STAT3 regulates the onset of oxidant-induced senescence in lung fibroblasts

David W. Waters, Kaj E. C. Blokland, Prabuddha S. Pathinayake, Lan Wei, Michael Schuliga, Jade Jaffar, Glen P. Westall, Philip M. Hansbro, Cecilia M. Prele, Steven E. Mutsaers, Nathan W. Bartlett, Janette K. Burgess, Christopher L. Grainge, Darryl A. Knight

Research output: Contribution to journalArticle

5 Citations (Scopus)
309 Downloads (Pure)

Abstract

Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease of unknown cause with a median survival of only 3 years. We and others have shown that fibroblasts derived from IPF-lungs display characteristics of senescent cells and that dysregulated activation of the transcription factor signal transducer and activator of transcription 3 (STAT3) correlates with IPF progression. The question of whether STAT3 activation is involved in fibroblast senescence remains unanswered. We hypothesized that inhibiting STAT3 activation after oxidant-induced senescence would attenuate characteristics of the senescent phenotype. We aimed to characterize a model of oxidant induced senescence in human lung fibroblasts and to determine the effect of inhibiting STAT3 activity on the development of senescence. Exposing human lung fibroblasts to 150µM hydrogen peroxide (H2O2) resulted in increased senescence-associated-β-galactosidase (SA-β-Gal) content, expression of p21 and interleukin (IL)-6, all of which are features of senescence. The shift into senescence was accompanied by an increase of STAT3 translocation to the nucleus and mitochondria. Additionally, Seahorse analysis provided evidence of increased mitochondrial respiration characterized by increased basal respiration, proton leak and an associated increase in superoxide (O2-) production in senescent fibroblasts. Targeting STAT3 activity using the small molecule inhibitor STA-21 attenuated IL-6 production, reduced p21 levels, decreased SA-β-Gal accumulation and restored normal mitochondrial function. The results of this study illustrate that stress-induced senescence in lung fibroblasts involves the activation of STAT3 which can be pharmacologically modulated.

Original languageEnglish
JournalAmerican Journal of Respiratory Cell and Molecular Biology
DOIs
Publication statusPublished - Jul 2019

Fingerprint

STAT3 Transcription Factor
Fibroblasts
Oxidants
Lung
Chemical activation
Idiopathic Pulmonary Fibrosis
Galactosidases
Interleukin-6
Transcriptional Activation
Respiration
Pulmonary diseases
Mitochondria
Smegmamorpha
Superoxides
Hydrogen Peroxide
Protons
Transcription Factors
Display devices
Lung Diseases
Chronic Disease

Cite this

Waters, D. W., Blokland, K. E. C., Pathinayake, P. S., Wei, L., Schuliga, M., Jaffar, J., ... Knight, D. A. (2019). STAT3 regulates the onset of oxidant-induced senescence in lung fibroblasts. American Journal of Respiratory Cell and Molecular Biology. https://doi.org/10.1165/rcmb.2018-0328OC
Waters, David W. ; Blokland, Kaj E. C. ; Pathinayake, Prabuddha S. ; Wei, Lan ; Schuliga, Michael ; Jaffar, Jade ; Westall, Glen P. ; Hansbro, Philip M. ; Prele, Cecilia M. ; Mutsaers, Steven E. ; Bartlett, Nathan W. ; Burgess, Janette K. ; Grainge, Christopher L. ; Knight, Darryl A. / STAT3 regulates the onset of oxidant-induced senescence in lung fibroblasts. In: American Journal of Respiratory Cell and Molecular Biology. 2019.
@article{4e31294639a1439a9ceebfc067715cbb,
title = "STAT3 regulates the onset of oxidant-induced senescence in lung fibroblasts",
abstract = "Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease of unknown cause with a median survival of only 3 years. We and others have shown that fibroblasts derived from IPF-lungs display characteristics of senescent cells and that dysregulated activation of the transcription factor signal transducer and activator of transcription 3 (STAT3) correlates with IPF progression. The question of whether STAT3 activation is involved in fibroblast senescence remains unanswered. We hypothesized that inhibiting STAT3 activation after oxidant-induced senescence would attenuate characteristics of the senescent phenotype. We aimed to characterize a model of oxidant induced senescence in human lung fibroblasts and to determine the effect of inhibiting STAT3 activity on the development of senescence. Exposing human lung fibroblasts to 150µM hydrogen peroxide (H2O2) resulted in increased senescence-associated-β-galactosidase (SA-β-Gal) content, expression of p21 and interleukin (IL)-6, all of which are features of senescence. The shift into senescence was accompanied by an increase of STAT3 translocation to the nucleus and mitochondria. Additionally, Seahorse analysis provided evidence of increased mitochondrial respiration characterized by increased basal respiration, proton leak and an associated increase in superoxide (O2-) production in senescent fibroblasts. Targeting STAT3 activity using the small molecule inhibitor STA-21 attenuated IL-6 production, reduced p21 levels, decreased SA-β-Gal accumulation and restored normal mitochondrial function. The results of this study illustrate that stress-induced senescence in lung fibroblasts involves the activation of STAT3 which can be pharmacologically modulated.",
author = "Waters, {David W.} and Blokland, {Kaj E. C.} and Pathinayake, {Prabuddha S.} and Lan Wei and Michael Schuliga and Jade Jaffar and Westall, {Glen P.} and Hansbro, {Philip M.} and Prele, {Cecilia M.} and Mutsaers, {Steven E.} and Bartlett, {Nathan W.} and Burgess, {Janette K.} and Grainge, {Christopher L.} and Knight, {Darryl A.}",
year = "2019",
month = "7",
doi = "10.1165/rcmb.2018-0328OC",
language = "English",
journal = "American Journal of Repiratory Cell and Molecular Biology",
issn = "1044-1549",
publisher = "AMER THORACIC SOC",

