Anti‑VEGF treatment suppresses remodeling factors and restores epithelial barrier function through the E‑cadherin/β‑catenin signaling axis in experimental asthma models

Türkeli,Ahmet; Yilmaz,Özge; Karaman,Meral; Kanik,Esra Toprak; Firinci,Fatih; İnan,Sevinç; Yüksel,Hasan

doi:10.3892/etm.2021.10121

Anti‑VEGF treatment suppresses remodeling factors and restores epithelial barrier function through the E‑cadherin/β‑catenin signaling axis in experimental asthma models

Authors:
- Ahmet Türkeli
- Özge Yilmaz
- Meral Karaman
- Esra Toprak Kanik
- Fatih Firinci
- Sevinç İnan
- Hasan Yüksel
View Affiliations

Affiliations: Department of Pediatric Allergy and Immunology, Kütahya Health Science University Medical Faculty, Kütahya 43050, Turkey, Department of Pediatric Allergy and Immunology, Celal Bayar University Medical Faculty, Manisa 45030, Turkey, Multidisciplinary Laboratory, Dokuz Eylül University Medical Faculty, Izmir 35210, Turkey, Department of Pediatric Allergy and Immunology, Dokuz Eylül University Medical Faculty, Izmir 35210, Turkey, Department of Histology and Embryology, Izmir University of Economics, Medical Faculty, Izmir 35330, Turkey
Published online on: April 29, 2021 https://doi.org/10.3892/etm.2021.10121
Article Number: 689
Copyright: © Türkeli et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Besides maintaining a physical barrier with adherens junctional (AJ) and tight junctional proteins, airway epithelial cells have important roles in modulating the inflammatory processes of allergic asthma. E‑cadherin and β‑catenin are the key AJ proteins that are involved in airway remodeling. Various mediators such as transforming growth factor‑β (TGF‑β), epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet derived growth factor (PDGF), insulin‑like growth factor (IGF), tumor necrosis factor‑α (TNF‑α) and angiogenic factors, such as vascular endothelial growth factor (VEGF), are released by the airway epithelium in allergic asthma. The signaling pathways activated by these growth factors trigger epithelial‑mesenchymal transition (EMT), which contributes to fibrosis and subsequent downregulation of E‑cadherin. The present study used a mouse asthma model to investigate the effects of anti‑VEGF, anti‑TNF and corticosteroid therapies on growth factor and E‑cadherin/β‑catenin expression. The study used 38 male BALB/c mice, divided into 5 groups. A chronic mouse asthma model was created by treating 4 of the groups with inhaled and intraperitoneal ovalbumin (n= 8 per group). Saline, anti‑TNF‑α (etanercept), anti‑VEGF (bevacizumab) or a corticosteroid (dexamethasone) were applied to each group by intraperitoneal injection. No medication was administered to the control group (n=6). Immunohistochemistry for E‑cadherin, β‑catenin and growth factors was performed on lung tissues and protein expression levels assessed using H‑scores. Statistically significant differences were observed in E‑cadherin, β‑catenin, EGF, FG, and PFGF (P<0.001 for all) as well as the IGF H‑scores between the five groups (P<0.005). Only anti‑VEGF treatment caused E‑cadherin and β‑catenin levels to increase to the level of non‑asthmatic control groups (P>0.005). All treatment groups had reduced TGF‑β, PDGF and FGF H‑scores in comparison with the untreated asthma group (P=0.001). The EGF and IGF levels were not significantly different between the untreated asthmatic and non‑asthmatic controls. The results suggested that anti‑VEGF and TNF‑α inhibition treatments are effective in decreasing growth factors, in a similar manner to conventional corticosteroid treatments. Anti‑VEGF and TNF inhibition therapy may be an effective treatment for remodeling in asthma while offering an alternative therapeutic option to steroid protective agents. The data suggested that anti‑VEGF treatment offered greater restoration of the epithelial barrier than both anti‑TNF‑α and corticosteroid treatment.

View Figures

View References

1	Leynaert B, Le Moual N, Neukirch C, Siroux V and Varraso R: Environmental risk factors for asthma developement. Presse Med. 48:262–273. 2019.PubMed/NCBI View Article : Google Scholar : (In French).
