imaging-based spatial transcriptomics, we analyzed the gene expression patterns in 17 human infant lungs at varying stages of development and injury, creating a spatial transcriptomic atlas of [~]1.2 million cells. We applied computational clustering approaches to identify shared molecular patterns among this cohort, informing how tissue architecture and molecular spatial relationships are coordinated during development and disrupted in disease. Recognizing that all preterm birth represents an injury to the developing lung, we created a simplified classification scheme that relies upon the routinely collected objective measures of gestational age and life span. Within this framework, we have identified cell type patterns across gestational age and life span variables that would likely be overlooked when using the conventional "disease vs. control" binary comparison. Together, these data represent an open resource for the lung research community, supporting discovery-based inquiry and identification of targetable molecular mechanisms in both normal and arrested human lung development.

genomics↗