Aberrant intestinal expression and allelic variants of mucin genes associated with inflammatory bowel disease

Loss of intestinal mucosa integrity is an important factor in the pathogenesis of inflammatory bowel disease (IBD). The aim of this study was to characterize expression changes and allelic variants of genes related to intestinal epithelial barrier function in this disease. Therefore, ileal and colonic mucosal biopsies from nonaffected regions of patients with ulcerative colitis (UC) and Crohn’s disease (CD), as well as non-IBD probands, were subjected to Affymetrix DNA-microarray analysis. Real-time reverse transcription polymerase chain reaction was used for verification in larger IBD sample numbers. Disturbed mRNA expression was identified for several mucin genes in both disease groups and tissues. A significant downregulation in the colon was obtained for MUC2 in CD and MUC12 in CD and UC. Expression analysis of all dysregulated mucins in a broad human tissue panel revealed dominant epithelial tissue-specific transcription. In silico analysis of the regulatory regions of these mucins indicated nuclear factor κB (NFκB) binding sites in each promoter. Furthermore, NFκB was overrepresented in mucin promoters and a component of a specific combination of transcription factors (composite module). In vivo stimulation experiments in the adenocarcinoma cell line LS174T showed inducible mucin expression by the cytokines tumor necrosis factor-α and transforming growth factor-β, which could be blocked by NFκB signaling inhibitors. Allelic discrimination screening obtained statistically significant associations for the MUC2–V116M (P = 0.003) polymorphism with CD and for MUC4–A585S (P = 0.025), as well as MUC13–R502S (P = 0.0003) with UC. These data suggest that the disturbed expression of mucin genes and the connection to the NFκB pathway may influence the integrity of the intestine and therefore contribute to the pathophysiology of IBD.

[1]  Alexander E. Kel,et al.  TRANSFAC®: transcriptional regulation, from patterns to profiles , 2003, Nucleic Acids Res..

[2]  D. Swallow,et al.  Molecular cloning of cDNAs derived from a novel human intestinal mucin gene. , 1990, Biochemical and biophysical research communications.

[3]  D. Swallow,et al.  Human mucin genes assigned to 11p15.5: identification and organization of a cluster of genes. , 1996, Genomics.

[4]  Alexander E. Kel,et al.  MATCHTM: a tool for searching transcription factor binding sites in DNA sequences , 2003, Nucleic Acids Res..

[5]  J. van Marle,et al.  MUC5B is the prominent mucin in human gallbladder and is also expressed in a subset of colonic goblet cells. , 1998, The American journal of physiology.

[6]  S. Targan,et al.  Tumor necrosis factor: biology and therapeutic inhibitors. , 2000, Gastroenterology.

[7]  Jason C. Mills,et al.  DNA microarrays and beyond: completing the journey from tissue to cell , 2001, Nature Cell Biology.

[8]  Y. Kim,et al.  Molecular cloning of human intestinal mucin cDNAs. Sequence analysis and evidence for genetic polymorphism. , 1989, The Journal of biological chemistry.

[9]  Judy H. Cho,et al.  Functional annotation of a novel NFKB1 promoter polymorphism that increases risk for ulcerative colitis. , 2004, Human molecular genetics.

[10]  T. Shuto,et al.  Transforming Growth Factor-β-Smad Signaling Pathway Cooperates with NF-κB to Mediate NontypeableHaemophilus influenzae-induced MUC2 Mucin Transcription* , 2002, The Journal of Biological Chemistry.

[11]  E. Cario.  BACTERIAL INTERACTIONS WITH CELLS OF THE INTESTINAL MUCOSA: TOLL-LIKE RECEPTORS AND NOD2 , 2005, Gut.

[12]  D. Podolsky,et al.  Inflammatory bowel disease. , 2002, The New England journal of medicine.

[13]  I. van Seuningen,et al.  Characterization of human mucin gene MUC4 promoter: importance of growth factors and proinflammatory cytokines for its regulation in pancreatic cancer cells. , 2001, The Journal of biological chemistry.

[14]  J. Rossen,et al.  The MUC family: an obituary. , 2002, Trends in biochemical sciences.

[15]  L. McIntyre,et al.  Combining mapping and arraying: An approach to candidate gene identification , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[16]  Reen Wu,et al.  Genome-wide search and identification of a novel gel-forming mucin MUC19/Muc19 in glandular tissues. , 2004, American journal of respiratory cell and molecular biology.

[17]  Judy H. Cho,et al.  [Letters to Nature] , 1975, Nature.

[18]  E. Wingender,et al.  MATCH: A tool for searching transcription factor binding sites in DNA sequences. , 2003, Nucleic acids research.

