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Open Access 01-12-2016 | Research article

Eosinophils enhance WNT-5a and TGF-β₁ genes expression in airway smooth muscle cells and promote their proliferation by increased extracellular matrix proteins production in asthma

Authors: Andrius Januskevicius, Simona Vaitkiene, Reinoud Gosens, Ieva Janulaityte, Deimante Hoppenot, Raimundas Sakalauskas, Kestutis Malakauskas

Published in: BMC Pulmonary Medicine | Issue 1/2016

Abstract

Background

Recent studies have suggested that eosinophils may have a direct effect on airway smooth muscle cells (ASMC), causing their proliferation in patients with asthma, but the precise mechanism of the interaction between these cells remains unknown. We propose that changes in Wnt signaling activity and extracellular matrix (ECM) production may help explain these findings. Therefore, the aim of this study was to investigate the effect of eosinophils from asthmatic and non-asthmatic subjects on Wnt-5a, transforming growth factor β₁ (TGF-β₁), and ECM protein (fibronectin and collagen) gene expression and ASMC proliferation.

Methods

A total of 18 subjects were involved in the study: 8 steroid-free asthma patients and 10 healthy subjects. Peripheral blood eosinophils were isolated using centrifugation and magnetic separation. An individual co-culture of eosinophils with human ASMC was prepared for each study subject. Adhesion of eosinophils to ASMC (evaluated by assaying eosinophil peroxidase activity) was determined following various incubation periods (30, 45, 60, 120, and 240 min). The expression of Wnt-5a, TGF-β₁, and ECM protein genes in ASMC was measured using quantitative real-time polymerase chain reaction (PCR) after 24 h of co-culture. Proliferation of ASMC was measured using the Alamar blue method after 48 h and 72 h of co-culture with eosinophils.

Results

Eosinophils from asthmatic subjects demonstrated increased adhesion to ASMC compared with eosinophils from healthy subjects (p < 0.05) in vitro. The expression of Wnt-5a, TGF-β₁, collagen, and fibronectin genes in ASMC was significantly higher after 24 h of co-culture with eosinophils from asthmatic subjects, while co-culture of ASMC with eosinophils from healthy subjects increased only TGF-β₁ and fibronectin gene expression. ASMC proliferation was augmented after co-culture with eosinophils from asthma patients compared with co-culture with eosinophils from healthy subjects (p < 0.05).

Conclusions

Eosinophils enhance Wnt-5a, TGF-β₁, fibronectin, and collagen gene expression in ASMC and promote proliferation of these cells in asthma.

Trial registration

ClinicalTrials.gov Identifier: NCT02648074.

Keglowich LF, Borger P. The Three A’s in Asthma - Airway Smooth Muscle, Airway Remodeling & Angiogenesis. Open Respir Med J. 2015;9:70–80.CrossRefPubMedPubMedCentral

Possa SS et al. Eosinophilic inflammation in allergic asthma. Front Pharmacol. 2013;4:46.CrossRefPubMedPubMedCentral

Doeing DC, Solway J. Airway smooth muscle in the pathophysiology and treatment of asthma. J Appl Physiol. 2013;114(7):834–43.CrossRefPubMedPubMedCentral

Fajt ML, Wenzel SE. Asthma phenotypes and the use of biologic medications in asthma and allergic disease: The next steps toward personalized care. J Allergy Clin Immunol. 2015;135(2):299–310.CrossRefPubMed

Halayko AJ et al. Airway smooth muscle phenotype and function: interactions with current asthma therapies. Curr Drug Targets. 2006;7(5):525–40.CrossRefPubMed

Holgate ST. The bronchial epithelial origins of asthma. Chem Immunol. 2000;78:62–71.CrossRefPubMed

Bousquet J et al. Asthma. From bronchoconstriction to airways inflammation and remodeling. Am J Respir Crit Care Med. 2000;161(5):1720–45.CrossRefPubMed

Kumawat K, Koopmans T, Gosens R. beta-catenin as a regulator and therapeutic target for asthmatic airway remodeling. Expert Opin Ther Targets. 2014;18(9):1023–34.CrossRefPubMed

Yeganeh B et al. Novel non-canonical TGF-β signaling networks: Emerging roles in airway smooth muscle phenotype and function. Pulm Pharmacol Ther. 2013;26(1):50–63.CrossRefPubMed

10.

James AL et al. Airway smooth muscle hypertrophy and hyperplasia in asthma. Am J Respir Crit Care Med. 2012;185(10):1058–64.CrossRefPubMed

11.

Halwani R et al. Eosinophils Induce Airway Smooth Muscle Cell Proliferation. J Allergy Clin Immunol. 2013;33(3):595–604.

12.

Kumawat K et al. Noncanonical WNT-5A signaling regulates TGF-β-induced extracellular matrix production by airway smooth muscle cells. FASEB J. 2013;27(4):1631–43.CrossRefPubMed

13.

Johnson PR et al. Extracellular matrix proteins modulate asthmatic airway smooth muscle cell proliferation via an autocrine mechanism. J Allergy Clin Immunol. 2004;113(4):690–6.CrossRefPubMed

14.

Clevers H. Wnt/beta-catenin signaling in development and disease. Cell. 2006;127(3):469–80.CrossRefPubMed

15.

Logan CY, Nusse R. The Wnt signaling pathway in development and disease. Annu Rev Cell Dev Biol. 2004;20:781–810.CrossRefPubMed

16.

Baarsma HA, Konigshoff M, Gosens R. The WNT signaling pathway from ligand secretion to gene transcription: molecular mechanisms and pharmacological targets. Pharmacol Ther. 2013;138(1):66–83.CrossRefPubMed

17.