2	Goleva E, Berdyshev E and Leung DY: Epithelial barrier repair and prevention of allergy. J Clin Invest. 129:1463–1474. 2019.PubMed/NCBI View Article : Google Scholar
3	Lee HY, Hur J, Kim IK, Kang JY, Yoon HK, Lee SY, Kwon SS, Kim YK and Rhee CK: Effect of nintedanib on airway inflammation and remodeling in a murine chronic asthma model. Exp Lung Res. 43:187–196. 2017.PubMed/NCBI View Article : Google Scholar
4	Kim J and Remick DG: Tumor necrosis factor inhibitors for the treatment of asthma. Curr Allergy Asthma Rep. 7:151–156. 2007.PubMed/NCBI View Article : Google Scholar
5	Guntur VP and Reinero CR: The potential use of tyrosine kinase inhibitors in severe asthma. Curr Opin Allergy Clin Immunol. 12:68–75. 2012.PubMed/NCBI View Article : Google Scholar
6	Hasan NAHM, Harith HH, Israf DA and Tham CL: The differential effects of commercial specialized media on cell growth and transforming growth factor beta 1-induced epithelial-mesenchymal transition in bronchial epithelial cells. Mol Biol Rep. 47:3511–3519. 2020.PubMed/NCBI View Article : Google Scholar
7	Yuksel H and Türkeli A: Airway epithelial barrier dysfunction in the pathogenesis and prognosis of respiratory tract diseases in childhood and adulthood. Tissue Barriers. 5(e1367458)2017.PubMed/NCBI View Article : Google Scholar
8	Chu S, Zhang X, Sun Y, Liang Y, Sun J, Lu M, Huang J, Jiang M and Ma L: Atrial natriuretic peptide inhibits epithelial-mesenchymal transition (EMT) of bronchial epithelial cells through cGMP/PKG signaling by targeting Smad3 in a murine model of allergic asthma. Exp Lung Res. 45:245–254. 2019.PubMed/NCBI View Article : Google Scholar
9	Zhang J and Dong L: Status and prospects: Personalized treatment and biomarker for airway remodeling in asthma. J Thorac Dis. 12:6090–6101. 2020.PubMed/NCBI View Article : Google Scholar
10	Hellings PW and Steelant B: Epithelial barriers in allergy and asthma. J Allergy Clin Immunol. 145:1499–1509. 2020.PubMed/NCBI View Article : Google Scholar
11	Post S, Nawijn MC, Jonker MR, Kliphuis N, van den Berge M, van Oosterhout AJ and Heijink IH: House dust mite-induced calcium signaling instigates epithelial barrier dysfunction and CCL20 production. Allergy. 68:1117–1125. 2013.PubMed/NCBI View Article : Google Scholar
12	Niessen CM: Tight junctions/adherens junctions: Basic structure and function. J Invest Dermatol. 127:2525–2532. 2007.PubMed/NCBI View Article : Google Scholar
13	Knight DA, Stick SM and Hackett TL: Defective function at the epithelial junction: A novel therapeutic frontier in asthma? J Allergy Clin Immunol. 128:557–558. 2011.PubMed/NCBI View Article : Google Scholar
14	Heijink IH, Postma DS, Noordhoek JA, Broekema M and Kapus A: House dust mite-promoted epithelial-to-mesenchymal transition in human bronchial epithelium. Am J Respir Cell Mol Biol. 42:69–79. 2010.PubMed/NCBI View Article : Google Scholar
15	Cao N, Wang J, Xu X, Xiang M and Dou J: PACAP38 improves airway epithelial barrier destruction induced by house dust mites allergen. Immunobiology. 224:758–764. 2019.PubMed/NCBI View Article : Google Scholar
16	Yao L, Chen S, Tang H, Huang P, Wei S, Liang Z, Chen X, Yang H, Tao A, Chen R, et al: Transient receptor potential ion channels mediate adherens junctions dysfunction in a toluene diisocyanate-induced murine asthma model. Toxicol Sci. 168:160–170. 2019.PubMed/NCBI View Article : Google Scholar : Erratum in Toxicol Sci 170, 247, 2019.
