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The role of the NOD2 gene in the pathogenesis of Crohn’s disease

By T.A. Chalmers-Watson


Introduction: Crohn’s disease is characterised by an abnormal inflammatory response possibly induced by components of enteric bacteria, in genetically susceptible individuals. Mutations in the NOD2 gene are strongly associated with Crohn's disease, although the mechanisms by which these mutations cause Crohn’s disease remain unknown. Peripheral blood monocytes (PBMC), a key component of the innate immune system, highly express NOD2, as do intestinal epithelial Paneth cells. The ligand for NOD2 is Muramyl dipeptide (MDP), a component of bacterial peptidoglycan. MDP has been shown to be a powerful priming agent for subsequent stimulation by lipopolysaccharide (LPS) in cell lines and this effect has been linked to NOD2. Studies of primary mononuclear cells in Crohn’s disease comparing the functional effects of the NOD2 mutations had not previously been reported. Aims: To determine the effect of inherited mutations in the NOD2 gene on the cellular responses of freshly extracted PBMC to MDP and other bacterial ligands including mycobacteria. To examine the effect of MDP pre treatment ‘priming’ on PBMC responses to subsequent LPS stimulation in both normal and Crohn’s disease affected patients expressing wild type and mutant NOD2 proteins. Methods: PBMC from healthy controls (n=12), and Crohn’s disease affected patients who were genotypically either wild type (n=12), heterozygous (n=11) or homozygous (n=5) for the common disease-causing NOD2 mutations. PBMC were stimulated with bacterial products in vitro, with or without prior stimulation or ‘priming’ with MDP. The transcription of selected cytokine genes was determined by real time quantitative RT-PCR Results: MDP is a weak stimulant of inflammatory responses in PBMC whereas LPS evoked much stronger responses. Responses to MDP were particularly reduced in PBMC homozygous for the NOD2 mutations. Priming with MDP reduced the inflammatory response of normal PBMC to subsequent LPS stimulation. In PBMC carrying two mutant NOD2 alleles this modulatory effect was reversed and MDP priming caused the inflammatory response to be enhanced. Conclusion: MDP priming significantly modulates responses of monocytes to LPS. This effect is altered in patients with Crohn’s disease – possibly related to mutations in the NOD2 gene. This modulatory effect may explain in part the pro-inflammatory consequence of mutations in the NOD2 gene and could provide mechanistic understanding of how mutations in the NOD2 gene may cause Crohn’s disease

Publisher: UCL (University College London)
Year: 2009
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Provided by: UCL Discovery

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  1. (2003). (a) ‘Lessons from Nod2 studies: towards a link between Crohn's disease and bacterial sensing’ Trends Immunol.
  2. 2002‘The origin of the synergistic effect of muramyl dipeptide with endotoxin and peptidoglycan’
  3. (2001). 2005(a)‘Synergy between TLR9 and NOD2 innate immune responses is lost in genetic Crohn's disease.’ Gut.
  4. (2007). A brief history of T(H)17, the first major revision in the T(H)1/T(H)2 hypothesis of T cell-mediated tissue damage’. Nat Med. Feb;13(2):139-45 172. Stockton JC,
  5. (2006). A genome-wide association study identifies IL23R as an inflammatory bowel disease gene’
  6. (1999). A genome-wide search identifies potential new susceptibility loci for Crohn's disease", Inflamm.Bowel.Dis.5:4: 271-278 115. Maeda
  7. (2007). A nonsynonymous SNP in ATG16L1 predisposes to ileal Crohn's disease and is independent of CARD15 and IBD5.’ Gastroenterology.
  8. (2007). A population-based case-control study of CARD15 and other risk factors in Crohn's disease and ulcerative colitis’
  9. (1998). American families with Crohn's disease have strong evidence for linkage to chromosome 16 but not chromosome 12",
  10. (2001). Association of NOD2 leucine-rich repeat variants with susceptibility to Crohn's disease’
  11. (2001). Autoimmunity and apoptosis: the Crohn's connection’ Immunity. 15:5-14. 129 15. Bevins C L.
  12. (2001). Behaviour of Crohn's disease according to the Vienna classification: changing pattern over the course of the disease.’
