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 5

Proteomic analysis.

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Proteomic analysis. (A) Schematic. For cell-based proteomic analysis, li...
(A) Schematic. For cell-based proteomic analysis, lineage-labeled cells were dissociated from Matrigel, and protein isolated from them was trypsinized and peptides analyzed by nanoflow liquid chromatography–tandem mass spectrometry (nano–LC-MS/MS). For secretomic analysis, conditioned medium was collected at day 14 and protein trypsinized, labeled, fractionated, and analyzed by LC-MS/MS. (B) Heatmap showing differential expression of proteins in lineage-labeled epithelial cells from control and IL-13–exposed cultures. Proteins depicted are significant at P < 0.05 and fold change > ±1.5 in IL-13 vs. control. (C) Three representative differentially expressed proteins from cell-based proteomic analysis in B. Chitinase-like protein A (CHIA) and collagen α 1(XIV) chain (COL14A1) were significantly upregulated in AEC2-derived cells in IL-13–treated cultures compared with controls. Conversely, there was less expression of fatty acid binding protein 5 (FABP5) in AEC2-derived cells from IL-13–treated cultures compared with controls. CHIA WT, IL-13 vs. control, P = 0.001965; CHIA STAT6, IL-13 vs. control, P = 0.005662; COL14A1 WT, IL-13 vs. control, P = 0.011875; COL14A1 STAT6, IL-13 vs. control, P = 0.0.00319; FABP5 WT, IL-13 vs. control, P = 0.015991; FABP5 STAT6, IL-13 vs. control, P = 0.0.004382. *P < 0.05; **P < 0.005; paired, 2-tailed t test. (D) Volcano plot depicting proteins that are differentially expressed in the secretome of cultures treated with IL-13 versus control. Error bars represent mean ± SD. Analysis done with 3 biological replicates for each condition.