Am J Respir Crit Care Med

. 2023 Feb 2.

 doi: 10.1164/rccm.202205-0953OC. Online ahead of print. https://pubmed.ncbi.nlm.nih.gov/36731066/

A Tracheal Aspirate-Derived Airway Basal Cell Model Reveals a Proinflammatory Epithelial Defect in Congenital Diaphragmatic Hernia

Richard Wagner ¹, Gaurang M Amonkar ², Wei Wang ², Jessica E Shui ², Kamakshi Bankoti ², Wai Hei Tse ³, Frances A High ^4 5, Jill M Zalieckas ⁶, Terry L Buchmiller ⁵, Augusto Zani ^7 8 9, Richard Keijzer ¹⁰, Patricia K Donahoe ⁴, Paul Hubert Lerou ², Xingbin Ai ¹¹

Affiliations expand

• PMID: 36731066
• DOI: 10.1164/rccm.202205-0953OC

Abstract

Rationale: Congenital diaphragmatic hernia (CDH) is characterized by incomplete closure of the diaphragm and lung hypoplasia. The pathophysiology of lung defects in CDH is poorly understood.

Objectives: To establish a translational model of human airway epithelium in CDH for pathogenic investigation and therapeutic testing.

Methods: We developed a robust methodology of epithelial progenitor derivation from tracheal aspirates of newborns. Basal stem cells (BSCs) from CDH patients and preterm and term, non-CDH controls were derived and analyzed by bulk RNA-sequencing, ATAC-sequencing, and air-liquid-interface differentiation. Lung sections from fetal human CDH samples and the nitrofen rat model of CDH were subjected to histological assessment of epithelial defects. Therapeutics to restore epithelial differentiation were evaluated in human epithelial cell culture and the nitrofen rat model of CDH.

Measurements and main results: Transcriptomic and epigenetic profiling of CDH and control BSCs reveals a proinflammatory signature that is manifested by hyperactive NF-κB and independent of severity and hernia size. In addition, CDH BSCs exhibit defective epithelial differentiation in vitro that recapitulates epithelial phenotypes found in fetal human CDH lung samples and fetal tracheas of the nitrofen rat model of CDH. Furthermore, blockade of NF-κB hyperactivity normalizes epithelial differentiation phenotypes of human CDH BSCs in vitro and in nitrofen rat tracheas in vivo.

Conclusions: Our findings have identified an underlying proinflammatory signature and BSC differentiation defects as a potential therapeutic target for airway epithelial defects in CDH.

Keywords: Congenital Diaphragmatic Hernia; Inflammatory gene expression signature; Lung Hypoplasia; Nitrofen Rat Model; Tracheal Aspirate derived lung progenitor cells.