Dekkers BG et al. Airway structural components drive airway smooth muscle remodeling in asthma. Proc Am Thorac Soc. 2009;6(8):683–92.CrossRefPubMed

18.

Kumawat K et al. TGF-β-Activated Kinase 1 (TAK1) Signaling Regulates TGF-β-Induced WNT-5A Expression in Airway Smooth Muscle Cells via Sp1 and β-Catenin. PLoS One. 2014;9(4):e94801.CrossRefPubMedPubMedCentral

19.

Gosens R et al. Role of caveolin-1 in p42/p44 MAP kinase activation and proliferation of human airway smooth muscle. Am J Physiol Lung Cell Mol Physiol. 2006;291(3):L523–34.CrossRefPubMed

20.

White SR et al. A kinetic assay for eosinophil peroxidase activity in eosinophils and eosinophil conditioned media. J Immunol Methods. 1991;144(2):257–63.CrossRefPubMed

21.

Krug N et al. Intracellular expression and serum levels of eosinophil peroxidase (EPO) and eosinophil cationic protein in asthmatic children. Clin Exp Allergy. 1999;29(11):1507–15.CrossRefPubMed

22.

Bousquet J et al. Eosinophilic inflammation in asthma. N Engl J Med. 1990;323(15):1033–9.CrossRefPubMed

23.

Bentley JK, Hershenson MB. Airway smooth muscle growth in asthma: proliferation, hypertrophy, and migration. Proc Am Thorac Soc. 2008;5(1):89–96.CrossRefPubMedPubMedCentral

24.

Kato M et al. Eosinophil infiltration and degranulation in normal human tissue. Anat Rec. 1998;252(3):418–25.CrossRefPubMed

25.

Possa SS et al. Rho-kinase inhibition attenuates airway responsiveness, inflammation, matrix remodeling, and oxidative stress activation induced by chronic inflammation. Am J Physiol Lung Cell Mol Physiol. 2012;303(11):L939–52.CrossRefPubMed

26.

Taki F et al. Effects of Rho‐kinase inactivation on eosinophilia and hyper‐reactivity in murine airways by allergen challenges. Clin Exp Allergy. 2007;37(4):599–607.CrossRefPubMed

27.

Hughes JM et al. Human eosinophil-airway smooth muscle cell interactions. Mediators Inflamm. 2000;9(2):93–9.CrossRefPubMedPubMedCentral

28.

Sur S et al. Eosinophilic inflammation is associated with elevation of interleukin-5 in the airways of patients with spontaneous symptomatic asthma. J Allergy Clin Immunol. 1995;96(5):661–8.CrossRefPubMed

29.

Mattes J et al. Intrinsic defect in T cell production of interleukin (IL)-13 in the absence of both IL-5 and eotaxin precludes the development of eosinophilia and airways hyperreactivity in experimental asthma. J Exp Med. 2002;195(11):1433–44.CrossRefPubMedPubMedCentral

30.

Lamkhioued B et al. Increased expression of eotaxin in bronchoalveolar lavage and airways of asthmatics contributes to the chemotaxis of eosinophils to the site of inflammation. J Immunol. 1997;159(9):4593–601.PubMed

31.

Lintomen L et al. Human eosinophil adhesion and degranulation stimulated with eotaxin and RANTES in vitro: lack of interaction with nitric oxide. BMC Pulmonary Med. 2008;8(1):13.CrossRef

32.

Kay AB, Phipps S, Robinson DS. A role for eosinophils in airway remodelling in asthma. Trends Immunol. 2004;25(9):477–82.CrossRefPubMed

33.

Moon RT et al. Xwnt-5A: a maternal Wnt that affects morphogenetic movements after overexpression in embryos of Xenopus laevis. Development. 1993;119(1):97–111.PubMed

34.

Sokol S. A role for Wnts in morpho-genesis and tissue polarity. Nat Cell Biol. 2000;2(7):E124–5.CrossRefPubMed

35.

Choy DF et al. Gene expression patterns of Th2 inflammation and intercellular communication in asthmatic airways. J Immunol. 2011;186(3):1861–9.CrossRefPubMed

36.

Liu L et al. Triple role of platelet-activating factor in eosinophil migration across monolayers of lung epithelial cells: eosinophil chemoattractant and priming agent and epithelial cell activator. J Immunol. 1998;161(6):3064–70.PubMed

37.

Liu L et al. Priming of eosinophil migration across lung epithelial cell monolayers and upregulation of CD11b/CD18 are elicited by extracellular Ca2+. Am J Respir Cell Mol Biol. 2003;28(6):713–21.CrossRefPubMed

Title: Eosinophils enhance WNT-5a and TGF-β1 genes expression in airway smooth muscle cells and promote their proliferation by increased extracellular matrix proteins production in asthma
Authors: Andrius Januskevicius
Simona Vaitkiene
Reinoud Gosens
Ieva Janulaityte
Deimante Hoppenot
Raimundas Sakalauskas
Kestutis Malakauskas
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: BMC Pulmonary Medicine / Issue 1/2016
Electronic ISSN: 1471-2466
DOI: https://doi.org/10.1186/s12890-016-0254-9

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Eosinophils enhance WNT-5a and TGF-β₁ genes expression in airway smooth muscle cells and promote their proliferation by increased extracellular matrix proteins production in asthma

Abstract

Background

Methods

Results

Conclusions

Trial registration

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Abstract

Background

Methods

Results

Conclusions

Trial registration

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