17	de Boer WI, Sharma HS, Baelemans SM, Hoogsteden HC, Lambrecht BN and Braunstahl GJ: Altered expression of epithelial junctional proteins in atopic asthma: Possible role in inflammation. Can J Physiol Pharmacol. 86:105–112. 2008.PubMed/NCBI View Article : Google Scholar
18	Sekiyama A, Gon Y, Terakado M, Takeshita I, Kozu Y, Maruoka S, Matsumoto K and Hashimoto S: Glucocorticoids enhance airway epithelial barrier integrity. Int Immunopharmacol. 12:350–357. 2012.PubMed/NCBI View Article : Google Scholar
19	McLellan K, Shields M, Power U and Turner S: Primary airway epithelial cell culture and asthma in children-lessons learnt and yet to come. Pediatr Pulmonol. 50:1393–1405. 2015.PubMed/NCBI View Article : Google Scholar
20	Takahashi S: Vascular endothelial growth factor (VEGF), VEGF receptors and their inhibitors for antiangiogenic tumor therapy. Biol Pharm Bull. 34(1785e1798)2011.PubMed/NCBI View Article : Google Scholar
21	Hur GY and Broide DH: Genes and pathways regulating decline in lung function and airway remodeling in asthma. Allergy Asthma Immunol Res. 11:604–621. 2019.PubMed/NCBI View Article : Google Scholar
22	Huang C, Dong H, Zou M, Luo L, Hu Y, Xie Z, Le Y, Liu L, Zou F and Cai S: Bevacizumab reduced auto-phosphorylation of VEGFR₂ to protect HDM-induced asthma mice. Biochem Biophys Res Commun. 478:181–186. 2016.PubMed/NCBI View Article : Google Scholar
23	Herbert C, Hettiaratchi A, Webb DC, Thomas PS, Foster PS and Kumar RK: Suppression of cytokine expression by roflumilast and dexamethasone in a model of chronic asthma. Clin Exp Allergy. 38:847–856. 2008.PubMed/NCBI View Article : Google Scholar
24	Ghebre MA, Pang PH, Desai D, Hargadon B, Newby C, Woods J, Rapley L, Cohen SE, Herath A, Gaillard EA, et al: Severe exacerbations in moderate-to-severe asthmatics are associated with increased pro-inflammatory and type 1 mediators in sputum and serum. BMC Pulm Med. 19(144)2019.PubMed/NCBI View Article : Google Scholar
25	Malaviya R, Laskin JD and Laskin DL: Anti-TNFα therapy in inflammatory lung diseases. Pharmacol Ther. 180:90–98. 2017.PubMed/NCBI View Article : Google Scholar
26	Mukhopadhyay S, Hoidal JR and Mukherjee TK: Role of TNFalpha in pulmonary pathophysiology. Respir Res. 7(125)2006.PubMed/NCBI View Article : Google Scholar
27	National Research Council (US) Committee for the Update of the Guide for the Care and Use of Laboratory Animal: Guide for the Care and Use of Laboratory Animals, 8th edition. National Academies Press (US), Washington, DC, 2011.
