Advertisement

Interference of miR-943-3p with secreted frizzled-related proteins4 (SFRP4) in an asthma mouse model

  • Jian ShenEmail author
  • Jun Zhao
  • Qing-yan Ye
  • Xi-dong Gu
Regular Article
  • 14 Downloads

Abstract

The aim of this study is to investigate the potential roles of miR-943-3p and its target gene secreted frizzled-related proteins4 (SFRP4) in allergic asthma and elucidate its underlying mechanism, which may prompt a new clue about developing novel treatments of this disease. An allergic asthma mouse model was generated by challenging with ovalbumin (OVA); lung pathological features of mice were viewed using H&E staining; thickness of subepithelial fibrosis and smooth muscle was measured using Masson’s trichrome staining. Inflammatory cells from bronchoalveolar lavage fluid (BALF) were counted based on Diff-Quik staining and morphometric analysis. Expressions of miR-943-3p, SFRP4 and Wnt signal pathway-associated proteins were detected using RT-PCR or immunoblotting, respectively. SFRP4 was downregulated in the bronchial biopsies of allergic asthma patients and represented a unique intersection between differentially expressed genes (DEGs) and genes in the Wnt signal pathway. Both miR-943-3p upregulation and SFRP4 downregulation were detected in allergic asthma patients and OVA-induced mice. Besides, OVA-induced mice possessed more inflammatory cells in BALF including macrophage (mac), eosinophil (eos), lymphocyte (lym) and neutrophil (neu), higher expression of collagen, β-catenin and c-Myc as well as thicker subepithelial fibrosis and smooth muscle in lung than control mice. In vivo delivery of miR-943-3p agomir worsened these symptoms, while both miR-943-3p antagomir and Ad-SFRP4 administration effectively alleviated this disease. Taken together, miR-943-3p accelerated the progression of airway inflammation and remodeling in allergic asthma via suppressing the activity of SFRP4 through Wnt signaling pathway in asthma patients and OVA-induced mice.

Keywords

Allergic asthma SFRP4 miR-943-3p Wnt signaling pathway Secreted protein 

Notes

Funding

This study was supported by the Science and Technology Commission of Shanghai Municipality Traditional Chinese Medicine (TCM specialist) Specialized Personnel Plan (ZY3-RCPY-3-1027), Shanghai Special Program for Children’s Health Service Capacity Building High Pediatric Overseas Training Team Cultivation-Pediatrics of Integrated Traditional Chinese and Western Medicine (GDEK201704) and Important Subject Construction of Shanghai Health and Family Planning System-Pediatrics of Integrative Chinese and Western Medicine (2016ZB0104-02).

Compliance with ethical standards

Ethics approval and consent to participate

This study was authorized by the Shuguang Hospital Affiliated to Shanghai Traditional Chinese Medical University, who obtained written informed consents from all the participants.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Conflict of interest

The authors declare that they have no conflicts of interest.