  13. (2004). CARD15 gene mutations in periodontitis" J.Clin.Periodonto 31:10:
  14. (2001). CARD15 mutations in Blau syndrome",
  15. (2003). CARD15/NOD2 gene variants are associated with familially occurring and complicated forms of Crohn's disease."
  16. (2002). CARD15/NOD2 mutational analysis and genotype-phenotype correlation in 612 patients with inflammatory bowel disease.’
  17. (2005). CATERPILLER: a novel gene family important in immunity, cell death, and diseases’
  18. (2002). Clinical aspects and pathophysiology of inflammatory bowel disease’
  19. (2004). Clinical epidemiology of inflammatory bowel disease: Incidence, prevalence, and environmental influences’
  20. (1998). Complementation cloning of NEMO, a component of the IkappaB kinase complex essential for NF-kappaB activation",
  21. (1998). Concordance rates of twins and siblings in inflammatory bowel disease.’ Gut ; 42:A40 174.
  22. (2003). Crohn's disease and the NOD2 gene: a role for paneth cells.’
  23. (1990). Crohn's disease in the city of Derby,
  24. (2003). Crohn's disease-associated NOD2 variants share a signalling defect in response to lipopolysaccharide and peptidoglycan‘
  25. (2003). Crohn's disease: the cold chain hypothesis’ Lancet;362(9400):2012-5 86. Hugot
  26. (2001). Dealing the CARDs between life and death’
  27. (2003). Defining complex contributions of NOD2/CARD15 gene mutations, age at onset, and tobacco use on Crohn's disease phenotypes",
  28. (2004). Differential effects of NOD2 variants on Crohn's disease risk and phenotype in diverse populations: a metaanalysis’
  29. (2005). Does Mycobacterium avium subspecies paratuberculosis cause Crohn's disease’
  30. (2005). Early-onset sarcoidosis and CARD15 mutations with constitutive nuclear factor-kappaB activation: common genetic etiology with Blau syndrome’ Blood. 1;105(3):1195-7 96. Keshav
  31. (2003). Effects of nonpathogenic bacteria on cytokine secretion by human intestinal mucosa’
  32. (2004). Enhanced Escherichia coli adherence and invasion in Crohn's disease and colon
  33. (1987). Enhancement of endotoxin lethality and generation of anaphylactoid reactions by lipopolysaccharides in muramyl-dipeptide treated mice.’ Infect Immun 55:409-413 176. Takeda
  34. (1994). Enteral nutrition as primary therapy in Crohn's disease’ Gut.
  35. (2008). Enteric salmonella infection inhibits Paneth cell antimicrobial peptide expression",
  36. (2003). Expression of NOD2 in Paneth cells: a possible link to Crohn's ileitis’
  37. (1991). Familial occurrence of inflammatory bowel disease",
  38. (2002). Forbes A et al
  39. (2006). Genetic advances in inflammatory bowel disease’ Curr Treat Options Gastroenterol.;9(3):191-200.
  40. (1999). Genetic analysis in Italian families with inflammatory bowel disease supports linkage to the IBD1 locus--a GISC study’.
  41. (1998). Genetic epidemiology in inflammatory bowel disease",
  42. (2003). Genetic variation and activity of mouse Nod2, a susceptibility gene for Crohn's disease’.
  43. (2007). Genome-wide association study identifies new susceptibility loci for Crohn’s disease and implicates autophagy in disease pathogenesis’
  44. Gordon JI.2003 ‘Angiogenins: a new class of microbicidal proteins involved in innate immunity’.
  45. (2005). GRIM-19 interacts with nucleotide oligomerization domain 2 and serves as downstream effector of anti-bacterial function in intestinal epithelial cells’
  46. (2004). High prevalence of adherent-invasive Escherichia coli associated with ileal mucosa in Crohn's disease.’ Gastroenterology. 127(2):412-21 46. Davis RL, Bohike K.