28	Temelkovski J, Hogan SP, Shepherd DP, Foster PS and Kumar RK: An improved murine model of asthma: Selective airway inflammation, epithelial lesions and increased methacholine responsiveness following chronic exposure to aerosolised allergen. Thorax. 53:849–856. 1998.PubMed/NCBI View Article : Google Scholar
29	Cardiff RD, Miller CH and Munn RJ: Manual hematoxylin and eosin staining of mouse tissue sections. Cold Spring Harb Protoc. 2014:655–658. 2014.PubMed/NCBI View Article : Google Scholar
30	Yuksel H, Yilmaz O, Baytur YB and Ozbilgin K: Prenatal administration of granulocyte-macrophage colony-stimulating factor increases mesenchymal vascular endothelial growth factor expression and maturation in fetal rat lung. Exp Lung Res. 34:550–558. 2008.PubMed/NCBI View Article : Google Scholar
31	Wang T, Zhou Q and Shang Y: MiRNA-451a inhibits airway remodeling by targeting Cadherin 11 in an allergic asthma model of neonatal mice. Int Immunopharmacol. 83(106440)2020.PubMed/NCBI View Article : Google Scholar
32	Gong JH, Cho IH, Shin D, Han SY, Park SH and Kang YH: Inhibition of airway epithelial-to-mesenchymal transition and fibrosis by kaempferol in endotoxin-induced epithelial cells and ovalbumin-sensitized mice. Lab Invest. 94:297–308. 2014.PubMed/NCBI View Article : Google Scholar
33	Makinde T, Murphy RF and Agrawal DK: Immunomodulatory role of vascular endothelial growth factor and angiopoietin-1 in airway remodeling. Curr Mol Med. 6:831–841. 2006.PubMed/NCBI View Article : Google Scholar
34	Hough KP, Curtiss ML, Blain TJ, Liu RM, Trevor J, Deshane JS and Thannickal VJ: Airway remodeling in asthma. Front Med (Lausanne). 7(191)2020.PubMed/NCBI View Article : Google Scholar
35	Hasan NAHM, Harith HH, Israf DA and Tham CL: The differential effects of commercial specialized media on cell growth and transforming growth factor beta 1-induced epithelial-mesenchymal transition in bronchial epithelial cells. Mol Biol Rep. 47:3511–3519. 2020.PubMed/NCBI View Article : Google Scholar
36	Frey A, Lunding LP, Ehlers JC, Weckmann M, Zissler UM and Wegmann M: More than just a barrier: The immune functions of the airway epithelium in asthma pathogenesis. Front Immunol. 11(761)2020.PubMed/NCBI View Article : Google Scholar
37	Post S, Heijink IH, Hesse L, Koo HK, Shaheen F, Fouadi M, Kuchibhotla VNS, Lambrecht BN, Van Oosterhout AJM, Hackett TL, et al: Characterization of a lung epithelium specific E-cadherin knock-out model: Implications for obstructive lung pathology. Sci Rep. 8(13275)2018.PubMed/NCBI View Article : Google Scholar
38	Cai J, Culley MK, Zhao Y and Zhao J: The role of ubiquitination and deubiquitination in the regulation of cell junctions. Protein Cell. 9:754–769. 2018.PubMed/NCBI View Article : Google Scholar
39	Goto Y, Uchida Y, Nomura A, Sakamoto T, Ishii Y, Morishima Y, Masuyama K and Sekizawa K: Dislocation of E-cadherin in the airway epithelium during an antigen-induced asthmatic response. Am J Respir Cell Mol Biol. 23:712–718. 2000.PubMed/NCBI View Article : Google Scholar
40	Baarsma HA, Menzen MH, Halayko AJ, Meurs H, Kerstjens HA and Gosens R: β-catenin signaling is required for TGF-β1-induced extracellular matrix production by airway smooth muscle cells. Am J Physiol Lung Cell Mol Physiol. 301:L956–L965. 2011.PubMed/NCBI View Article : Google Scholar
41	Kim HT, Yin W, Nakamichi Y, Panza P, Grohmann B, Buettner C, Guenther S, Ruppert C, Kobayashi Y, Guenther A, et al: WNT/RYK signaling restricts goblet cell differentiation during lung development and repair. Proc Natl Acad Sci USA. 116:25697–25706. 2019.PubMed/NCBI View Article : Google Scholar
42	Jia XX, Zhu TT, Huang Y, Zeng XX, Zhang H and Zhang WX: Wnt/β-catenin signaling pathway regulates asthma airway remodeling by influencing the expression of c-Myc and cyclin D1 via the p38 MAPK-dependent pathway. Exp Ther Med. 18:3431–3438. 2019.PubMed/NCBI View Article : Google Scholar