}

Waters, DW, Blokland, KEC, Pathinayake, PS, Wei, L, Schuliga, M, Jaffar, J, Westall, GP, Hansbro, PM, Prele, CM, Mutsaers, SE, Bartlett, NW, Burgess, JK, Grainge, CL & Knight, DA 2019, 'STAT3 regulates the onset of oxidant-induced senescence in lung fibroblasts' American Journal of Respiratory Cell and Molecular Biology. https://doi.org/10.1165/rcmb.2018-0328OC

STAT3 regulates the onset of oxidant-induced senescence in lung fibroblasts. / Waters, David W.; Blokland, Kaj E. C.; Pathinayake, Prabuddha S.; Wei, Lan; Schuliga, Michael; Jaffar, Jade; Westall, Glen P.; Hansbro, Philip M.; Prele, Cecilia M.; Mutsaers, Steven E.; Bartlett, Nathan W.; Burgess, Janette K.; Grainge, Christopher L.; Knight, Darryl A.

In: American Journal of Respiratory Cell and Molecular Biology, 07.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - STAT3 regulates the onset of oxidant-induced senescence in lung fibroblasts

AU - Waters, David W.

AU - Blokland, Kaj E. C.

AU - Pathinayake, Prabuddha S.

AU - Wei, Lan

AU - Schuliga, Michael

AU - Jaffar, Jade

AU - Westall, Glen P.

AU - Hansbro, Philip M.

AU - Prele, Cecilia M.

AU - Mutsaers, Steven E.

AU - Bartlett, Nathan W.

AU - Burgess, Janette K.

AU - Grainge, Christopher L.

AU - Knight, Darryl A.

PY - 2019/7

Y1 - 2019/7

N2 - Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease of unknown cause with a median survival of only 3 years. We and others have shown that fibroblasts derived from IPF-lungs display characteristics of senescent cells and that dysregulated activation of the transcription factor signal transducer and activator of transcription 3 (STAT3) correlates with IPF progression. The question of whether STAT3 activation is involved in fibroblast senescence remains unanswered. We hypothesized that inhibiting STAT3 activation after oxidant-induced senescence would attenuate characteristics of the senescent phenotype. We aimed to characterize a model of oxidant induced senescence in human lung fibroblasts and to determine the effect of inhibiting STAT3 activity on the development of senescence. Exposing human lung fibroblasts to 150µM hydrogen peroxide (H2O2) resulted in increased senescence-associated-β-galactosidase (SA-β-Gal) content, expression of p21 and interleukin (IL)-6, all of which are features of senescence. The shift into senescence was accompanied by an increase of STAT3 translocation to the nucleus and mitochondria. Additionally, Seahorse analysis provided evidence of increased mitochondrial respiration characterized by increased basal respiration, proton leak and an associated increase in superoxide (O2-) production in senescent fibroblasts. Targeting STAT3 activity using the small molecule inhibitor STA-21 attenuated IL-6 production, reduced p21 levels, decreased SA-β-Gal accumulation and restored normal mitochondrial function. The results of this study illustrate that stress-induced senescence in lung fibroblasts involves the activation of STAT3 which can be pharmacologically modulated.

AB - Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease of unknown cause with a median survival of only 3 years. We and others have shown that fibroblasts derived from IPF-lungs display characteristics of senescent cells and that dysregulated activation of the transcription factor signal transducer and activator of transcription 3 (STAT3) correlates with IPF progression. The question of whether STAT3 activation is involved in fibroblast senescence remains unanswered. We hypothesized that inhibiting STAT3 activation after oxidant-induced senescence would attenuate characteristics of the senescent phenotype. We aimed to characterize a model of oxidant induced senescence in human lung fibroblasts and to determine the effect of inhibiting STAT3 activity on the development of senescence. Exposing human lung fibroblasts to 150µM hydrogen peroxide (H2O2) resulted in increased senescence-associated-β-galactosidase (SA-β-Gal) content, expression of p21 and interleukin (IL)-6, all of which are features of senescence. The shift into senescence was accompanied by an increase of STAT3 translocation to the nucleus and mitochondria. Additionally, Seahorse analysis provided evidence of increased mitochondrial respiration characterized by increased basal respiration, proton leak and an associated increase in superoxide (O2-) production in senescent fibroblasts. Targeting STAT3 activity using the small molecule inhibitor STA-21 attenuated IL-6 production, reduced p21 levels, decreased SA-β-Gal accumulation and restored normal mitochondrial function. The results of this study illustrate that stress-induced senescence in lung fibroblasts involves the activation of STAT3 which can be pharmacologically modulated.

U2 - 10.1165/rcmb.2018-0328OC

DO - 10.1165/rcmb.2018-0328OC

M3 - Article

JO - American Journal of Repiratory Cell and Molecular Biology

JF - American Journal of Repiratory Cell and Molecular Biology

SN - 1044-1549

ER -