Muc5ac null mice are predisposed to spontaneous gastric antro-pyloric hyperplasia and adenomas coupled with attenuated H. pylori-induced corpus mucous metaplasia

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Gastric cancer (GC) is the third leading cause of cancer-related deaths worldwide and is strongly associated with chronic Helicobacter pylori (Hp) infection. The ability of Hp to closely adhere to the gastric surface protective mucous layer containing mucins (MUC in humans and Muc in animals), primarily Muc5ac, is integral in the stepwise pathogenesis from gastritis to cancer. To probe the role of Muc5ac in Hp-induced gastric pathology, Muc5ac−/− and Muc5ac+/+ (WT) mice were experimentally infected with Hp Sydney strain (SS1). At 16 weeks and 32 weeks post infection (wpi), groups of mice were euthanized and evaluated for the following: gastric histopathological parameters, immunohistochemical expression of mucins (Muc5ac, Muc1, Muc2), Trefoil factor family proteins (Tff1 and Tff2), Griffonia (Bandeiraea) simplicifolia lectin II (GSL II) (mucous metaplasia marker) and Clusterin (Spasmolytic Polypeptide Expressing Metaplasia (SPEM) marker), Hp colonization density by qPCR and gastric cytokine mRNA levels. Our results demonstrate that Muc5ac−/− mice developed spontaneous antro-pyloric proliferation, adenomas and in one case with neuroendocrine differentiation; these findings were independent of Hp infection along with strong expression levels of Tff1, Tff2 and Muc1. Hp-infected Muc5ac−/− mice had significantly lowered gastric corpus mucous metaplasia at 16 wpi and 32 wpi (P = 0.0057 and P = 0.0016, respectively), with a slight reduction in overall gastric corpus pathology. GSII-positive mucous neck cells were decreased in Hp-infected Muc5ac−/− mice compared to WT mice and clusterin positivity was noted within metaplastic glands in both genotypes following Hp infection. Additionally, Hp colonization densities were significantly higher in Muc5ac−/− mice compared to WT at 16 wpi in both sexes (P = 0.05) along with a significant reduction in gastric Tnfα (16 wpi—males and females, P = 0.017 and P = 0.036, respectively and 32 wpi—males only, P = 0.025) and Il-17a (16 wpi—males) (P = 0.025). Taken together, our findings suggest a protective role for MUC5AC/Muc5ac in maintaining gastric antral equilibrium and inhibiting Hp colonization and associated inflammatory pathology.

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  1. 1.

    Moore ME, Boren T, Solnick JV. Life at the margins: modulation of attachment proteins in Helicobacter pylori. Gut Microbes. 2011;2:42–6.

  2. 2.

    Oleastro M, Menard A. The role of Helicobacter pylori outer membrane proteins in adherence and pathogenesis. Biology (Basel). 2013;2:1110–34.

  3. 3.

    Juge N. Microbial adhesins to gastrointestinal mucus. Trends Microbiol. 2012;20:30–9.

  4. 4.

    Yandrapu H, Sarosiek J. Protective factors of the gastric and duodenal mucosa: an overview. Curr Gastroenterol Rep. 2015;17:24.

  5. 5.

    Kemmerly T, Kaunitz JD. Gastroduodenal mucosal defense. Curr Opin Gastroenterol. 2013;29:642–9.

  6. 6.

    Hasnain SZ, Gallagher AL, Grencis RK, Thornton DJ. A new role for mucins in immunity: insights from gastrointestinal nematode infection. Int J Biochem Cell Biol. 2013;45:364–74.

  7. 7.

    Schmitz JM, Durham CG, Ho SB, Lorenz RG. Gastric mucus alterations associated with murine Helicobacter infection. J Histochem Cytochem. 2009;57:457–67.

  8. 8.

    Babu SD, Jayanthi V, Devaraj N, Reis CA, Devaraj H. Expression profile of mucins (MUC2, MUC5AC and MUC6) in Helicobacter pylori infected pre-neoplastic and neoplastic human gastric epithelium. Mol Cancer. 2006;5:10.

  9. 9.

    Machado JC, Nogueira AM, Carneiro F, Reis CA, Sobrinho-Simões M. Gastric carcinoma exhibits distinct types of cell differentiation: an immunohistochemical study of trefoil peptides (TFF1 and TFF2) and mucins (MUC1, MUC2, MUC5AC, and MUC6). J Pathol. 2000;190:437–43.

