RESEARCH ARTICLE

Loss of IκB kinase β promotes myofibroblast transformation and senescence through activation of the ROS-TGFβ autocrine loop

  • Liang Chen 1 ,
  • Zhimin Peng 1 ,
  • Qinghang Meng 1 ,
  • Maureen Mongan 1 ,
  • Jingcai Wang 1 ,
  • Maureen Sartor 1 ,
  • Jing Chen 1 ,
  • Liang Niu 1 ,
  • Mario Medvedovic 1 ,
  • Winston Kao 2 ,
  • Ying Xia , 1,2
Expand
  • 1. Department of Environmental Health and Center of Environmental Genetics, University of Cincinnati Medical Center, Cincinnati, OH 45267, USA
  • 2. Department of Ophthalmology, University of Cincinnati Medical Center, Cincinnati, OH 45267, USA

Received date: 16 Nov 2015

Accepted date: 01 Dec 2015

Published date: 27 May 2016

Copyright

2016 The Author(s) 2016. This article is published with open access at Springerlink.com and journal.hep.com.cn

Abstract

Using forward and reverse genetics and global gene expression analyses, we explored the crosstalk between the IκB kinase β (IKKβ) and the transforming growth factor β (TGFβ) signaling pathways. We show that in vitro ablation of Ikkβin fibroblasts led to progressive ROS accumulation and TGFβ activation, and ultimately accelerated cell migration, fibroblast-myofibroblast transformation and senescence. Mechanistically, the basal IKKβ activity was required for anti-oxidant gene expression and redox homeostasis. Lacking this activity, IKKβ-null cells showed ROS accumulation and activation of stress-sensitive transcription factor AP-1/c-Jun. AP-1/c-Jun activation led to up-regulation of the Tgfβ2promoter, which in turn further potentiated intracellular ROS through the induction of NADPH oxidase (NOX). These data suggest that by blocking the autocrine amplification of a ROS-TGFβ loop IKKβ plays a crucial role in the prevention of fibroblast-myofibroblast transformation and senescence.

Cite this article

Liang Chen , Zhimin Peng , Qinghang Meng , Maureen Mongan , Jingcai Wang , Maureen Sartor , Jing Chen , Liang Niu , Mario Medvedovic , Winston Kao , Ying Xia . Loss of IκB kinase β promotes myofibroblast transformation and senescence through activation of the ROS-TGFβ autocrine loop[J]. Protein & Cell, 2016 , 7(5) : 338 -350 . DOI: 10.1007/s13238-015-0241-6

1
Aaronson SA, Todaro GJ (1968) Development of 3T3-like lines from Balb-c mouse embryo cultures: transformation susceptibility to SV40. J Cell Physiol 72:141–148

DOI

2
Anderson ME (1998) Glutathione: an overview of biosynthesis and modulation. Chem Biol Interact111–112:1–14

DOI

3
Armstrong JS, Steinauer KK, Hornung B, Irish JM, Lecane P, Birrell GW, Peehl DM, Knox SJ (2002) Role of glutathione depletion and reactive oxygen species generation in apoptotic signaling in a human B lymphoma cell line. Cell Death Differ 9:252–263

DOI

4
Bacher S, Schmitz ML (2004) The NF-kappaB pathway as a potential target for autoimmune disease therapy. Curr Pharm Des 10:2827–2837

DOI

5
Balaban RS, Nemoto S, Finkel T (2005) Mitochondria, oxidants, and aging. Cell 120:483–495

DOI

6
Baldwin AS Jr (2001) Series introduction: the transcription factor NFkappaB and human disease. J Clin Investig 107:3–6

DOI

7
Bataller R (2003) NADPH oxidase signal transduces angiotensin II in hepatic stellate cells and is critical in hepatic fibrosis. J Clin Investig 112:1383–1394

DOI

8
Bedard K, Krause KH (2007) The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology. Physiol Rev 87:245–313

DOI

9
Bitzer M, von Gersdorff G, Liang D, Dominguez-Rosales A, Beg AA, RojkindM Bottinger EP, (2000) Amechanismof suppression of TGFbeta/SMAD signaling by NF-kappa B/RelA. Genes Dev 14:187–197

