D1.489 - The establishment and characteristics of type 2 inflammation in nasal organoids

Type 2 inflammation plays a central role in upper airway diseases such as chronic rhinosinusitis, with the role of interleukin-13 (IL-13), recognized as a key driver of epithelial barrier dysfunction and airway remodeling. However, the lack of robust and physiologically relevant human in vitro models has limited mechanistic studies and therapeutic evaluation. This study aimed to establish and characterize a human nasal organoid model of type 2 inflammation to investigate IL-13 induced epithelial injury and differentiation changes.

Human nasal epithelial cells (HNECs) were expanded and embedded in growth factor reduced extracellular matrix to generate three-dimensional nasal organoids. Organoid growth and morphology were monitored over time. Acute and chronic injury models were established using varying concentrations of IL-13. Epithelial barrier integrity was assessed by FITC permeability assays. Tight junction proteins (ZO-1 and occludin) and epithelial cell lineage markers, including acetylated α-tubulin (ciliated cells) and MUC5AC (goblet cells) were evaluated by immunofluorescence staining. Gene expression of basal cell, ciliated cell, goblet cell, and barrier-related markers was quantified by RT-qPCR.

Stable nasal organoids with well-defined luminal structures were successfully established. Acute IL-13 exposure induced a dose-dependent increase in epithelial permeability, indicating barrier dysfunction. In the chronic injury model, low-dose IL-13 (2ng/mL) consistently produced a stable injury phenotype without significant cell death. IL-13 treatment disrupted tight junction organization and altered the expression of ZO-1 and Occludin at both protein and mRNA levels. Furthermore, IL-13 promoted goblet cell differentiation, while suppressing ciliated cell development, accompanied by increased MUC5AC expression and reduced FOXJ1 expression, reflecting epithelial remodeling characteristic of type 2 inflammation.

We established a reproducible human nasal organoid model that effectively recapitulates IL-13 mediated type 2 inflammatory features, including epithelial barrier impairment and altered cellular differentiation. This model provides a valuable platform for mechanistic studies of upper airway inflammation and for evaluating potential therapeutic strategies targeting epithelial barrier protection.