References

  1. Andersson CK, Adams A, Nagakumar P, Bossley C, Gupta A, De Vries D, Adnan A, Bush A, Saglani S, Lloyd CM (2017) Intraepithelial neutrophils in pediatric severe asthma are associated with better lung function. J Allergy Clin Immunol 139:1819–1829 e1811CrossRefGoogle Scholar
  2. Bafico A, Gazit A, Pramila T, Finch PW, Yaniv A, Aaronson SA (1999) Interaction of frizzled related protein (FRP) with Wnt ligands and the frizzled receptor suggests alternative mechanisms for FRP inhibition of Wnt signaling. J Biol Chem 274:16180–16187CrossRefGoogle Scholar
  3. Bartel DP (2009) MicroRNAs: target recognition and regulatory functions. Cell 136:215–233CrossRefGoogle Scholar
  4. Beckert H, Meyer-Martin H, Buhl R, Taube C, Reuter S (2018) The canonical but not the noncanonical Wnt pathway inhibits the development of allergic airway disease. J Immunol 201:1855–1864Google Scholar
  5. Bijkerk R, de Bruin RG, van Solingen C, van Gils JM, Duijs JM, van der Veer EP, Rabelink TJ, Humphreys BD, van Zonneveld AJ (2016) Silencing of microRNA-132 reduces renal fibrosis by selectively inhibiting myofibroblast proliferation. Kidney Int 89:1268–1280CrossRefGoogle Scholar
  6. Carmon KS, Loose DS (2008) Secreted frizzled-related protein 4 regulates two Wnt7a signaling pathways and inhibits proliferation in endometrial cancer cells. Mol Cancer Res 6:1017–1028CrossRefGoogle Scholar
  7. Carraro G, Shrestha A, Rostkovius J, Contreras A, Chao CM, El Agha E, Mackenzie B, Dilai S, Guidolin D, Taketo MM, Gunther A, Kumar ME, Seeger W, De Langhe S, Barreto G, Bellusci S (2014) miR-142-3p balances proliferation and differentiation of mesenchymal cells during lung development. Development 141:1272–1281CrossRefGoogle Scholar
  8. Chamberland A, Madore AM, Tremblay K, Laviolette M, Laprise C (2009) A comparison of two sets of microarray experiments to define allergic asthma expression pattern. Exp Lung Res 35:399–410CrossRefGoogle Scholar
  9. Chauhan BF, Ducharme FM (2012) Anti-leukotriene agents compared to inhaled corticosteroids in the management of recurrent and/or chronic asthma in adults and children. Cochrane Database Syst Rev:CD002314Google Scholar
  10. Cohen ED, Ihida-Stansbury K, Lu MM, Panettieri RA, Jones PL, Morrisey EE (2009) Wnt signaling regulates smooth muscle precursor development in the mouse lung via a tenascin C/PDGFR pathway. J Clin Invest 119:2538–2549CrossRefGoogle Scholar
  11. Collison A, Herbert C, Siegle JS, Mattes J, Foster PS, Kumar RK (2011) Altered expression of microRNA in the airway wall in chronic asthma: miR-126 as a potential therapeutic target. BMC Pulm Med 11:29CrossRefGoogle Scholar
  12. Fehrenbach H, Wagner C, Wegmann M (2017) Airway remodeling in asthma: what really matters. Cell Tissue Res 367:551–569CrossRefGoogle Scholar
  13. Haraguchi R, Kitazawa R, Mori K, Tachibana R, Kiyonari H, Imai Y, Abe T, Kitazawa S (2016) sFRP4-dependent Wnt signal modulation is critical for bone remodeling during postnatal development and age-related bone loss. Sci Rep 6:25198CrossRefGoogle Scholar
  14. Hirota N, Martin JG (2013) Mechanisms of airway remodeling. Chest 144:1026–1032CrossRefGoogle Scholar
  15. Horvath LG, Henshall SM, Kench JG, Saunders DN, Lee CS, Golovsky D, Brenner PC, O’Neill GF, Kooner R, Stricker PD, Grygiel JJ, Sutherland RL (2004) Membranous expression of secreted frizzled-related protein 4 predicts for good prognosis in localized prostate cancer and inhibits PC3 cellular proliferation in vitro. Clin Cancer Res 10:615–625CrossRefGoogle Scholar