10
Bondi CD, Manickam N, Lee DY, Block K, Gorin Y, Abboud HE, Barnes JL (2010) NAD(P)H oxidase mediates TGF-beta1-induced activation of kidney myofibroblasts. J Am Soc Nephrol 21:93–102

DOI

11
Callicott RJ, Womack JE (2006) Real-time PCR assay for measurement of mouse telomeres. Comp Med 56:17–22

12
Chariot A (2009) The NF-kappaB-independent functions of IKK subunits in immunity and cancer. Trends Cell Biol 19:404–413

DOI

13
Chen F, Castranova V, Li Z, Karin M, Shi X (2003) Inhibitor of nuclear factor kappaB kinase deficiency enhances oxidative stress and prolongs c-Jun NH2-terminal kinase activation induced by arsenic. Cancer Res 63:7689–7693

14
Chen F, Lu Y, Castranova V, Li Z, Karin M (2006) Loss of Ikkbeta promotes migration and proliferation of mouse embryo fibroblast cells. J Biol Chem 281:37142–37149

DOI

15
Chen Y, Johansson E, Fan Y, Shertzer HG, Vasiliou V, Nebert DW, Dalton TP (2009) Early onset senescence occurs when fibroblasts lack the glutamate–cysteine ligase modifier subunit. Free Radic Biol Med 47:410–418

DOI

16
Courtois G, Gilmore TD (2006) Mutations in the NF-kappaB signaling pathway: implications for human disease. Oncogene 25:6831–6843

DOI

17
Datto MB, Frederick JP, Pan L, Borton AJ, Zhuang Y, Wang XF (1999) Targeted disruption of Smad3 reveals an essential role in transforming growth factor beta-mediated signal transduction. Mol Cell Biol 19:2495–2504

DOI

18
Debacq-Chainiaux F, Pascal T, Boilan E, Bastin C, Bauwens E, Toussaint O (2008) Screening of senescence-associated genes with specific DNA array reveals the role of IGFBP-3 in premature senescence of human diploid fibroblasts. Free Radic Biol Med 44:1817–1832

DOI

19
Dennler S, Prunier C, Ferrand N, Gauthier JM, Atfi A (2000) c-Jun inhibits transforming growth factor beta-mediated transcription by repressing Smad3 transcriptional activity. J Biol Chem 275:28858–28865

DOI

20
DiDonato JA, Mercurio F, Karin M (2012) NF-kappaB and the link between inflammation and cancer. Immunol Rev 246:379–400

DOI

21
Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereira-Smith O (1995) A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA 92:9363–9367

DOI

22
Dumont P, Burton M, Chen QM, Gonos ES, Frippiat C, Mazarati JB, Eliaers F, Remacle J, Toussaint O (2000) Induction of replicative senescence biomarkers by sublethal oxidative stresses in normal human fibroblast. Free Radic Biol Med 28:361–373

DOI

23
Fleckenstein K, Zgonjanin L, Chen L, Rabbani Z, Jackson IL, Thrasher B, Kirkpatrick J, Foster WM, Vujaskovic Z (2007) Temporal onset of hypoxia and oxidative stress after pulmonary irradiation. Int J Radiat Oncol Biol Phys 68:196–204

DOI

24
Freudenberg JM, Joshi VK, Hu Z, Medvedovic M (2009) CLEAN: clustering enrichment analysis. BMC Bioinform 10:234

DOI

25
Freudlsperger C, Bian Y, Contag WS, Burnett J, Coupar J, Yang X, Chen Z, Van WC (2013) TGF-beta and NF-kappaB signal pathway cross-talk is mediated through TAK1 and SMAD7 in a subset of head and neck cancers. Oncogene 32:1549–1559

DOI

26
Geh E, Meng Q, Mongan M, Wang J, Takatori A, Zheng Y, Puga A, Lang RA, Xia Y (2011) Mitogen-activated protein kinase kinase kinase 1 (MAP3K1) integrates developmental signals for eyelid closure. Proc Natl Acad Sci USA 108:17349–17354

DOI

27
Gerondakis S, Grumont R, Gugasyan R, Wong L, Isomura I, Ho W, Banerjee A (2006) Unravelling the complexities of the NF-kappaB signalling pathway using mouse knockout and transgenic models. Oncogene 25:6781–6799

