Extracellular Matrix Composition in COPD Airways Changes Cell Fate of Airway Basal Stem Cells

Emily Liu; Avary Edwards; Bin Deng; Daniel Weiss; Amy Ryan

doi:10.1152/physiol.2026.41.S1.2247175

Abstract only Chronic obstructive pulmonary disease (COPD) is characterized not only by inflammatory and structural airway changes, but also by profound remodeling of the extracellular matrix (ECM). Although ECM abnormalities are a hallmark of COPD, their direct impact on airway epithelial regeneration, particularly on basal stem cell differentiation, remains poorly understood, representing a significant gap in knowledge regarding how the diseased microenvironment may perpetuate epithelial dysfunction. The objective of this study is to determine the impact of COPD-associated changes in ECM composition on airway basal cell differentiation. We hypothesized that lung-derived COPD ECM will drive basal cell differentiation in favor of mucus-secreting goblet cells. Human bronchial epithelial cells (HBECs) were expanded in PromoCell Airway Epithelial Cell Growth Medium and seeded onto transwell inserts for air–liquid interface (ALI) culture using PneumaCult ALI Maintenance Medium. ECM was generated from human airways by first decellularizing donor lungs via perfusion and then isolating specific regions of the airway by dissection; the decellularized tissue was then frozen, lyophilized, and then liquid nitrogen milled into a powder. ALI membranes were coated with lung-derived ECM, or PureCol as a control, and HBECs were differentiated for 28 days. Samples were collected for bulk RNA-sequencing and analyzed using standard bioinformatic pipelines and differential gene expression analysis (DESeq2). Immunohisto-/cyto-chemical staining was performed on paraffin-embedded cross-sections to assess differentiation and validate transcriptomic findings. In addition, en-face staining was performed on ALI cultures to determine the cell surface coverage of the respective functional epithelial cell types. HBECs differentiated on lung-derived ECM exhibited robust growth and differentiation comparable to PureCol controls, demonstrated by staining for basal (KRT5), goblet (MUC5AC), and ciliated (acetyl-α-tubulin) cell markers. However, HBECs differentiated on COPD-ECM showed a marked increase in goblet cells. This COPD ECM-driven secretory bias was more pronounced in female-derived BCs than in male-derived BCs. Notably, COPD-derived BCs grown on non-diseased ECM differentiated similarly to non-diseased controls, indicating that ECM composition, not BC intrinsic disease status, primarily drove the observed phenotypes. Interestingly, when HBECs with basal cell carcinoma were differentiated on non-diseased compared to COPD ECM, the basal cell hyperplasia was observed only on the COPD ECM, suggesting that the altered matrix composition in COPD affects the proliferation of HBECs. These findings demonstrate that HBECs can be functionally differentiated on lung-derived airway ECM at the ALI, and that the ECM composition in disease states impacts the growth and differentiation of HBECs. Together, these results indicate that ECM composition plays a critical role in airway epithelial regeneration and that, in COPD, disease-altered ECM may actively reinforce epithelial dysfunction and impaired repair. A greater understanding of ECM-cell interactions could have significant implications for tissue regeneration and cell-based therapies, which often do not account for pre-existing ECM abnormalities; healthy cells seeded onto diseased ECM may be unable to fully restore a damaged epithelium. This research was funded by NSF:ENG-BIOTECH #15265800 and the Iowa STEAD Family Foundation, awarded to ALR, NSF:ReCODE #2225554 awarded to ALR and DJW, NIH; NHLBI grant #1F31HL182238, awarded to ECL and the Vermont Biomedical Research Network Proteomics Facility is supported by NIH grant P20GM103449. This abstract was presented at the American Physiology Summit 2026 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.

Extracellular Matrix Composition in COPD Airways Changes Cell Fate of Airway Basal Stem Cells

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