NOD2 status and human ileal gene expression†‡

Background: NOD2 single nucleotide polymorphisms have been associated with increased risk of ileal Crohn's disease (CD). This exploratory study was conducted to compare ileal mucosal gene expression in CD patients with and without NOD2 risk alleles. Methods: Ileal samples were prospectively collected from 18 nonsmoking CD patients not treated with anti‐TNF‐&agr; biologics and 9 nonsmoking control patients without inflammatory bowel disease undergoing initial resection and genotyped for the 3 major NOD2 risk alleles (Arg702Trp, Gly908Arg, Leu1007fs). Microarray analysis was performed in samples from 4 NOD2R (at least 1 risk allele) CD patients, 4 NOD2NR (no risk alleles) CD patients, and 4 NOD2NR controls. Candidate genes selected by significance analysis of microarrays (SAM) were confirmed by quantitative reverse transcriptase polymerase chain reaction (qRT‐PCR) assays of all the samples. Results: SAM detected upregulation of 18 genes in affected ileum in NOD2R compared to NOD2NR CD patients, including genes related to lymphocyte activation. SAM also detected altered ileal gene expression in unaffected NOD2NR ileal mucosal CD samples compared to NOD2NR control samples. qRT‐PCR conducted on all the samples confirmed that increased CD3D expression in affected samples was associated with NOD2R status, and that increased MUC1, DUOX2, DMBT1 and decreased C4orf7 expression in unaffected samples was associated with CD, independent of NOD2 status. Conclusions: The results support the concept that NOD2 risk alleles contribute to impaired regulation of inflammation in the ileum. Furthermore, altered ileal gene expression, independent of NOD2 status, is detected in the unaffected proximal margin of resected ileum from CD patients. (Inflamm Bowel Dis 2010)

[1]  J. Ioannidis,et al.  Differential Effects of NOD2 Variants on Crohn's Disease Risk and Phenotype in Diverse Populations: A Metaanalysis , 2004, The American Journal of Gastroenterology.

[2]  S. Davies,et al.  Crohn's disease and the NOD2 gene: a role for paneth cells. , 2003, Gastroenterology.

[3]  L. Simms,et al.  Reduced α-defensin expression is associated with inflammation and not NOD2 mutation status in ileal Crohn’s disease , 2008, Gut.

[4]  A. Sharpe,et al.  CD48 controls T-cell and antigen-presenting cell functions in experimental colitis. , 2006, Gastroenterology.

[5]  E. Clark,et al.  FDC-SP, a Novel Secreted Protein Expressed by Follicular Dendritic Cells1 , 2002, The Journal of Immunology.

[6]  A. Sepulveda,et al.  Cutting Edge: Transgenic Expression of Human MUC1 in IL-10−/− Mice Accelerates Inflammatory Bowel Disease and Progression to Colon Cancer1 , 2007, The Journal of Immunology.

[7]  Y. Natkunam,et al.  Jaw1/LRMP, a germinal centre‐associated marker for the immunohistological study of B‐cell lymphomas , 2006, The Journal of pathology.

[8]  M. Cooper,et al.  Fc receptor-like proteins (FCRL): immunomodulators of B cell function. , 2007, Advances in experimental medicine and biology.

[9]  J. Bidart,et al.  Dual oxidase2 is expressed all along the digestive tract. , 2005, American journal of physiology. Gastrointestinal and liver physiology.

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

[11]  S. Collins,et al.  Structural abnormalities of the nervous system in Crohn's disease and ulcerative colitis , 1998, Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society.

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

[13]  M Schwab,et al.  NOD2 (CARD15) mutations in Crohn’s disease are associated with diminished mucosal α-defensin expression , 2004, Gut.

[14]  G. Parmigiani,et al.  Genome‐wide gene expression differences in Crohn's disease and ulcerative colitis from endoscopic pinch biopsies: Insights into distinctive pathogenesis , 2007, Inflammatory bowel diseases.

[15]  Judy H. Cho,et al.  Genome-wide association defines more than 30 distinct susceptibility loci for Crohn's disease , 2008, Nature Genetics.

[16]  Judy H. Cho,et al.  Chronic stimulation of Nod2 mediates tolerance to bacterial products , 2007, Proceedings of the National Academy of Sciences.

[17]  V. Korolik,et al.  MUC1 cell surface mucin is a critical element of the mucosal barrier to infection. , 2007, The Journal of clinical investigation.

[18]  J. Satsangi,et al.  The Montreal classification of inflammatory bowel disease: controversies, consensus, and implications , 2006, Gut.

[19]  Tomohiro Watanabe,et al.  The Molecular Basis of NOD2 Susceptibility Mutations in Crohn's Disease , 2008, Mucosal Immunology.

[20]  H. Drummond,et al.  Does Cigarette Smoking Influence the Phenotype of Crohn's Disease? Analysis Using the Montreal Classification , 2007, The American Journal of Gastroenterology.