  10. 10.

    Park JS, Yeom JS, Seo JH, Lim JY, Park CH, Woo HO, et al. Immunohistochemical expressions of MUC2, MUC5AC, and MUC6 in normal, Helicobacter pylori infected and metaplastic gastric mucosa of children and adolescents. Helicobacter. 2015;20:260–8.

  11. 11.

    Liu C, Smet A, Blaecher C, Flahou B, Ducatelle R, Linden S, et al. Gastric de novo Muc13 expression and spasmolytic polypeptide-expressing metaplasia during Helicobacter heilmannii infection. Infect Immun. 2014;82:3227–39.

  12. 12.

    Teixeira A, David L, Reis CA, Costa J, Sobrinho-Simões M. Expression of mucins (MUC1, MUC2, MUC5AC, and MUC6) and type 1 Lewis antigens in cases with and without Helicobacter pylori colonization in metaplastic glands of the human stomach. J Pathol. 2002;197:37–43.

  13. 13.

    Boltin D, Niv Y. Mucins in gastric cancer—an update. J Gastrointest Dig Syst. 2013;3:15519.

  14. 14.

    Schreiber S, Konradt M, Groll C, Scheid P, Hanauer G, Werling HO, et al. The spatial orientation of Helicobacter pylori in the gastric mucus. Proc Natl Acad Sci USA. 2004;101:5024–9.

  15. 15.

    Lindén SK, Wickström C, Lindell G, Gilshenan K, Carlstedt I. Four modes of adhesion are used during Helicobacter pylori binding to human mucins in the oral and gastric niches. Helicobacter. 2008;13:81–93.

  16. 16.

    Borén T, Falk P, Roth KA, Larson G, Normark S. Attachment of Helicobacter pylori to human gastric epithelium mediated by blood group antigens. Science. 1993;262:1892–5.

  17. 17.

    Dunne C, Dolan B, Clyne M. Factors that mediate colonization of the human stomach by Helicobacter pylori. World J Gastroenterol. 2014;20:5610–24.

  18. 18.

    Van de Bovenkamp JH, Mahdavi J, Korteland-Van Male AM, Büller HA, Einerhand AW, Borén T, et al. The MUC5AC glycoprotein is the primary receptor for Helicobacter pylori in the human stomach. Helicobacter. 2003;8:521–32.

  19. 19.

    Lemke LB, Ge Z, Whary MT, Feng Y, Rogers AB, Muthupalani S, et al. Concurrent Helicobacter bilis infection attenuates proinflammatory H. pylori induced gastric pathology. Infect Immun. 2009;77:2147–58.

  20. 20.

    Ge Z, Feng Y, Muthupalani S, Eurell LL, Taylor NS, Whary MT, et al. Coinfection with Enterohepatic Helicobacter species can ameliorate or promote Helicobacter pylori-induced gastric pathology in C57BL/6 mice. Infect Immun. 2011;79:3861–71.

  21. 21.

    Fox JG, Rogers AB, Whary MT, Ge Z, Ohtani M, Jones EK, et al. Accelerated progression of gastritis to dysplasia in the pyloric antrum of TFF2 -/- C57BL6 x Sv129 Helicobacter pylori-infected mice. Am J Pathol. 2007;171:1520–8.

  22. 22.

    Rogers AB. Histologic scoring of gastritis and gastric cancer in mouse models. Methods Mol Biol. 2012;921:189–203.

  23. 23.

    Sung JaeKyu. Diagnosis and management of gastric dysplasia. Korean J Intern Med. 2016;31:201–9.

  24. 24.

    Ruchaud-Sparagano MH, Westley BR, May FE. The trefoil protein TFF1 is bound to MUC5AC in human gastric mucosa. Cell Mol Life Sci. 2004;61:1946–54.

  25. 25.

    Matsuda K, Yamauchi K, Matsumoto T, Sano K, Yamaoka Y, Ota H. Quantitative analysis of the effect of Helicobacter pylori on th.e expressions of SOX2, CDX2, MUC2, MUC5AC, MUC6, TFF1, TFF2, and TFF3 mRNAs in human gastric carcinoma cells. Scand J Gastroenterol. 2008;43:25–33.

