The proportion of alveolar type 1 cells decreases in murine hypoplastic congenital diaphragmatic hernia lungs

PLoS One. 2019 Apr 17;14(4):e0214793. doi: 10.1371/journal.pone.0214793. eCollection 2019.

Abstract

Background: Pulmonary hypoplasia, characterized by incomplete alveolar development, remains a major cause of mortality and morbidity in congenital diaphragmatic hernia. Recently demonstrated to differentiate from a common bipotent progenitor during development, the two cell types that line the alveoli type 1 and type 2 alveolar cells have shown to alter their relative ratio in congenital diaphragmatic hernia lungs.

Objective: We used the nitrofen/bisdiamine mouse model to induce congenital diaphragmatic hernia and accurately assess the status of alveolar epithelial cell differentiation in relation to the common bipotent progenitors.

Study design: Pregnant Swiss mice were gavage-fed with nitrofen/bisdiamine or vehicle at embryonic day 8.5. The administered dose was optimized by assessing the survival, congenital diaphragmatic hernia and facial abnormality rates of the exposed mouse pups. NanoCT was performed on embryonic day 11.5 and 16.5 to assess the embryonic and early canalicular stages of lung development. At embryonic day 17.5 corresponding to late canalicular stage, congenital diaphragmatic hernia lungs were characterized by measuring the lung weight/body weight ratio, morphometry, epithelial cell marker gene expression levels and alveolar cell type quantification.

Results: Nitrofen/bisdiamine associated congenital diaphragmatic hernia lungs showed delayed development, hypoplasia with morphologic immaturity and thickened alveolar walls. Expression levels of distal epithelial progenitor marker Id2 increased, alveolar type 1 cell markers Pdpn and Hopx decreased, while type 2 cell markers pro-SPC and Muc1 remained constant during the canalicular stage. The number of Pdpn+ type 1 alveolar cells also decreased in congenital diaphragmatic hernia lungs.

Conclusion: The mouse nitrofen/bisdiamine model is a potential model allowing the study of congenital diaphragmatic hernia lung development from early stages using a wide array of methods. Based on this model, the alveolar epithelium showed a decrease in the number of alveolar type 1 cell in congenital diaphragmatic hernia lungs while type 2 cell population remains unchanged.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alveolar Epithelial Cells / metabolism
  • Alveolar Epithelial Cells / pathology*
  • Animals
  • Cell Count
  • Cell Differentiation
  • Diamines / toxicity
  • Disease Models, Animal
  • Embryonic Stem Cells / metabolism
  • Embryonic Stem Cells / pathology
  • Female
  • Hernias, Diaphragmatic, Congenital / chemically induced
  • Hernias, Diaphragmatic, Congenital / embryology
  • Hernias, Diaphragmatic, Congenital / pathology*
  • Lung / abnormalities*
  • Lung / embryology
  • Lung / pathology
  • Mice
  • Organ Size
  • Phenyl Ethers / toxicity
  • Pregnancy
  • Teratogens / toxicity

Substances

  • Diamines
  • Phenyl Ethers
  • Teratogens
  • N,N'-bis(dichloroacetyl)-1,8-octamethylenediamine
  • nitrofen

Grants and funding

T.M.N. was funded with support of the Erasmus+ Programme of the European Union (Framework Agreement number: 2013-0040). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. NanoCT scans were done using the NanoTom which belongs to the X-ray computed tomography facility of the Department of Development and Regeneration of the KU Leuven, financed by the Hercules Foundation (project AKUL/13/47). Immunofluorescence images were captured on a Zeiss LSM 880, Cell and Tissue Imaging Cluster, supported by Hercules AKUL/15/37_GOH1816N and FWO G.0929.15 to Pieter Vanden Berghe, University of Leuven.