  16. Horvath LG, Lelliott JE, Kench JG, Lee CS, Williams ED, Saunders DN, Grygiel JJ, Sutherland RL, Henshall SM (2007) Secreted frizzled-related protein 4 inhibits proliferation and metastatic potential in prostate cancer. Prostate 67:1081–1090CrossRefGoogle Scholar
  17. Hromadnikova I, Kotlabova K, Hympanova L, Doucha J, Krofta L (2014) First trimester screening of circulating C19MC microRNAs can predict subsequent onset of gestational hypertension. PLoS One 9:e113735CrossRefGoogle Scholar
  18. Ji Q, Zhang J, Du Y, Zhu E, Wang Z, Que B, Miao H, Shi S, Qin X, Zhao Y, Zhou Y, Huang F, Nie S (2017) Human epicardial adipose tissue-derived and circulating secreted frizzled-related protein 4 (SFRP4) levels are increased in patients with coronary artery disease. Cardiovasc Diabetol 16:133CrossRefGoogle Scholar
  19. Koh YI, Shim JU, Lee JH, Chung IJ, Min JJ, Rhee JH, Lee HC, Chung DH, Wi JO (2010) Natural killer T cells are dispensable in the development of allergen-induced airway hyperresponsiveness, inflammation and remodelling in a mouse model of chronic asthma. Clin Exp Immunol 161:159–170Google Scholar
  20. Kwak HJ, Park DW, Seo JY, Moon JY, Kim TH, Sohn JW, Shin DH, Yoon HJ, Park SS, Kim SH (2015) The Wnt/beta-catenin signaling pathway regulates the development of airway remodeling in patients with asthma. Exp Mol Med 47:e198CrossRefGoogle Scholar
  21. Laffont B, Rayner KJ (2017) MicroRNAs in the pathobiology and therapy of atherosclerosis. Can J Cardiol 33:313–324CrossRefGoogle Scholar
  22. Long JW, Yang XD, Cao L, Lu SM, Cao YX (2009) Alteration of airway responsiveness mediated by receptors in ovalbumin-induced asthmatic E3 rats. Acta Pharmacol Sin 30:965–972CrossRefGoogle Scholar
  23. Ma B, Hottiger MO (2016) Crosstalk between Wnt/beta-catenin and NF-kappaB signaling pathway during inflammation. Front Immunol 7:378Google Scholar
  24. Matsushima K, Suyama T, Takenaka C, Nishishita N, Ikeda K, Ikada Y, Sawa Y, Jakt LM, Mori H, Kawamata S (2010) Secreted frizzled related protein 4 reduces fibrosis scar size and ameliorates cardiac function after ischemic injury. Tissue Eng Part A 16:3329–3341CrossRefGoogle Scholar
  25. Mortensen MM, Hoyer S, Lynnerup AS, Orntoft TF, Sorensen KD, Borre M, Dyrskjot L (2015) Expression profiling of prostate cancer tissue delineates genes associated with recurrence after prostatectomy. Sci Rep 5:16018CrossRefGoogle Scholar
  26. Munakata M (2006) Airway remodeling and airway smooth muscle in asthma. Allergol Int 55:235–243CrossRefGoogle Scholar
  27. Ober C, Yao TC (2011) The genetics of asthma and allergic disease: a 21st century perspective. Immunol Rev 242:10–30CrossRefGoogle Scholar
  28. Ogawa H, Azuma M, Muto S, Nishioka Y, Honjo A, Tezuka T, Uehara H, Izumi K, Itai A, Sone S (2011) IkappaB kinase beta inhibitor IMD-0354 suppresses airway remodelling in a Dermatophagoides pteronyssinus-sensitized mouse model of chronic asthma. Clin Exp Allergy 41:104–115CrossRefGoogle Scholar
  29. Pohl S, Scott R, Arfuso F, Perumal V, Dharmarajan A (2015) Secreted frizzled-related protein 4 and its implications in cancer and apoptosis. Tumour Biol 36:143–152CrossRefGoogle Scholar
