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Interleukin-1β Mediates β-Catenin-Driven Downregulation of Claudin-3 and Barrier Dysfunction in Caco2 Cells

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Abstract

Background

IL-1β is a cytokine involved in mediating epithelial barrier dysfunction in the gut. It is known that IL-1β mediates activation of non-muscle myosin light chain kinase in epithelial cells, but the precise mechanism by which epithelial barrier dysfunction is induced by IL-1β is not understood.

Methods and Results

Using a Caco2 cell model, we show that the expression of the tight junction protein, claudin-3, is transcriptionally downregulated by IL-1β treatment. In addition, after assessing protein and mRNA expression, and protein localization, we show that inhibition of nmMLCK rescues IL-1β-mediated decrease in claudin-3 expression as well as junction protein redistribution. Using chromatin immunoprecipitation assays, we also show that β-catenin targeting of the claudin-3 promoter occurs as a consequence of IL-1β-mediated epithelial barrier dysfunction, and inhibition of nmMLCK interferes with this interaction.

Conclusions

Taken together, these data represent the first line of evidence demonstrating nmMLCK regulation of claudin-3 expression in response to IL-1β-treated epithelial cells.

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References

  1. Ait-Belgnaoui A, Durand H, Cartier C, et al. Prevention of gut leakiness by a probiotic treatment leads to attenuated HPA response to an acute psychological stress in rats. Psychoneuroendocrinology. 2012;37:1885–1895.

    Article  CAS  PubMed  Google Scholar 

  2. Al-Sadi R, Ye D, Dokladny K, Ma TY. Mechanism of IL-1β-induced increase in intestinal epithelial tight junction permeability. J Immunol. 2008;180:5653–5661.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Arrieta MC, Madsen K, Doyle J, Meddings J. Reducing small intestinal permeability attenuates colitis in the IL10 gene-deficient mouse. Gut. 2009;58:41–48.

    Article  CAS  PubMed  Google Scholar 

  4. Beard RS, Haines RJ, Wu KY, et al. Non-muscle Mlck is required for beta-catenin- and FoxO1-dependent downregulation of Cldn5 in IL-1 beta-mediated barrier dysfunction in brain endothelial cells. J Cell Sci. 2014;127:1840–1853.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Capaldo CT, Macara IG. Depletion of E-cadherin disrupts establishment but not maintenance of cell junctions in Madin–Darby canine kidney epithelial cells. Mol Biol Cell. 2007;18:189–200.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Clayburgh DR, Rosen S, Witkowski ED, et al. A differentiation-dependent splice variant of myosin light chain kinase, MLCK1, regulates epithelial tight junction permeability. J Biol Chem. 2004;279:55506–55513.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Garcia JG, Schaphorst KL, Verin AD, Vepa S, Patterson CE, Natarajan V. Diperoxovanadate alters endothelial cell focal contacts and barrier function: role of tyrosine phosphorylation. J Appl Physiol. 2000;89:2333–2343.

    CAS  PubMed  Google Scholar 

  8. Guttman JA, Li Y, Wickham ME, Deng W, Vogl AW, Finlay BB. Attaching and effacing pathogen-induced tight junction disruption in vivo. Cell Microbiol. 2006;8:634–645.

    Article  CAS  PubMed  Google Scholar 

  9. Hashimoto K, Oshima T, Tomita T, et al. Oxidative stress induces gastric epithelial permeability through claudin-3. Biochem Biophys Res Commun. 2008;376:154–157.

    Article  CAS  PubMed  Google Scholar 

  10. Huang Q, Xu W, Ustinova E, et al. Myosin light chain kinase-dependent microvascular hyperpermeability in thermal injury. Shock. 2003;20:363–368.

    Article  CAS  PubMed  Google Scholar 

  11. Ikenouchi J, Matsuda M, Furuse M, Tsukita S. Regulation of tight junctions during the epithelium-mesenchyme transition: direct repression of the gene expression of claudins/occludin by snail. J Cell Sci. 2003;116:1959–1967.