[21]  M. Hermiston,et al.  CD45: a critical regulator of signaling thresholds in immune cells. , 2003, Annual review of immunology.

[22]  A. Fischer,et al.  CD3 deficiencies , 2005, Current opinion in allergy and clinical immunology.

[23]  K. Becker,et al.  CYFIP2 is highly abundant in CD4+ cells from multiple sclerosis patients and is involved in T cell adhesion , 2004, European journal of immunology.

[24]  Yan Yan,et al.  Risk Factors for Surgical Recurrence after Ileocolic Resection of Crohn’s Disease , 2008, Diseases of the colon and rectum.

[25]  J. Söderholm,et al.  Peyer's Patches and M Cells as Potential Sites of the Inflammatory Onset in Crohn's Disease , 2006, Annals of the New York Academy of Sciences.

[26]  R. Farmer,et al.  Long-term follow-up of patients with Crohn's disease. Relationship between the clinical pattern and prognosis. , 1985, Gastroenterology.

[27]  G. Rogler,et al.  Subtractive screening reveals up‐regulation of NADPH oxidase expression in Crohn's disease intestinal macrophages , 2001, Clinical and experimental immunology.

[28]  O. Nielsen,et al.  CARD15 Status and Familial Predisposition for Crohn's Disease and Colonic Gene Expression , 2007, Digestive Diseases and Sciences.

[29]  A. Poustka,et al.  DMBT1 confers mucosal protection in vivo and a deletion variant is associated with Crohn's disease. , 2007, Gastroenterology.

[30]  N. Warner,et al.  Function of Nod‐like receptors in microbial recognition and host defense , 2009, Immunological reviews.

[31]  Judy H. Cho,et al.  Expression of NOD2 in Paneth cells: a possible link to Crohn’s ileitis , 2003, Gut.

[32]  Laurent Beaugerie,et al.  The second European evidence-based Consensus on the diagnosis and management of Crohn's disease: Definitions and diagnosis. , 2006, Journal of Crohn's & colitis.

[33]  K. Yarasheski,et al.  Alterations in thigh subcutaneous adipose tissue gene expression in protease inhibitor-based highly active antiretroviral therapy. , 2005, Metabolism: clinical and experimental.

[34]  S. Targan,et al.  Mutations in NOD2 are associated with fibrostenosing disease in patients with Crohn's disease. , 2002, Gastroenterology.

[35]  Satoru Takahashi,et al.  The transcriptional programme of antibody class switching involves the repressor Bach2 , 2004, Nature.

[36]  M. Roberts,et al.  Novel markers of the human follicle-associated epithelium identified by genomic profiling and microdissection. , 2005, Gastroenterology.

[37]  Judy H. Cho,et al.  Defining Complex Contributions of NOD2/CARD15 Gene Mutations, Age at Onset, and Tobacco Use On Crohn's Disease Phenotypes , 2003, Inflammatory bowel diseases.

[38]  P. Rutgeerts,et al.  Predictability of the postoperative course of Crohn's disease. , 1990, Gastroenterology.

[39]  P. Rutgeerts,et al.  Mucosal healing in inflammatory bowel disease: impossible ideal or therapeutic target? , 2007, Gut.

[40]  A. Lacy,et al.  Crohn's Disease Patients Carrying Nod2/CARD15 Gene Variants Have an Increased and Early Need for First Surgery due to Stricturing Disease and Higher Rate of Surgical Recurrence , 2005, Annals of surgery.

[41]  B F Warren,et al.  European evidence based consensus on the diagnosis and management of Crohn’s disease: definitions and diagnosis , 2006, Gut.

[42]  A. Poustka,et al.  Regulation of DMBT1 via NOD2 and TLR4 in Intestinal Epithelial Cells Modulates Bacterial Recognition and Invasion1 , 2007, The Journal of Immunology.

[43]  T. Ahmad,et al.  The molecular classification of the clinical manifestations of Crohn's disease. , 2002, Gastroenterology.

[44]  A. Marshall,et al.  Follicular Dendritic Cell Secreted Protein (FDC-SP) Regulates Germinal Center and Antibody Responses1 , 2007, The Journal of Immunology.

[45]  B. Vainer,et al.  Expression of the genes dualoxidase2, lipocalin 2 and regenerating islet-derived 1 alpha in Crohn's disease , 2007, Scandinavian journal of gastroenterology.

[46]  Jan G. M. Bolscher,et al.  Identification of the Bacteria-binding Peptide Domain on Salivary Agglutinin (gp-340/DMBT1), a Member of the Scavenger Receptor Cysteine-rich Superfamily* , 2002, The Journal of Biological Chemistry.

[47]  R. Tibshirani,et al.  Significance analysis of microarrays applied to the ionizing radiation response , 2001, Proceedings of the National Academy of Sciences of the United States of America.