  30. Reddel HK, Bateman ED, Becker A, Boulet LP, Cruz AA, Drazen JM, Haahtela T, Hurd SS, Inoue H, de Jongste JC, Lemanske RF Jr, Levy ML, O’Byrne PM, Paggiaro P, Pedersen SE, Pizzichini E, Soto-Quiroz M, Szefler SJ, Wong GW, FitzGerald JM (2015) A summary of the new GINA strategy: a roadmap to asthma control. Eur Respir J 46:622–639CrossRefGoogle Scholar
  31. Reuter S, Martin H, Beckert H, Bros M, Montermann E, Belz C, Heinz A, Ohngemach S, Sahin U, Stassen M, Buhl R, Eshkind L, Taube C (2014) The Wnt/beta-catenin pathway attenuates experimental allergic airway disease. J Immunol 193:485–495CrossRefGoogle Scholar
  32. Sandsmark E, Andersen MK, Bofin AM, Bertilsson H, Drablos F, Bathen TF, Rye MB, Tessem MB (2017) SFRP4 gene expression is increased in aggressive prostate cancer. Sci Rep 7:14276CrossRefGoogle Scholar
  33. Schatz M, Rosenwasser L (2014) The allergic asthma phenotype. J Allergy Clin Immunol Pract 2:645–648 quiz 649CrossRefGoogle Scholar
  34. Sol IS, Kim YH, Park YA, Lee KE, Hong JY, Kim MN, Kim YS, Oh MS, Yoon SH, Kim MJ, Kim KW, Sohn MH, Kim KE (2016) Relationship between sputum clusterin levels and childhood asthma. Clin Exp Allergy 46:688–695CrossRefGoogle Scholar
  35. Vallee A, Lecarpentier Y (2018) Crosstalk between peroxisome proliferator-activated receptor gamma and the canonical WNT/beta-catenin pathway in chronic inflammation and oxidative stress during carcinogenesis. Front Immunol 9:745CrossRefGoogle Scholar
  36. Van Scoyk M, Randall J, Sergew A, Williams LM, Tennis M, Winn RA (2008) Wnt signaling pathway and lung disease. Transl Res 151:175–180CrossRefGoogle Scholar
  37. Yang X, Lv JN, Li H, Jiao B, Zhang QH, Zhang Y, Zhang J, Liu YQ, Zhang M, Shan H, Zhang JZ, Wu RM, Li YL (2017) Curcumin reduces lung inflammation via Wnt/beta-catenin signaling in mouse model of asthma. J Asthma 54:335–340CrossRefGoogle Scholar
  38. Yao L, Zhao H, Tang H, Xiong J, Zhao W, Liu L, Dong H, Zou F, Cai S (2017) Blockade of beta-catenin signaling attenuates toluene diisocyanate-induced experimental asthma. Allergy 72:579–589CrossRefGoogle Scholar
  39. Ye L, Mou Y, Wang J, Jin ML (2017) Effects of microRNA-19b on airway remodeling, airway inflammation and degree of oxidative stress by targeting TSLP through the Stat3 signaling pathway in a mouse model of asthma. Oncotarget 8:47533–47546Google Scholar
  40. Yin H, Zhang S, Sun Y, Li S, Ning Y, Dong Y, Shang Y, Bai C (2017) MicroRNA-34/449 targets IGFBP-3 and attenuates airway remodeling by suppressing Nur77-mediated autophagy. Cell Death Dis 8:e2998CrossRefGoogle Scholar
  41. Yoshikawa T, Shoji S, Fujii T, Kanazawa H, Kudoh S, Hirata K, Yoshikawa J (1998) Severity of exercise-induced bronchoconstriction is related to airway eosinophilic inflammation in patients with asthma. Eur Respir J 12:879–884CrossRefGoogle Scholar
  42. Zein JG, Dweik RA, Comhair SA, Bleecker ER, Moore WC, Peters SP, Busse WW, Jarjour NN, Calhoun WJ, Castro M, Chung KF, Fitzpatrick A, Israel E, Teague WG, Wenzel SE, Love TE, Gaston BM, Erzurum SC, Severe Asthma Research P (2015) Asthma is more severe in older adults. PLoS One 10:e0133490CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of PediatricsShuguang Hospital Affiliated to Shanghai Traditional Chinese Medical UniversityShanghaiChina
  2. 2.Department of Clinical LaboratoryShuguang Hospital Affiliated to Shanghai Traditional Chinese Medical UniversityShanghaiChina

Personalised recommendations