Pediatr Surg Int

. 2022 Mar 2. doi: 10.1007/s00383-022-05096-0. Online ahead of print. https://pubmed.ncbi.nlm.nih.gov/35235015/

Tenascin C is dysregulated in hypoplastic lungs of miR-200b ^-/- mice

Moritz Markel ^1 2, Wai Hei Tse ¹, Nolan DeLeon ¹, Daywin Patel ¹, Shana Kahnamouizadeh ¹, Martin Lacher ², Richard Wagner ², Richard Keijzer ³Affiliations expand

• PMID: 35235015
• DOI: 10.1007/s00383-022-05096-0

Abstract

Purpose: We previously demonstrated that absence of miR-200b results in abnormal lung development in congenital diaphragmatic hernia due to imbalance between epithelial and mesenchymal cells. Tenascin C is a highly conserved extracellular matrix protein involved in epithelial to mesenchymal transition, tissue regeneration and lung development. Considering the involvement of Tenascin C and miR-200b and their potential interaction, we aimed to study Tenascin C during lung development in the absence of miR-200b.

Methods: We collected lungs of miR-200b^-/- mice (male, 8 weeks). We performed Western blot (WB) analysis (N = 6) and immunofluorescence (N = 5) for Tenascin C and alpha smooth muscle actin and RT-qPCR for Tenascin C gene expression (N = 4).

Results: Using WB analysis, we observed a decreased total protein abundance of Tenascin C in miR-200b^-/- lungs (miR-200b^+/+: 3.8 × 10⁷ ± 1 × 10⁷; miR-200b^-/-: 1.9 × 10⁷ ± 5 × 10⁶; p = 0.002). Immunofluorescence confirmed decreased total Tenascin C in miR-200b^-/- lungs. Tenascin C was significantly decreased in the mesenchyme but relatively increased in the airways of mutant lungs. Total lung RNA expression of Tenascin C was higher in miR-200b^-/- lungs.

Conclusion: We report dysregulation of Tenascin C in lungs of miR-200b^-/- mice. This suggests that absence of miR-200b results in abnormal Tenascin C abundance contributing to the lung hypoplasia observed in miR-200b^-/- mice.

Keywords: Congenital diaphragmatic hernia; Pulmonary hypoplasia; Tenascin C; miR-200b.

© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.