TY - JOUR
T1 - Profibrotic epithelial TGF-β1 signaling involves NOX4-mitochondria cross talk and redox-mediated activation of the tyrosine kinase FYN
AU - Veith, Carmen
AU - Hristova, Milena
AU - Danyal, Karamatullah
AU - Habibovic, Aida
AU - Dustin, Christopher M.
AU - McDonough, John E.
AU - Vanaudenaerde, Bart M.
AU - Kreuter, Michael
AU - Schneider, Marc A.
AU - Kahn, Nicolas
AU - van Schooten, Frederik J.
AU - Boots, Agnes W.
AU - van der Vliet, Albert
N1 - Funding Information:
We thank Nicole Bishop for assistance with confocal microscopy imaging, which was performed on a Zeiss 510 META laser scanning confocal microscope supported by National Institutes of Health Award 1S10RR019246 from the National Center for Research Resources.
Funding Information:
This work was supported by National Institutes of Health Grants R01 HL085646, R01 HL138708, and R21 AG055325 (to A.vdV.). C.V. was supported by the Nutrim Graduate Programme from Maastricht University. This study was also supported by Lung Foundation Netherlands Grant 9.2.17.214FE (to C.V.), and the German Center for Lung Research Grant 82DZL00402. C.M.D. was supported by an F31 Predoctoral Fellowship from NIH (HL142221).
Publisher Copyright:
© 2021 American Physiological Society. All rights reserved.
PY - 2021/3
Y1 - 2021/3
N2 - Idiopathic pulmonary fibrosis (IPF) is characterized by a disturbed redox balance and increased production of reactive oxygen species (ROS), which is believed to contribute to epithelial injury and fibrotic lung scarring. The main pulmonary sources of ROS include mitochondria and NADPH oxidases (NOXs), of which the NOX4 isoform has been implicated in IPF. Non-receptor SRC tyrosine kinases (SFK) are important for cellular homeostasis and are often dysregulated in lung diseases. SFK activation by the profibrotic transforming growth factor-beta (TGF-beta) is thought to contribute to pulmonary fibrosis, but the relevant SFK isoform and its relationship to NOX4 and/or mitochondrial ROS in the context of profibrotic TGF-beta signaling is not known. Here, we demonstrate that TGF-beta 1 can rapidly activate the SRC kinase FYN in human bronchial epithelial cells, which subsequently induces mitochondrial ROS (mtROS) production, genetic damage shown by the DNA damage marker 7H2AX, and increased expression of profibrotic genes. Moreover, TGF-beta 1-induced activation of FYN involves initial activation of NOX4 and direct cysteine oxidation of FYN, and both FYN and mtROS contribute to TGF-beta 1-induced induction of NOX4. NOX4 expression in lung tissues of IPF patients is positively correlated with disease severity, although FYN expression is down-regulated in IPF and does not correlate with disease severity. Collectively, our findings highlight a critical role for FYN in TGF-beta 1-induced mtROS production, DNA damage response, and induction of profibrotic genes in bronchial epithelial cells, and suggest that altered expression and activation of NOX4 and FYN may contribute to the pathogenesis of pulmonary fibrosis.
AB - Idiopathic pulmonary fibrosis (IPF) is characterized by a disturbed redox balance and increased production of reactive oxygen species (ROS), which is believed to contribute to epithelial injury and fibrotic lung scarring. The main pulmonary sources of ROS include mitochondria and NADPH oxidases (NOXs), of which the NOX4 isoform has been implicated in IPF. Non-receptor SRC tyrosine kinases (SFK) are important for cellular homeostasis and are often dysregulated in lung diseases. SFK activation by the profibrotic transforming growth factor-beta (TGF-beta) is thought to contribute to pulmonary fibrosis, but the relevant SFK isoform and its relationship to NOX4 and/or mitochondrial ROS in the context of profibrotic TGF-beta signaling is not known. Here, we demonstrate that TGF-beta 1 can rapidly activate the SRC kinase FYN in human bronchial epithelial cells, which subsequently induces mitochondrial ROS (mtROS) production, genetic damage shown by the DNA damage marker 7H2AX, and increased expression of profibrotic genes. Moreover, TGF-beta 1-induced activation of FYN involves initial activation of NOX4 and direct cysteine oxidation of FYN, and both FYN and mtROS contribute to TGF-beta 1-induced induction of NOX4. NOX4 expression in lung tissues of IPF patients is positively correlated with disease severity, although FYN expression is down-regulated in IPF and does not correlate with disease severity. Collectively, our findings highlight a critical role for FYN in TGF-beta 1-induced mtROS production, DNA damage response, and induction of profibrotic genes in bronchial epithelial cells, and suggest that altered expression and activation of NOX4 and FYN may contribute to the pathogenesis of pulmonary fibrosis.
KW - idiopathic pulmonary fibrosis
KW - mitochondria
KW - NOX4
KW - redox signaling
KW - SRC
KW - IDIOPATHIC PULMONARY-FIBROSIS
KW - TGF-BETA
KW - OXIDATIVE STRESS
KW - NADPH-OXIDASE
KW - LUNG FIBROSIS
KW - MYOFIBROBLAST DIFFERENTIATION
KW - MITOCHONDRIAL-DNA
KW - SRC KINASE
KW - MIGRATION
U2 - 10.1152/ajplung.00444.2019
DO - 10.1152/ajplung.00444.2019
M3 - Article
C2 - 33325804
SN - 1040-0605
VL - 320
SP - L356-L367
JO - American Journal of Physiology-Lung Cellular and Molecular Physiology
JF - American Journal of Physiology-Lung Cellular and Molecular Physiology
IS - 3
ER -