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 4

AEC2s that coexpress SCGB1A1 are more capable of growth in the presence of IL-13 than AEC2s that do not express SCGB1A1.

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AEC2s that coexpress SCGB1A1 are more capable of growth in the presence ...
(A) Lungs were isolated from Scgb1a1CreER/+ Tm/+ (Scgb1a1Tm) mice following TMX administration. Airway cells and scattered alveolar epithelial cells (white arrowheads) are labeled in these mice; rare BASCs (SFTPC+SCGB1A1+ cells located in the terminal bronchiolar region; yellow arrowhead) are labeled. Lungs were dissociated and a single-cell suspension was stained for LysoTracker (LT) in order to mark AEC2s. Both Tomato+ (Sgcb1a1–lineage-labeled) LT+ cells (AEC2s that coexpress SCGB1A1; gray box) and Tomato– (non–lineage-labeled) LT+ cells (AEC2s that do not coexpress SCGB1A1; magenta box) were sorted via FACS and cultured in equal numbers with PGFP+ stromal support cells for 14 days with and without IL-13. (B and C) The CFE of Tomato+ Scgb1a1–lineage-labeled AEC2s increases with IL-13 exposure (calculated based on fold change in CFE from control). The baseline CFE of Scgb1a1Tm LT+ control cultures is 3.58%. This CFE increases to 5.25% in the presence of IL-13. The baseline CFE of non–lineage-labeled AEC2s that are LT+ is 14.42% in control conditions. This CFE drops to 11.68% in the presence of IL-13. The average sphere diameter of IL-13–treated organoids derived from Tomato+ Scgb1a1-expressing AEC2s is higher than IL-13–treated organoids derived from Tomato– Scgb1a1 lineage– AEC2s. (D) There is notable KRT5 staining in organoids derived from Tomato+ Scgb1a1–lineage-labeled, LT+ AEC2s compared with control spheres. One-way ANOVA with multiple comparisons; error bars represent mean ± SD. Scale bars: 100 μm (A), 500 μm (B), and 100 μm (D). *P < 0.05; **P < 0.005.