DOI

28
Gingery A, Bradley EW, Pederson L, Ruan M, Horwood NJ, Oursler MJ (2008) TGF-beta coordinately activates TAK1/MEK/AKT/NFkB and SMAD pathways to promote osteoclast survival. Exp Cell Res 314:2725–2738

DOI

29
Giorgio M, Trinei M, Migliaccio E, Pelicci PG (2007) Hydrogen peroxide: a metabolic by-product or a common mediator of ageing signals? Nat Rev Mol Cell Biol 8:722–728

30
Hecker L, Vittal R, Jones T, Jagirdar R, Luckhardt TR, Horowitz JC, Pennathur S, Martinez FJ, Thannickal VJ (2009) NADPH oxidase-4 mediates myofibroblast activation and fibrogenic responses to lung injury. Nat Med 15:1077–1081

DOI

31
Irizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, Scherf U, Speed TP (2003) Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4:249–264

DOI

32
Karin M (2006) Nuclear factor-kappaB in cancer development and progression. Nature 441:431–436

DOI

33
Karin M (2008) The IkappaB kinase: a bridge between inflammation and cancer. Cell Res 18:334–342

DOI

34
Kim HJ, Hawke N, Baldwin AS (2006) NF-kappaB and IKK as therapeutic targets in cancer 3. Cell Death Differ 13:738–747

DOI

35
Lavon I (2000) High susceptibility to bacterial infection, but no liver dysfunction, in mice compromised for hepatocyte NFkappaB activation. Nat Med 6:573–577

DOI

36
Li X, Massa PE, Hanidu A, Peet GW, Aro P, Savitt A, Mische S, Li J, Marcu KB (2002) IKKalpha, IKKbeta, and NEMO/IKKgamma are each required for the NF-kappa B-mediated inflammatory response program. J Biol Chem 277:45129–45140

DOI

37
Luo JL, Kamata H, Karin M (2005) IKK/NF-kappaB signaling: balancing life and death–a new approach to cancer therapy. J Clin Investig 115:2625–2632

DOI

38
Maeda S, Kamata H, Luo JL, Leffert H, Karin M (2005) IKKbeta couples hepatocyte death to cytokine-driven compensatory proliferation that promotes chemical hepatocarcinogenesis 1. Cell 121:977–990

DOI

39
May MJ, Madge LA (2007) Caspase inhibition sensitizes inhibitor of NF-kappaB kinase beta-deficient fibroblasts to caspase-independent cell death via the generation of reactive oxygen species. J Biol Chem 282:16105–16116

DOI

40
McIntyre KW, Shuster DJ, Gillooly KM, Dambach DM, Pattoli MA, Lu P, Zhou XD, Qiu Y, Zusi FC, Burke JR (2003) A highly selective inhibitor of I kappa B kinase, BMS-345541, blocks both joint inflammation and destruction in collagen-induced arthritis in mice. Arthritis Rheum 48:2652–2659

DOI

41
Medvedovic M (2009) Influence of fatty acid diets on gene expression in rat mammary epithelial cells. Physiol Genom 38 (1):80–88

42
Nagarajan RP, Chen F, Li W, Vig E, Harrington MA, Nakshatri H, Chen Y (2000) Repression of transforming-growth-factor-betamediated transcription by nuclear factor kappaB. Biochem J 348( Pt 3):591–596

43
Nelson G, Wordsworth J, Wang C, Jurk D, Lawless C, Martin-Ruiz C, von Zglinicki T (2012) A senescent cell bystander effect: senescence-induced senescence. Aging Cell 11:345–349

DOI

44
O’Dea EL, Barken D, Peralta RQ, Tran KT, Werner SL, Kearns JD, Levchenko A, Hoffmann A (2007) A homeostatic model of IkappaB metabolism to control constitutive NF-kappaB activity. Mol Syst Biol 3:111

45
Pasparakis M (2009) Regulation of tissue homeostasis by NFkappaB signalling: implications for inflammatory diseases. Nat Rev Immunol 9:778–788

DOI

46
Passos JFl (2010) Feedback between p21 and reactive oxygen production is necessary for cell senescence. Mol Syst Biol 6:347