  47. (2005). High prevalence of Mycobacterium avium subspecies paratuberculosis IS900 DNA in gut tissues from individuals with Crohn's disease’
  48. (2000). High-density genome scan in Crohn’s disease shows confirmed linkage to chromosome 14q11-12",
  49. (2003). Host recognition of bacterial muramyl dipeptide mediated through NOD2. Implications for Crohn's disease.’ J.Biol.Chem ;278:5509-5512 93. Inohara
  50. (1995). How effective is enteral nutrition in inducing clinical remission in active Crohn's disease? A meta-analysis of the randomized clinical trials’
  51. (1999). Human CARD4 protein is a novel CEDApaf-1 cell death family member that activates NF-kappaB’,
  52. (2001). Human defensin 5 is stored in precursor form in normal Paneth cells and is expressed by some villous epithelial cells and by metaplastic Paneth cells in the colon in inflammatory bowel disease",
  53. (2001). Human Nod1 confers responsiveness to bacterial lipopolysaccharides.’
  54. (2004). Identification of bacterial muramyl dipeptide as activator of the NALP3/cryopyrin inflammasome’
  55. (1998). Identification of novel susceptibility loci for inflammatory bowel disease on chromosomes 1p, 3q, and 4q: evidence for epistasis between 1p and IBD1",
  56. (2000). Immune potentiation of ultrafine dietary particles in normal subjects and patients with inflammatory bowel disease’
  57. (1999). Impaired on/off regulation of TNF biosynthesis in mice lacking TNF AU-rich elements: implications for joint and gutassociated immunopathologies",
  58. (2002). In siblings with similar genetic susceptibility for inflammatory bowel disease, smokers tend to develop Crohn's disease and non-smokers develop ulcerative colitis’ Gut. 51(1):21-5 28. Cadwell
  59. (2003). Increased expression of antimicrobial peptides and lysozyme in colonic epithelial cells of patients with ulcerative colitis",
  60. (2003). Increased inflammation in lysozyme M-deficient mice in response to Micrococcus luteus and its peptidoglycan.’ Blood. 101(6):2388-92 66.
  61. (1994). Increased risk of inflammatory bowel disease associated with oral contraceptive use’
  62. (2002). Induction of Nod2 in myelomonocytic and intestinal epithelial cells via nuclear factor-kappa B activation’,
  63. (1995). Inflammatory bowel disease and adenomas in mice expressing a dominant negative N-cadherin."
  64. (2000). Inflammatory bowel disease: epidemiology and management in an English general practice population’ Aliment Pharmacol Ther.14
  65. (1998). Inflammatory bowel disease: etiology and pathogenesis’.
  66. (2006). Interleukin-23 drives innate and T cell-mediated intestinal inflammation’ J Exp Med.
  67. (1990). Interleukin-8 gene expression by a p ulmonary epithelial cell line. A model for cytokine networks in the lung’
  68. (2001). International collaboration provides convincing linkage replication in complex disease through analysis of a large pooled data set: Crohn disease and chromosome 16’
  69. (1993). Is small intestinal permeability really increased in relatives of patients with Crohn's disease?’ Gastroenterology.
  70. (1996). IS900 PCR to detect Mycobacterium paratuberculosis in retail supplies of whole pasteurized cows'
  71. (1998). Lederman E et al
  72. (1998). Linkage and association between inflammatory bowel disease and a locus on chromosome 12",
  73. (1992). Lysozyme gene expression in inflammatory bowel disease",
  74. (2001). Measles virus and Crohn's disease: a critical appraisal of the current literature’ Inflamm Bowel Dis.7(1):51-7 156. Rosenstiel
  75. (1997). Micrococcus luteus cells and cell walls induce anaphylactoid reactions accompanied by early death and serum cytokines in mice primed with muramyl dipeptide’ FEMS Immunol Med Microbiol.
  76. (2005). Muramyl dipeptide and toll-like receptor sensitivity in NOD2-associated Crohn's disease’ Lancet.
  77. (2004). Muramyldipeptide and diaminopimelic acid –containing desmuramylpeptides in combination with chemically synthesised Toll like receptor agonists synergistically induced production of interleukin 8 in a NOD2 and NOD1 dependent manner, respectively,
  78. (2002). NF-kappaB regulation in the immune system",
  79. (2004). NOD2 (CARD15) mutations in Crohn's disease are associated with diminished mucosal alpha-defensin expression"
  80. (2005). NOD2 and toll-like receptors are nonredundant recognition systems of Mycobacterium tuberculosis’
  81. (2004). NOD2 is a negative regulator of Toll-like receptor 2-mediated T helper type 1 responses’.
