Human lung fibroblast-to-myofibroblast transformation is not driven by an LDH5-dependent metabolic shift towards aerobic glycolysis.
Aerobic glycolysis
Fibroblast-to-myofibroblast transformation
Human lung fibroblasts
Idiopathic pulmonary fibrosis
Lactate dehydrogenase
Metabolic shift
TGF-β1
Journal
Respiratory research
ISSN: 1465-993X
Titre abrégé: Respir Res
Pays: England
ID NLM: 101090633
Informations de publication
Date de publication:
09 May 2019
09 May 2019
Historique:
received:
04
02
2019
accepted:
24
04
2019
entrez:
11
5
2019
pubmed:
11
5
2019
medline:
25
12
2019
Statut:
epublish
Résumé
Idiopathic pulmonary fibrosis (IPF) is a fatal respiratory disease characterized by aberrant fibroblast activation and progressive fibrotic remodelling of the lungs. Though the exact pathophysiological mechanisms of IPF remain unknown, TGF-β1 is thought to act as a main driver of the disease by mediating fibroblast-to-myofibroblast transformation (FMT). Recent reports have indicated that a metabolic shift towards aerobic glycolysis takes place during FMT and that metabolic shifts can directly influence aberrant cell function. This has led to the hypothesis that inhibition of lactate dehydrogenase 5 (LDH5), an enzyme responsible for converting pyruvate into lactate, could constitute a therapeutic concept for IPF. In this study, we investigated the potential link between aerobic glycolysis and FMT using a potent LDH5 inhibitor (Compound 408, Genentech). Seahorse analysis was performed to determine the effect of Compound 408 on TGF-β1-driven glycolysis in WI-38 fibroblasts. TGF-β1-mediated FMT was measured by quantifying α-smooth muscle actin (α-SMA) and fibronectin in primary human lung fibroblasts following treatment with Compound 408. Lactate and pyruvate levels in the cell culture supernatant were assessed by LC-MS/MS. In addition to pharmacological LDH5 inhibition, the effect of siRNA-mediated knockdown of LDHA and LDHB on FMT was examined. We show that treatment of lung fibroblasts with Compound 408 efficiently inhibits LDH5 and attenuates the TGF-β1-mediated metabolic shift towards aerobic glycolysis. Additionally, we demonstrate that LDH5 inhibition has no significant effect on TGF-β1-mediated FMT in primary human lung fibroblasts by analysing α-SMA fibre formation and fibronectin expression. Our data strongly suggest that while LDH5 inhibition can prevent metabolic shifts in fibroblasts, it has no influence on FMT and therefore glycolytic dysregulation is unlikely to be the sole driver of FMT.
Sections du résumé
BACKGROUND
BACKGROUND
Idiopathic pulmonary fibrosis (IPF) is a fatal respiratory disease characterized by aberrant fibroblast activation and progressive fibrotic remodelling of the lungs. Though the exact pathophysiological mechanisms of IPF remain unknown, TGF-β1 is thought to act as a main driver of the disease by mediating fibroblast-to-myofibroblast transformation (FMT). Recent reports have indicated that a metabolic shift towards aerobic glycolysis takes place during FMT and that metabolic shifts can directly influence aberrant cell function. This has led to the hypothesis that inhibition of lactate dehydrogenase 5 (LDH5), an enzyme responsible for converting pyruvate into lactate, could constitute a therapeutic concept for IPF.
METHODS
METHODS
In this study, we investigated the potential link between aerobic glycolysis and FMT using a potent LDH5 inhibitor (Compound 408, Genentech). Seahorse analysis was performed to determine the effect of Compound 408 on TGF-β1-driven glycolysis in WI-38 fibroblasts. TGF-β1-mediated FMT was measured by quantifying α-smooth muscle actin (α-SMA) and fibronectin in primary human lung fibroblasts following treatment with Compound 408. Lactate and pyruvate levels in the cell culture supernatant were assessed by LC-MS/MS. In addition to pharmacological LDH5 inhibition, the effect of siRNA-mediated knockdown of LDHA and LDHB on FMT was examined.
RESULTS
RESULTS
We show that treatment of lung fibroblasts with Compound 408 efficiently inhibits LDH5 and attenuates the TGF-β1-mediated metabolic shift towards aerobic glycolysis. Additionally, we demonstrate that LDH5 inhibition has no significant effect on TGF-β1-mediated FMT in primary human lung fibroblasts by analysing α-SMA fibre formation and fibronectin expression.