  26. 26.

    Taupin D, Podolsky DK. Trefoil factors: initiators of mucosal healing. Nat Rev Mol Cell Biol. 2003;4:721–32.

  27. 27.

    Tomasetto C, Masson R, Linares JL, Wendling C, Lefebvre O, Chenard MP, et al. pS2/TFF1 interacts directly with the VWFC cysteine-rich domains of mucins. Gastroenterology. 2000;118:70–80.

  28. 28.

    Clyne M, Dillon P, Daly S, O’Kennedy R, May FE, Westley BR, et al. Helicobacter pylori interacts with the human single-domain trefoil protein TFF1. Proc Natl Acad Sci USA. 2004;101:7409–14.

  29. 29.

    Lefebvre O, Chenard MP, Masson R, Linares J, Dierich A, LeMeur M, et al. Gastric mucosa abnormalities and tumorigenesis in mice lacking the pS2 trefoil protein. Science. 1996;274:259–62.

  30. 30.

    Christopoulos C, Papavassiliou E. Gastric neuroendocrine tumors: biology and management. Annals of Gastroenterology. 2005;18:127–40.

  31. 31.

    Choi E, Hendley AM, Bailey JM, Leach SD, Goldenring JR. Expression of activated Ras in gastric chief cells of mice leads to the full spectrum of metaplastic lineage transitions. Gastroenterology. 2016;150:918–30 e913.

  32. 32.

    Xia HH, Yang Y, Lam SK, et al. Aberrant epithelial expression of trefoil family factor 2 and mucin 6 in Helicobacter pylori infected gastric antrum, incisura, and body and its association with antralisation. J Clin Pathol. 2004;57:861–6.

  33. 33.

    Karasawa F, Shiota A, Goso Y, Kobayashi M, Sato Y, Masumoto J, et al. Essential role of gastric gland mucin in preventing gastric cancer in mice. J Clin Investig. 2012;122:923–34.

  34. 34.

    Pezhouh MK, Park JY. Gastric pyloric gland adenoma. Arch Pathol Lab Med. 2015;139:823–6.

  35. 35.

    Markowski AR, Markowska A, Guzinska-Ustymowicz K. Pathophysiological and clinical aspects of gastric hyperplastic polyps. World J Gastroenterol. 2016;22:8883–91.

  36. 36.

    Ng GZ, Menheniott TR, Every AL, Stent A, Judd LM, Chionh YT, et al. The MUC1 mucin protects against Helicobacter pylori pathogenesis in mice by regulation of the NLRP3 inflammasome. Gut. 2016;65:1087–99.

  37. 37.

    Hoffmann W. TFF2, a MUC6-binding lectin stabilizing the gastric mucus barrier and more (Review). Int J Oncol. 2015;47:806–16.

  38. 38.

    Thompson LJ, Danon SJ, Wilson JE, O’Rourke JL, Salama NR, Falkow S, et al. Chronic Helicobacter pylori infection with Sydney strain 1 and a newly identified mouse-adapted strain (Sydney strain 2000) in C57BL/6 and BALB/c mice. Infect Immun. 2004;72:4668–79.

  39. 39.

    Navabi N, Johansson ME, Raghavan S, Lindén SK. Helicobacter pylori infection impairs the mucin production rate and turnover in the murine gastric mucosa. Infect Immun. 2013;81:829–37.

  40. 40.

    Guang W, Ding H, Czinn SJ, et al. Muc1 cell surface mucin attenuates epithelial inflammation in response to a common mucosal pathogen. J Biol Chem. 2010;285:20547–57.

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The authors thank the histology core personnel of the Division of Comparative Medicine for their technical support in the tissue processing and standardization of various immunohistochemical protocols. We also thank NIEHS histology core for their support.


This study is supported by NIH T32-OD010978, P01-CA028842, P30-ES002109 (to JGF) and R01-CA093405 (to TCW and JGF).

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Correspondence to James G. Fox.

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Muthupalani, S., Ge, Z., Joy, J. et al. Muc5ac null mice are predisposed to spontaneous gastric antro-pyloric hyperplasia and adenomas coupled with attenuated H. pylori-induced corpus mucous metaplasia. Lab Invest (2019) doi:10.1038/s41374-019-0293-y

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