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HNEC-derived iPSCs are transcriptionally equivalent to iPSCs generated from other somatic sources

ABSTRACT: Studying epithelial dysfunction in respiratory diseases requires physiologically relevant human models that faithfully recapitulate airway structure and function. Primary human nasal epithelial cells (hNECs) represent an accessible and clinically validated system, widely used for mutation-specific drug theratyping. However, the broader potential of hNECs as an integrated experimental platform remains underexplored. Here, we establish a customized hNEC-based platform spanning ex vivo, in vitro, and stem cell-based applications, with direct relevance to precision medicine. Using freshly fixed nasal brushings, we developed a quantitative single-cell metric to assess Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) localization at the ionocyte plasma membrane, identifying a candidate ex vivo biomarker sensitive to disease status and pharmacological modulation in cystic fibrosis (CF). In parallel, transcriptomic profiling demonstrated that air-liquid interface-differentiated hNECs closely recapitulate the molecular features of native nasal epithelium. Functionally, differentiated hNECs reproduced key properties of airway surface liquid (ASL) physiology and its dysregulation under inflammatory and CF conditions. Finally, we show that basal hNECs provide a robust and accessible source for induced pluripotent stem cell (iPSC) generation. hNEC-derived iPSCs exhibited canonical pluripotency features, differentiated efficiently into germ layer derivatives and lung progenitors, and displayed transcriptomic profiles indistinguishable from iPSCs generated from other somatic sources. Together, these findings position hNECs as a versatile, patient-specific platform enabling ex vivo biomarker discovery, in vitro modeling of airway physiology, and generation of stem cell–based models, thereby expanding their utility for respiratory research and precision medicine.

ORGANISM(S): Homo sapiens

PROVIDER: GSE319532 | GEO | 2026/08/27

REPOSITORIES: GEO

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Project description:E2 exposure significantly decreased peak viral titer in hNECs from female donors. We used microarray analyses to identify global gene expression patterns between E2 and vehicle exposed hNECs from female donors Influenza causes an acute infection characterized by virus replication in respiratory epithelial cells. The severity of influenza and other respiratory diseases changes over the life course and during pregnancy in women, suggesting that sex steroid hormones, such as estrogens, may be involved. Using primary, differentiated human nasal epithelial cell (hNEC) cultures from adult male and female donors, we exposed cultures to the endogenous 17Î²-estradiol (E2) or select estrogen receptor modulators (SERMs), then infected cultures with a seasonal influenza A virus (IAV) to determine whether estrogenic signaling could affect the outcome of IAV infection and whether these effects where sex-dependent. Estradiol, raloxifene, and bisphenol A decreased IAV titers in hNECs from female, but not male, donors. The estrogenic decrease in viral titer was dependent on the genomic estrogen receptor- 2 (ESR2) as neither genomic ESR1 nor non- genomic GPR30 were expressed in hNEC cultures and addition of the genomic ER antagonist ICI 182,780 reversed the antiviral effects of E2. Treatment of hNECs with E2 had no effect on interferon or chemokine secretion, but significantly downregulated cell metabolic processes, including genes that encode for zinc finger proteins, many of which contain estrogen response elements in their promoters. These data provide novel insights into the cellular and molecular mechanisms of how natural and synthetic estrogens impact IAV infection in respiratory epithelial cells derived from humans. Primary human nasal epithelial cells from females were exposed to E2 for 24h prior to infection, then infected with an H3N2 strain of influenza a virus for 2 hours. At 24 and 48h post infection, hNECs were collected in Trizol for RNA extraction and hybridization on Affymetrix Human Gene ST 2.0 microarrays.

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HNEC-derived iPSCs are transcriptionally equivalent to iPSCs generated from other somatic sources

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