    Article  CAS  PubMed  Google Scholar 

  12. Liebner S, Corada M, Bangsow T, et al. Wnt/β-catenin signaling controls development of the blood–brain barrier. J Cell Sci Biol. 2008;183:409–417.

    Article  CAS  Google Scholar 

  13. Ma TY, Boivin MA, Ye D, Pedram A, Said HM. Mechanism of TNF-{alpha} modulation of Caco-2 intestinal epithelial tight junction barrier: role of myosin light-chain kinase protein expression. Am J Physiol Gastrointest Liver Physiol. 2005;288:G422–430.

    Article  CAS  PubMed  Google Scholar 

  14. Madsen KL, Malfair D, Gray D, Doyle JS, Jewell LD, Fedorak RN. Interleukin-10 gene-deficient mice develop a primary intestinal permeability defect in response to enteric microflora. Inflamm Bowel Dis. 1999;5:262–270.

    Article  CAS  PubMed  Google Scholar 

  15. McLaughlin J, Padfield PJ, Burt JP, O’Neill CA. Ochratoxin A increases permeability through tight junctions by removal of specific claudin isoforms. Am J Physiol Cell Physiol. 2004;287:C1412–1417.

    Article  CAS  PubMed  Google Scholar 

  16. Mennigen R, Nolte K, Rijcken E, et al. Probiotic mixture VSL#3 protects the epithelial barrier by maintaining tight junction protein expression and preventing apoptosis in a murine model of colitis. Am J Physiol Gastrointest Liver Physiol. 2009;296:G1140–1149.

    Article  CAS  PubMed  Google Scholar 

  17. Merilainen S, Makela J, Koivukangas V, et al. Intestinal bacterial translocation and tight junction structure in acute porcine pancreatitis. Hepatogastroenterology. 2012;59:599–606.

    PubMed  Google Scholar 

  18. Mittal RD, Bid HK, Ghoshal UC. IL-1 receptor antagonist (IL-1Ra) gene polymorphism in patients with inflammatory bowel disease in India. Scand J Gastroenterol. 2005;40:827–831.

    Article  PubMed  Google Scholar 

  19. Nalle SC, Turner JR. Intestinal barrier loss as a critical pathogenic link between inflammatory bowel disease and graft-versus-host disease. Mucosal Immunol. 2015;8:720–730.

    Article  CAS  PubMed  Google Scholar 

  20. Neunlist M, Van Landeghem L, Mahe MM, Derkinderen P, des Varannes SB, Rolli-Derkinderen M. The digestive neuronal-glial-epithelial unit: a new actor in gut health and disease. Nat Rev Gastroenterol Hepatol. 2013;10:90–100.

    Article  CAS  PubMed  Google Scholar 

  21. Nighot PK, Hu CA, Ma TY. Autophagy enhances intestinal epithelial tight junction barrier function by targeting claudin-2 protein degradation. J Biol Chem. 2015;290:7234–7246.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Pinton P, Nougayrede JP, Del Rio JC, et al. The food contaminant deoxynivalenol, decreases intestinal barrier permeability and reduces claudin expression. Toxicol Appl Pharmacol. 2009;237:41–48.

    Article  CAS  PubMed  Google Scholar 

  23. Prasad S, Mingrino R, Kaukinen K, et al. Inflammatory processes have differential effects on claudins 2, 3 and 4 in colonic epithelial cells. Lab Invest. 2005;85:1139–1162.

    Article  CAS  PubMed  Google Scholar 

  24. Reinecker HC, Steffen M, Witthoeft T, et al. Enhanced secretion of tumour necrosis factor-alpha, IL-6, and IL-1 beta by isolated lamina propria mononuclear cells from patients with ulcerative colitis and Crohn’s disease. Clin Exp Immunol. 1993;94:174–181.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Reynoso R, Perrin RM, Breslin JW, et al. A role for long chain myosin light chain kinase (MLCK-210) in microvascular hyperpermeability during severe burns. Shock. 2007;28:589–595.

