Expression of a human surfactant protein C mutation associated with interstitial lung disease disrupts lung development in transgenic mice

J Biol Chem. 2003 Dec 26;278(52):52739-46. doi: 10.1074/jbc.M309599200. Epub 2003 Oct 2.

Abstract

Surfactant Protein C (SP-C) is a secreted transmembrane protein that is exclusively expressed by alveolar type II epithelial cells of the lung. SP-C associates with surfactant lipids to reduce surface tension within the alveolus, maintaining lung volume at end expiration. Mutations in the gene encoding SP-C (SFTPC) have recently been linked to chronic lung disease in children and adults. The goal of this study was to determine whether a disease-linked mutation in SFTPC causes lung disease in transgenic mice. The SFTPC mutation, designated g.1728 G --> A, results in the deletion of exon4, generating a truncated form of SP-C (SP-C(Deltaexon4)). cDNA encoding SP-C(Deltaexon4) was constitutively expressed in type II epithelial cells of transgenic mice. Viable F0 transgene-positive mice were not generated after two separate rounds of pronuclear injections. Histological analysis of lung tissue harvested from embryonic day 17.5 F0 transgene-positive fetuses revealed that SP-C(Deltaexon4) caused a dose-dependent disruption in branching morphogenesis of the lung associated with epithelial cell cytotoxicity. Transient expression of SP-C(Deltaexon4) in isolated type II epithelial cells or HEK293 cells resulted in incomplete processing of the mutant proprotein, a dose-dependent increase in BiP transcription, trapping of the proprotein in the endoplasmic reticulum, and rapid degradation via a proteasome-dependent pathway. Taken together, these data suggest that the g.1728 G --> A mutation causes misfolding of the SP-C proprotein with subsequent induction of the unfolded protein response and endoplasmic reticulum-associated degradation pathways ultimately resulting in disrupted lung morphogenesis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Western
  • Carrier Proteins / genetics
  • Cell Line
  • DNA / metabolism
  • DNA, Complementary / metabolism
  • Dose-Response Relationship, Drug
  • Endoplasmic Reticulum / metabolism
  • Endoplasmic Reticulum Chaperone BiP
  • Exons
  • Gene Deletion
  • Heat-Shock Proteins*
  • Humans
  • Intercellular Signaling Peptides and Proteins
  • Lipid Metabolism
  • Luciferases / metabolism
  • Lung / growth & development*
  • Lung / pathology
  • Lung Diseases / genetics*
  • Mice
  • Mice, Transgenic
  • Microscopy, Fluorescence
  • Models, Genetic
  • Molecular Chaperones / genetics
  • Mutation*
  • Peptides / genetics*
  • Peptides / metabolism*
  • Phenotype
  • Protein Binding
  • Protein Folding
  • Protein Transport
  • Pulmonary Surfactant-Associated Protein C
  • Time Factors
  • Transcription, Genetic
  • Transfection
  • Transgenes

Substances

  • Carrier Proteins
  • DNA, Complementary
  • Endoplasmic Reticulum Chaperone BiP
  • Heat-Shock Proteins
  • Intercellular Signaling Peptides and Proteins
  • Molecular Chaperones
  • Peptides
  • Pulmonary Surfactant-Associated Protein C
  • SFTPC protein, human
  • Sftpc protein, mouse
  • DNA
  • Luciferases