47
Peng Z, Peng L, Fan Y, Zandi E, Shertzer HG, Xia Y (2007) A critical role for IkappaB kinase beta in metallothionein-1 expression and protection against arsenic toxicity. J Biol Chem 282:21487–21496

DOI

48
Peng Z, Geh E, Chen L, Meng Q, Fan Y, Sartor M, Shertzer HG, Liu ZG, Puga A, Xia Y (2010) IkB kinase b regulates redox homeostasis by controlling the constitutive levels of glutathione. Mol Pharmacol 77(5):784–792

49
Perkins ND (2007) Integrating cell-signalling pathways with NFkappaB and IKK function. Nat Rev Mol Cell Biol 8:49–62

DOI

50
Phan SH (2002) The myofibroblast in pulmonary fibrosis. Chest 122:286S–289S

51
Polzer K, Baeten D, Soleiman A, Distler J, Gerlag DM, Tak PP, Schett G, Zwerina J (2008) Tumour necrosis factor blockade increases lymphangiogenesis in murine and human arthritic joints. Ann Rheum Dis 67:1610–1616

DOI

52
Roy S, Dontamalla SK, Mondru AK, Sannigrahi S, Veerareddy PR (2011) Downregulation of apoptosis and modulation of TGFbeta1 by sodium selenate prevents streptozotocin-induced diabetic rat renal impairment. Biol Trace Elem Res 139:55–71

DOI

53
Ruocco MG, Maeda S, Park JM, Lawrence T, Hsu LC, Cao Y, Schett G, Wagner EF, Karin M (2005) IkB kinase IKKb, but not IKKa, is a critical mediator of osteoclast survival and is required for inflammation-induced bone loss. J Exp Med 201:1677–1687

DOI

54
Sakon S (2003) NF-kappaB inhibits TNF-induced accumulation of ROS that mediate prolonged MAPK activation and necrotic cell death. EMBO J 22:3898–3909

DOI

55
Sartor MA, Tomlinson CR, Wesselkamper SC, Sivaganesan S, Leikauf GD, Medvedovic M (2006) Intensity-based hierarchical Bayes method improves testing for differentially expressed genes in microarray experiments. BMC Bioinform 7:538

DOI

56
Sartor MA, Leikauf GD, Medvedovic M (2009) LRpath: a logistic regression approach for identifying enriched biological groups in gene expression data. Bioinformatics 25:211–217

DOI

57
Schnekenburger M, Peng L, Puga A (2007) HDAC1 bound to the Cyp1a1 promoter blocks histone acetylation associated with Ah receptor-mediated trans-activation. Biochim Biophys Acta 1769:569–578

DOI

58
Sen CK, Roy S (2010) Oxygenation state as a driver of myofibroblast differentiation and wound contraction: hypoxia impairs wound closure. J Investig Dermatol 130:2701–2703

DOI

59
Tanaka M, Fuentes ME, Yamaguchi K, Durnin MH, Dalrymple SA, Hardy KL, Goeddel DV (1999) Embryonic lethality, liver degeneration, and impaired NF-kappa B activation in IKK-betadeficient mice. Immunity 10:421–429

DOI

60
Tojima Y (2000) NAK is an IkappaB kinase-activating kinase. Nature 404:778–782

DOI

61
Vallabhapurapu S, Karin M (2009) Regulation and function of NFkappaB transcription factors in the immune system. Annu Rev Immunol 27:693–733

DOI

62
Verrecchia F, Vindevoghel L, Lechleider RJ, Uitto J, Roberts AB, Mauviel A (2001) Smad3/AP-1 interactions control transcriptional responses to TGF-beta in a promoter-specific manner. Oncogene 20:3332–3340

DOI

63
Weyemi U (2011) ROS-generating NADPH oxidase NOX4 is a critical mediator in oncogenic H-Ras-induced DNA damage and subsequent senescence. Oncogene 31(9):1117–1129

64
Zhang L, Wang W, Hayashi Y, Jester JV, Birk DE, Gao M, Liu CY, Kao WW, Karin M, Xia Y (2003) A role for MEK kinase 1 in TGFbeta/activin-induced epithelium movement and embryonic eyelid closure. EMBO J 22:4443–4454

DOI

Outlines

/