    CAS  PubMed  Google Scholar 

  26. Rigor RR, Shen Q, Pivetti CD, Wu MH, Yuan SY. Myosin light chain kinase signaling in endothelial barrier dysfunction. Med Res Rev. 2013;33:911–933.

    Article  CAS  PubMed  Google Scholar 

  27. Solanas G, Porta-de-la-Riva M, Agusti C, et al. E-cadherin controls beta-catenin and NF-kappaB transcriptional activity in mesenchymal gene expression. J Cell Sci. 2008;121:2224–2234.

    Article  CAS  PubMed  Google Scholar 

  28. Sun C, Wu MH, Guo M, Day ML, Lee ES, Yuan SY. ADAM15 regulates endothelial permeability and neutrophil migration via Src/ERK1/2 signalling. Cardiovasc Res. 2010;87:348–355.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Turner JR. Intestinal mucosal barrier function in health and disease. Nat Rev Immunol. 2009;9:799–809.

    Article  CAS  PubMed  Google Scholar 

  30. Verin AD, Lazar V, Torry RJ, Labarrere CA, Patterson CE, Garcia JG. Expression of a novel high molecular-weight myosin light chain kinase in endothelium. Am J Respir Cell Mol Biol. 1998;19:758–766.

    Article  CAS  PubMed  Google Scholar 

  31. Wang F, Graham WV, Wang Y, Witkowski ED, Schwarz BT, Turner JR. Interferon-gamma and tumor necrosis factor-alpha synergize to induce intestinal epithelial barrier dysfunction by up-regulating myosin light chain kinase expression. Am J Pathol. 2005;166:409–419.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Xu J, Gao XP, Ramchandran R, Zhao YY, Vogel SM, Malik AB. Nonmuscle myosin light-chain kinase mediates neutrophil transmigration in sepsis-induced lung inflammation by activating beta2 integrins. Nat Immunol. 2008;9:880–886.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Yuan SY, Wu MH, Ustinova EE, et al. Myosin light chain phosphorylation in neutrophil-stimulated coronary microvascular leakage. Circ Res. 2002;90:1214–1221.

    Article  CAS  PubMed  Google Scholar 

  34. Zeissig S, Burgel N, Gunzel D, et al. Changes in expression and distribution of claudin 2, 5 and 8 lead to discontinuous tight junctions and barrier dysfunction in active Crohn’s disease. Gut. 2007;56:61–72.

    Article  CAS  PubMed  Google Scholar 

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Disclosure of funding

Funding was provided by US Department of Veterans’ Affairs (Merit Review BX000799) and National Institutes of Health (R01 HL096640 & HL120954).

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Authors and Affiliations

  1. Department of Surgery, University of South Florida, Morsani College of Medicine, 12901 Bruce B. Downs Blvd., MDC 8, Office MDC 2012, Tampa, FL, 33612, USA

    R. J. Haines, L. Chen, R. A. Eitnier & M. H. Wu

  2. Department of Molecular Pharmacology and Physiology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA

    R. S. Beard Jr

  3. James A. Haley Veterans’ Hospital, Tampa, FL, USA

    R. J. Haines & M. H. Wu

Authors
  1. R. J. Haines
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  2. R. S. Beard Jr
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  3. L. Chen
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  4. R. A. Eitnier
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  5. M. H. Wu
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Correspondence to M. H. Wu.

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The authors have no conflict of interest to declare. The contents of this publication do not represent the views of the Department of Veterans Affairs or the US Government.

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Haines, R.J., Beard, R.S., Chen, L. et al. Interleukin-1β Mediates β-Catenin-Driven Downregulation of Claudin-3 and Barrier Dysfunction in Caco2 Cells. Dig Dis Sci 61, 2252–2261 (2016). https://doi.org/10.1007/s10620-016-4145-y

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  • DOI: https://doi.org/10.1007/s10620-016-4145-y

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