CONCLUSIONS
CONCLUSIONS
Our data strongly suggest that while LDH5 inhibition can prevent metabolic shifts in fibroblasts, it has no influence on FMT and therefore glycolytic dysregulation is unlikely to be the sole driver of FMT.
Identifiants
pubmed: 31072408
doi: 10.1186/s12931-019-1058-2
pii: 10.1186/s12931-019-1058-2
pmc: PMC6507142
doi:
Substances chimiques
Enzyme Inhibitors
0
Lactate Dehydrogenase 5
EC 1.1.1.27.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
87Subventions
Organisme : MRF
ID : MRF_MRF-091-0001-RG-GARNE
Pays : United Kingdom
Références
Life Sci. 2000 Oct 20;67(22):2663-71
pubmed: 11105982
Respir Res. 2002;3:3
pubmed: 11806838
Planta Med. 1992 Oct;58(5):454-8
pubmed: 17226502
Am J Pathol. 2007 Jun;170(6):1807-16
pubmed: 17525249
Chest. 2007 Oct;132(4):1311-21
pubmed: 17934117
Br J Pharmacol. 2009 Nov;158(5):1196-209
pubmed: 19785660
Cell Cycle. 2009 Dec;8(23):3984-4001
pubmed: 19923890
Hematology. 2010 Jun;15(3):144-50
pubmed: 20557672
Am J Respir Crit Care Med. 2011 Mar 15;183(6):788-824
pubmed: 21471066
Lancet. 2011 Dec 3;378(9807):1949-61
pubmed: 21719092
Cancer Res. 1990 Nov 1;50(21):6936-43
pubmed: 2208159
Am J Respir Crit Care Med. 2012 Oct 15;186(8):740-51
pubmed: 22923663
Eur Respir J. 2013 May;41(5):1207-18
pubmed: 23100500
Annu Rev Pathol. 2014;9:157-79
pubmed: 24050627
Future Med Chem. 2013 Oct;5(16):1967-91
pubmed: 24175747
Am J Respir Crit Care Med. 2014 May 15;189(10):1161-72
pubmed: 24641682
Mayo Clin Proc. 2014 Aug;89(8):1130-42
pubmed: 24867394
Eur Respir J. 2015 Sep;46(3):795-806
pubmed: 25976683
Am J Respir Crit Care Med. 2015 Dec 15;192(12):1462-74
pubmed: 26284610
J Biol Chem. 2015 Oct 16;290(42):25427-38
pubmed: 26318453
J Med Chem. 2016 Jan 28;59(2):487-96
pubmed: 26340601
Am J Physiol Lung Cell Mol Physiol. 2015 Dec 1;309(11):L1305-12
pubmed: 26408551
Toxicol Appl Pharmacol. 2016 Feb 1;292:56-64
pubmed: 26765310
World J Surg Oncol. 2016 Jan 20;14(1):15
pubmed: 26791262
Chest. 2016 Mar;149(3):756-66
pubmed: 26836914
J Proteome Res. 2016 May 6;15(5):1717-24
pubmed: 27052453
Future Med Chem. 2016 Apr;8(6):713-25
pubmed: 27054686
Drug Chem Toxicol. 2016 Oct;39(4):357-61
pubmed: 27071859
F1000Res. 2016 May 31;5:null
pubmed: 27303645
Nat Chem Biol. 2016 Oct;12(10):779-86
pubmed: 27479743
Sci Rep. 2017 Dec;7(1):149
pubmed: 28273952
Radiat Res. 2017 Jul;188(1):35-43
pubmed: 28463588
Br J Pharmacol. 2017 Nov;174(21):3848-3864
pubmed: 28810065
Sci Rep. 2017 Aug 16;7(1):8489
pubmed: 28814730
Lab Invest. 2017 Nov;97(11):1321-1331
pubmed: 28846077
BMJ Open Respir Res. 2017 Jun 5;4(1):e000183
pubmed: 28883924
Mol Med Rep. 2017 Dec;16(6):8335-8344
pubmed: 28983605
Am J Physiol Lung Cell Mol Physiol. 2018 Apr 1;314(4):L544-L554
pubmed: 29351437
Onco Targets Ther. 2018 Apr 27;11:2363-2373
pubmed: 29740212
PLoS One. 2018 May 24;13(5):e0197936
pubmed: 29795645
Cancer Res. 1984 Feb;44(2):768-71
pubmed: 6581864