[19]  S. Brant,et al.  Inflammatory Bowel Disease Gene Hunting by Linkage Analysis: Rationale, Methodology, and Present Status of the Field , 2004, Inflammatory bowel diseases.

[20]  J. Davies,et al.  MUC5AC, but not MUC2, is a prominent mucin in respiratory secretions , 1996, Glycoconjugate Journal.

[21]  Edgar Wingender,et al.  A Novel Computational Approach for the Prediction of Networked Transcription Factors of Aryl Hydrocarbon-Receptor-Regulated Genes , 2004, Molecular Pharmacology.

[22]  R. Walker,et al.  Sulphation of colonic and rectal mucin in inflammatory bowel disease: reduced sulphation of rectal mucus in ulcerative colitis. , 1992, Clinical science.

[23]  J. Dekker,et al.  63 DISEASE‐RELATED QUANTITATIVE ALTERATIONS IN MUC2 SYNTHESIS IN ULCERATIVE COLITIS ARE REGULATED AT THE TRANSLATIONAL LEVEL , 1996, Journal of Pediatric Gastroenterology and Nutrition.

[24]  Chang-Hoon Kim,et al.  Extracellular signal-regulated kinase is involved in tumor necrosis factor-α-induced MUC5AC gene expression in cultured human nasal polyp epithelial cells , 2004, Acta oto-laryngologica.

[25]  A. Young,et al.  Prokaryotic regulation of epithelial responses by inhibition of IkappaB-alpha ubiquitination. , 2000, Science.

[26]  Y. Nakamura,et al.  Association of ulcerative colitis with rare VNTR alleles of the human intestinal mucin gene, MUC3. , 1999, Human molecular genetics.

[27]  P. Desreumaux,et al.  Abnormalities in mucin gene expression in Crohn's disease , 1999 .

[28]  T. Abe,et al.  mRNA of MUC2 is stimulated by IL‐4, IL‐13 or TNF‐α through a mitogen‐activated protein kinase pathway in human colon cancer cells , 2003, Immunology and cell biology.

[29]  I. van Seuningen,et al.  Sequence of the 5'-flanking region and promoter activity of the human mucin gene MUC5B in different phenotypes of colon cancer cells. , 2000, The Biochemical journal.

[30]  S. Batra,et al.  Cloning and sequencing of a human pancreatic tumor mucin cDNA. , 1990, The Journal of biological chemistry.

[31]  R. Newcombe,et al.  Thickness of adherent mucus gel on colonic mucosa in humans and its relevance to colitis. , 1994, Gut.

[32]  J. Dekker,et al.  MUC2 is the prominent colonic mucin expressed in ulcerative colitis. , 1996, Gut.

[33]  S. Markowitz,et al.  Loss of transforming growth factor β signalling in the intestine contributes to tissue injury in inflammatory bowel disease , 2001, Gut.

[34]  Mourad Sahbatou,et al.  Association of NOD2 leucine-rich repeat variants with susceptibility to Crohn's disease , 2001, Nature.

[35]  Gerd Schmitz,et al.  Loss of detoxification in inflammatory bowel disease: dysregulation of pregnane X receptor target genes. , 2004, Gastroenterology.

[36]  A. Young,et al.  Prokaryotic Regulation of Epithelial Responses by Inhibition of IκB-α Ubiquitination , 2000 .

[37]  V. G. Dupérat,et al.  Structural features of the core proteins of human airway mucins ascertained by cDNA cloning. , 1991, The American review of respiratory disease.

[38]  Kan Yang,et al.  Colorectal Cancer in Mice Genetically Deficient in the Mucin Muc2 , 2002, Science.

[39]  R. V. D. van der Hulst,et al.  H pylori colocalises with MUC5AC in the human stomach , 2000, Gut.

[40]  C. Probert,et al.  Mucins and inflammatory bowel disease , 2000, Postgraduate medical journal.

[41]  G. Lathrop,et al.  Associations of distinct variants of the intestinal mucin gene MUC3A with ulcerative colitis and Crohn's disease , 2001, Journal of Human Genetics.

[42]  J. Terwilliger,et al.  Two stage genome–wide search in inflammatory bowel disease provides evidence for susceptibility loci on chromosomes 3, 7 and 12 , 1996, Nature Genetics.

[43]  J. Rhodes,et al.  Increased rate of sialylation of colonic mucin by cultured ulcerative colitis mucosal explants. , 1995, Digestion.

[44]  Sankar Ghosh,et al.  Signaling to NF-kappaB. , 2004, Genes & development.

[45]  B. Aggarwal,et al.  Caffeic acid phenethyl ester is a potent and specific inhibitor of activation of nuclear transcription factor NF-kappa B. , 1996, Proceedings of the National Academy of Sciences of the United States of America.