Aortic carboxypeptidase-like protein (ACLP) enhances lung myofibroblast differentiation through transforming growth factor β receptor-dependent and -independent pathways

J Biol Chem. 2014 Jan 31;289(5):2526-36. doi: 10.1074/jbc.M113.502617. Epub 2013 Dec 16.

Abstract

Idiopathic pulmonary fibrosis (IPF) is a chronic and fatal lung disease characterized by the overgrowth, hardening, and scarring of lung tissue. The exact mechanisms of how IPF develops and progresses are unknown. IPF is characterized by extracellular matrix remodeling and accumulation of active TGFβ, which promotes collagen expression and the differentiation of smooth muscle α-actin (SMA)-positive myofibroblasts. Aortic carboxypeptidase-like protein (ACLP) is an extracellular matrix protein secreted by fibroblasts and myofibroblasts and is expressed in fibrotic human lung tissue and in mice with bleomycin-induced fibrosis. Importantly, ACLP knockout mice are significantly protected from bleomycin-induced fibrosis. The goal of this study was to identify the mechanisms of ACLP action on fibroblast differentiation. As primary lung fibroblasts differentiated into myofibroblasts, ACLP expression preceded SMA and collagen expression. Recombinant ACLP induced SMA and collagen expression in mouse and human lung fibroblasts. Knockdown of ACLP slowed the fibroblast-to-myofibroblast transition and partially reverted differentiated myofibroblasts by reducing SMA expression. We hypothesized that ACLP stimulates myofibroblast formation partly through activating TGFβ signaling. Treatment of fibroblasts with recombinant ACLP induced phosphorylation and nuclear translocation of Smad3. This phosphorylation and induction of SMA was dependent on TGFβ receptor binding and kinase activity. ACLP-induced collagen expression was independent of interaction with the TGFβ receptor. These findings indicate that ACLP stimulates the fibroblast-to-myofibroblast transition by promoting SMA expression via TGFβ signaling and promoting collagen expression through a TGFβ receptor-independent pathway.

Keywords: ACLP; Collagen; Myofibroblast; Pulmonary Fibrosis; SMAD Transcription Factor; Transforming Growth Factor β (TGFβ).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibiotics, Antineoplastic / toxicity
  • Bleomycin / toxicity
  • Carboxypeptidases / genetics
  • Carboxypeptidases / metabolism*
  • Cell Differentiation / physiology
  • Collagen / genetics
  • Collagen / metabolism
  • Disease Models, Animal
  • Fibroblasts / cytology*
  • Fibroblasts / metabolism
  • HEK293 Cells
  • Humans
  • Idiopathic Pulmonary Fibrosis / metabolism*
  • Idiopathic Pulmonary Fibrosis / pathology
  • Lung / cytology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mink
  • Primary Cell Culture
  • Receptors, Transforming Growth Factor beta / metabolism*
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Respiratory Mucosa / cytology
  • Respiratory Mucosa / metabolism
  • Signal Transduction / physiology*

Substances

  • AEBP1 protein, human
  • Aebp1 protein, mouse
  • Antibiotics, Antineoplastic
  • Receptors, Transforming Growth Factor beta
  • Repressor Proteins
  • Bleomycin
  • Collagen
  • Carboxypeptidases