Interleukin-13 disrupts type 2 pneumocyte stem cell activity
Kristen M. Glisinski, Adam J. Schlobohm, Sarah V. Paramore, Anastasiya Birukova, M. Arthur Moseley, Matthew W. Foster, Christina E. Barkauskas
Kristen M. Glisinski, Adam J. Schlobohm, Sarah V. Paramore, Anastasiya Birukova, M. Arthur Moseley, Matthew W. Foster, Christina E. Barkauskas
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Research Article Pulmonology Stem cells

Interleukin-13 disrupts type 2 pneumocyte stem cell activity

Abstract

The T helper 2 (Th2) inflammatory cytokine interleukin-13 (IL-13) has been associated with both obstructive and fibrotic lung diseases; however, its specific effect on the epithelial stem cells in the gas exchange compartment of the lung (alveolar space) has not been explored. Here, we used in vivo lung models of homeostasis and repair, ex vivo organoid platforms, and potentially novel quantitative proteomic techniques to show that IL-13 disrupts the self-renewal and differentiation of both murine and human type 2 alveolar epithelial cells (AEC2s). Significantly, we find that IL-13 promotes ectopic expression of markers typically associated with bronchiolar airway cells and commonly seen in the alveolar region of lung tissue from patients with idiopathic pulmonary fibrosis. Furthermore, we identify a number of proteins that are differentially secreted by AEC2s in response to IL-13 and may provide biomarkers to identify subsets of patients with pulmonary disease driven by “Th2-high” biology.

Authors

Kristen M. Glisinski, Adam J. Schlobohm, Sarah V. Paramore, Anastasiya Birukova, M. Arthur Moseley, Matthew W. Foster, Christina E. Barkauskas

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Figure 2

AEC2-derived organoids grown with IL-13 display altered morphology.

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AEC2-derived organoids grown with IL-13 display altered morphology. (A) ...
(A) 3D organoid cultures containing lineage-labeled AEC2s and PdgfraGFP+ stromal cells were grown with and without IL-13. Organoids grown with IL-13 appear larger and less dense. This morphology is observed with treatment with another Th2 cytokine, IL-4, as well. (B) When compared with control organoids, IL-13–treated organoids display lower colony-forming efficiency (CFE) and higher average diameter. Paired t tests; error bars indicate mean ± SD. (C) IL-13–treated spheres lack AEC1 staining (PDPN). Organoids treated with IL-4 display similar histological morphology. (D and F) Lineage-labeled AEC2s were isolated from WT SftpcTm mice (SpcTm WT) or from Stat6–/– SftpcTm mice (SpcTm Stat6–/–). PdgfraGFP+ stromal cells were isolated from WT PdgfraGFP mice or from Stat6–/– PdgfraGFP mice (PGFP Stat6–/–). These cells were plated in organoid culture in different combinations and grown to day 14. (D and E) Organoids derived from WT lineage-labeled AEC2s grow normally in control conditions, even if STAT6 signaling is abolished in the stroma. When IL-13 is added to WT organoids supported by either WT or Stat6–/– stromal cells, the typical IL-13 effect is observed. (F and G) Stat6–/– organoids grown with IL-13 do not display an increase in average sphere diameter, and the morphology appears grossly normal. These data suggest that the IL-13 effect is due to STAT6 signaling in the epithelium. One-way ANOVA; error bars indicate mean ± SD. Scale bars: 500 μm (A), 100 μm (C), and 500 μm (D). *P < 0.05; **P < 0.005.