  82. (2005). Nod2-dependent regulation of innate and adaptive immunity in the intestinal tract",
  83. (2001). Nod2, a Nod1/Apaf-1 family member that is restricted to monocytes and activates NF-kappaB’ J.Biol.Chem 276:
  84. (2006). NOD2: ethnic and geographic differences’
  85. (2003). NOD2/CARD15 gene mutation is not associated with susceptibility to Wegener's granulomatosis."
  86. (2005). NOD2/CARD15 gene polymorphisms in Crohn's disease: a genotype-phenotype analysis in Danish and Portuguese patients and controls’ Digestion.
  87. (2002). Nods: a family of cytosolic proteins that regulate the host response to pathogens’ Curr.Opin.Microbiol.
  88. (2005). Paneth cells: their role in innate immunity and inflammatory disease.’ Gut 54(12):1802-9 54. Fabia R,
  89. (2003). Pathogenic Yersinia DNA is detected in bowel and mesenteric lymph nodes from patients with Crohn's disease’
  90. (2003). Pharmacokinetics and feeding responses to muramyl dipeptide in rats’ Physiol Behav.
  91. (2001). Plant pathogens and integrated defence responses to infection",
  92. (2004). Recognition of commensal microflora by toll-like receptors is required for intestinal homeostasis’ Cell;118(2):229-41 138 150. Reinecker
  93. (2004). Regulation of IL-8 and IL-1beta expression in Crohn's disease associated NOD2/CARD15 mutations"
  94. (1999). Regulation of intestinal alpha-defensin activation by the metalloproteinase matrilysin in innate host defense",
  95. (2003). Role of nod2 in the response of macrophages to toll-like receptor agonists",
  96. (1997). Rutgeerts P for The Crohn's Disease
  97. Sansonetti PJ.2004 ‘War and peace at mucosal surfaces’ Nat Rev Immunol.
  98. (2004). Serum angiogenin in inflammatory bowel disease.’ Dig Dis Sci.
  99. (2001). Smoking cessation and the course of Crohn's disease: an intervention
  100. (1994). Smoking habits and recurrence in Crohn's disease’ Gastroenterology;106(3):643-8 39.
  101. (2007). SOX9 is required for the differentiation of paneth cells in the intestinal epithelium.’ Gastroenterology. 133(2):539-46 129. Motiwala
  102. (1996). Susceptibility locus for inflammatory bowel disease on chromosome 16 has a role in Crohn's disease, but not in ulcerative colitis",
  103. (2001). TAK1 is a ubiquitin-dependent kinase of
  104. (2004). The 3020insC mutation of the NOD2/CARD15 gene in patients with periodontal disease"
  105. (1997). The CARD domain: a new apoptotic signalling motif",
  106. (2002). The contribution of NOD2 131 gene mutations to the risk and site of disease in inflammatory bowel disease’
  107. (2004). The Crohn's disease protein, NOD2, requires RIP2 in order to induce ubiquitinylation of a novel site on NEMO’ Curr Biol;14(24):2217-27 2. Abraham
  108. (2004). The Gastrointestinal System at a Glance’. Chapter 34. Ulcerative Colitis and Crohn's disease.
  109. (2002). The genetics of inflammatory bowel disease’ Gastroenterology.
  110. (1994). The germfree state prevents development of gut and joint inflammatory disease in HLA-B27 transgenic rats’
  111. (2003). The immunological and genetic basis of inflammatory bowel disease‘
  112. (1997). The influence of normal microbial flora on the development of chronic mucosal inflammation",
  113. (2001). The NOD: a signaling module that regulates apoptosis and host defense against pathogens.’
  114. (2004). The role of Toll-like receptors and Nod proteins in bacterial infection’
  115. (1999). The role of tumor necrosis factor in health and disease",
  116. (1996). Two stage genome-wide search in inflammatory bowel disease provides evidence for susceptibility loci on chromosomes 3, 7 and 12",
  117. (1997). Two-year-outcomes analysis of Crohn's disease treated with rifabutin and macrolide antibiotics.’
  118. (1988). Ulcerative colitis and Crohn's disease in an unselected population of monozygotic and dizygotic twins. A study of heritability and the influence of smoking.’
  119. (2002). Using COX-2 inhibitors in IBD: anti-inflammatories inflame a controversy’
  120. (2003). Uveitis in patients with sarcoidosis is not associated with mutations in NOD2 (CARD15)"

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