Reduced FoxO3a expression causes low autophagy in idiopathic pulmonary fibrosis fibroblasts on collagen matrices.

Abstract

Idiopathic pulmonary fibrosis (IPF) is a chronic and fatal lung disease, and fibroblasts derived from patients with IPF are resistant to type I collagen matrix-induced cell death. The alteration of the PTEN-Akt axis permits IPF fibroblasts to maintain a pathological phenotype on collagen by suppressing autophagy. However, the precise underlying mechanism by which the Akt downstream molecule suppresses autophagic activity remains elusive. FoxO3a is a direct target of Akt and is implicated with the transcriptional activation of autophagy. Therefore, we investigated whether reduced FoxO3a expression causes abnormally low autophagy in IPF fibroblasts on collagen. We found that FoxO3a mRNA and protein levels are low in IPF fibroblasts, which subsequently suppresses the autophagosomal marker LC3B expression on collagen matrix. In contrast, the majority of control fibroblasts showed an increase in FoxO3a and LC3B expression at both the mRNA and protein levels. The luciferase assay confirmed that FoxO3a binds to the promoter region of LC3B and transcriptionally activates LC3B. The overexpression of wild-type FoxO3a increased LC3B mRNA and protein expression in IPF fibroblasts, whereas the dominant negative FoxO3a decreased the LC3B level in control fibroblasts. The inhibition of autophagic activity sensitized control fibroblasts to collagen matrix-induced cell death. In contrast, enhanced viability was found when autophagic function was inhibited in IPF fibroblasts. Our study showed that aberrantly low FoxO3a expression participates in reducing autophagic activity via transcriptional suppression of LC3B in IPF fibroblasts on collagen. This suggests that low autophagic activity by the alteration of FoxO3a may contribute to IPF progression.

• Authors: Jintaek Im, Polla Hergert, Richard Seonghun Nho
• Journal: American journal of physiology. Lung cellular and molecular physiology (Am J Physiol Lung Cell Mol Physiol) Vol. 309 Issue 6 Pg. L552-61 (Sep 15 2015) ISSN: 1522-1504 [Electronic] United States
• PMID: 26186945 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2015 the American Physiological Society.

Chemical References

• Culture Media
• FOXO3 protein, human
• Forkhead Box Protein O3
• Forkhead Transcription Factors
• MAP1LC3B protein, human
• Microtubule-Associated Proteins
• Collagen

Topics

• Autophagy
• Cells, Cultured
• Collagen (chemistry)
• Culture Media (chemistry)
• Disease Progression
• Fibroblasts (physiology)
• Forkhead Box Protein O3
• Forkhead Transcription Factors (genetics, metabolism)
• Gene Expression
• Gene Silencing
• Humans
• Idiopathic Pulmonary Fibrosis (metabolism, pathology)
• Microtubule-Associated Proteins (genetics, metabolism)