43	Evans SM, Blyth DI, Wong T, Sanjar S and West MR: Decreased distribution of lung epithelial junction proteins after intratracheal antigen or lipopolysaccharide challenge: Correlation with neutrophil influx and levels of BALF sE-cadherin. Am J Respir Cell Mol Biol. 27:446–454. 2002.PubMed/NCBI View Article : Google Scholar
44	Xiao C, Puddicombe SM, Field S, Haywood J, Broughton-Head V, Puxeddu I, Haitchi HM, Vernon-Wilson E, Sammut D, Bedke N, et al: Defective epithelial barrier function in asthma. J Allergy Clin Immunol. 128:549–56.e1-12. 2011.PubMed/NCBI View Article : Google Scholar
45	Trautmann A, Kruger K, Akdis M, Muller-Wening D, Akkaya A, Brocker EB, Blaser K and Akdis CA: Apoptosis and loss of adhesion of bronchial epithelial cells in asthma. Int Arch Allergy Immunol. 138:142–150. 2005.PubMed/NCBI View Article : Google Scholar
46	Yuksel H, Türkeli A, Taneli F, Horasan GD, Kanik ET, Kizilkaya M, Gözükara C and Yilmaz O: E-cadherin as an epithelial barrier protein in exhaled breath condensate. J Breath Res. 8(046006)2014.PubMed/NCBI View Article : Google Scholar
47	Willems-Widyastuti A, Vanaudenaerde BM, Vos R, Dilisen E, Verleden SE, De Vleeschauwer SI, Vaneylen A, Mooi WJ, de Boer WI, Sharma HS, et al: Azithromycin attenuates fibroblast growth factors induced vascular endothelial growth factor via p38(MAPK) signaling in human airway smooth muscle cells. Cell Biochem Biophys. 67:331–339. 2013.PubMed/NCBI View Article : Google Scholar
48	Carayol N, Campbell A, Vachier I, Mainprice B, Bousquet J, Godard P and Chanez P: Modulation of cadherin and catenins expression by tumor necrosis factor-alpha and dexamethasone in human bronchial epithelial cells. Am J Respir Cell Mol Biol. 26:341–347. 2002.PubMed/NCBI View Article : Google Scholar
49	Moheimani F, Roth HM, Cross J, Reid AT, Shaheen F, Warner SM, Hirota JA, Kicic A, Hallstrand TS, Kahn M, et al: Disruption of β-catenin/CBP signaling inhibits human airway epithelial-mesenchymal transition and repair. Int J Biochem Cell Biol. 68:59–69. 2015.PubMed/NCBI View Article : Google Scholar
50	Winton HL, Wan H, Cannell MB, Thompson PJ, Garrod DR, Stewart GA and Robinson C: Class specific inhibition of house dust mite proteinases which cleave cell adhesion, induce cell death and which increase the permeability of lung epithelium. Br J Pharmacol. 124:1048–1059. 1998.PubMed/NCBI View Article : Google Scholar
51	Park HY, Kim JH and Park CK: VEGF induces TGF-β1 expression and myofibroblast transformation after glaucoma surgery. Am J Pathol. 182:2147–2154. 2013.PubMed/NCBI View Article : Google Scholar
52	Chatterjee S, Wang Y, Duncan MK and Naik UP: Junctional adhesion molecule-A regulates vascular endothelial growth factor receptor-2 signaling-dependent mouse corneal wound healing. PLoS One. 8(e63674)2013.PubMed/NCBI View Article : Google Scholar
53	Wallez Y and Huber P: Endothelial adherens and tight junctions in vascular homeostasis, inflammation and angiogenesis. Biochim Biophys Acta. 1778:794–809. 2008.PubMed/NCBI View Article : Google Scholar
54	Díaz-Coránguez M, Lin CM, Liebner S and Antonetti DA: Norrin restores blood-retinal barrier properties after vascular endothelial growth factor-induced permeability. J Biol Chem. 295:4647–4660. 2020.PubMed/NCBI View Article : Google Scholar
55	He J, Wang H, Liu Y, Li W, Kim D and Huang H: Blockade of vascular endothelial growth factor receptor 1 prevents inflammation and vascular leakage in diabetic retinopathy. J Ophthalmol. 2015(605946)2015.PubMed/NCBI View Article : Google Scholar
56	Wisniewska-Kruk J, Hoeben KA, Vogels IM, Gaillard PJ, Van Noorden CJ, Schlingemann RO and Klaassen I: A novel co-culture model of the blood-retinal barrier based on primary retinal endothelial cells, pericytes and astrocytes. Exp Eye Res. 96:181–190. 2012.PubMed/NCBI View Article : Google Scholar : Erratum in Exp Eye Res 138, 167, 2015.
57	Harhaj NS, Felinski EA, Wolpert EB, Sundstrom JM, Gardner TW and Antonetti DA: VEGF activation of protein kinase C stimulates occludin phosphorylation and contributes to endothelial permeability. Invest Ophthalmol Vis Sci. 47:5106–5115. 2006.PubMed/NCBI View Article : Google Scholar
58	Sullivan DE, Ferris M, Nguyen H, Abboud E and Brody AR: TNF-alpha induces TGF-beta1 expression in lung fibroblasts at the transcriptional level via AP-1 activation. J Cell Mol Med. 13:1866–1876. 2009.PubMed/NCBI View Article : Google Scholar
59	Brightling C, Berry M and Amrani Y: Targeting TNF-alpha: A novel therapeutic approach for asthma. J Allergy Clin Immunol. 121:5–10; quiz 11-12. 2008.PubMed/NCBI View Article : Google Scholar
60	Câmara J and Jarai G: . Epithelial-mesenchymal transition in primary human bronchial epithelial cells is Smad-dependent and enhanced by fibronectin and TNF-alpha. Fibrogenesis Tissue Repair. 3(2)2010.PubMed/NCBI View Article : Google Scholar
61	Hardyman MA, Wilkinson E, Martin E, Jayasekera NP, Blume C, Swindle EJ, Gozzard N, Holgate ST, Howarth PH, Davies DE, et al: TNF-α-mediated bronchial barrier disruption and regulation by src-family kinase activation. J Allergy Clin Immunol. 132:665–675.e8. 2013.PubMed/NCBI View Article : Google Scholar
62	Coyne CB, Vanhook MK, Gambling TM, Carson JL, Boucher RC and Johnson LG: Regulation of airway tight junctions by proinflammatory cytokines. Mol Biol Cell. 13:3218–3234. 2002.PubMed/NCBI View Article : Google Scholar
63	Pohl C, Hermanns MI, Uboldi C, Bock M, Fuchs S, Dei-Anang J, Mayer E, Kehe K, Kummer W and Kirkpatrick CJ: Barrier functions and paracellular integrity in human cell culture models of the proximal respiratory unit. Eur J Pharm Biopharm. 72:339–349. 2009.PubMed/NCBI View Article : Google Scholar
64	Doherty T and Broide D: Cytokines and growth factors in airway remodeling in asthma. Curr Opin Immunol. 19:676–680. 2007.PubMed/NCBI View Article : Google Scholar
65	Wang Z, Li W, Guo Q, Wang Y, Ma L and Zhang X: Insulin-like growth factor-1 signaling in lung development and inflammatory lung diseases. BioMed Res Int. 2018(6057589)2018.PubMed/NCBI View Article : Google Scholar
66	Dekkers BG, Pehlic A, Mariani R, Bos IS, Meurs H and Zaagsma J: Glucocorticosteroids and β₂-adrenoceptor agonists synergize to inhibit airway smooth muscle remodeling. J Pharmacol Exp Ther. 342:780–787. 2012.PubMed/NCBI View Article : Google Scholar
67	Song J, Zhao H, Dong H, Zhang D, Zou M, Tang H, Liu L, Liang Z, Lv Y, Zou F, et al: Mechanism of E-cadherin redistribution in bronchial airway epithelial cells in a TDI-induced asthma model. Toxicol Lett. 220:8–14. 2013.PubMed/NCBI View Article : Google Scholar
68	Doerner AM and Zuraw BL: TGF-β1 induced epithelial to mesenchymal transition (EMT) in human bronchial epithelial cells is enhanced by IL-1β but not abrogated by corticosteroids. Respir Res. 10(100)2009.PubMed/NCBI View Article : Google Scholar
69	Kimura K, Teranishi S, Kawamoto K and Nishida T: Protective effect of dexamethasone against hypoxia-induced disruption of barrier function in human corneal epithelial cells. Exp Eye Res. 92:388–393. 2011.PubMed/NCBI View Article : Google Scholar