title: TLE1 modifies the effects of NOD2 in the pathogenesis of Crohn's disease.
abstract: OBJECTIVE
  The mechanisms by which specific mutations in NOD2/CARD15 increase the risk for Crohn's disease (CD) are unclear. We identified proteins that interact with NOD2 and investigated them by expression, genetic, and functional analyses.
  
  
  METHODS
  By using a yeast 2-hybrid screen of an intestinal epithelial library, we identified proteins that interact with NOD2 and confirmed the interactions in mammalian cells using co-immunoprecipitation. We used microarray analysis to analyze gene expression patterns in 302 intestinal biopsy samples (129 from patients with ulcerative colitis [UC], 106 with CD, and 67 controls). Eighty single-nucleotide polymorphisms within the genes that encoded 6 interacting proteins were genotyped in a discovery cohort (869 cases of inflammatory bowel disease [IBD], 885 controls) and a replication cohort (504 patients with IBD, 713 controls). We investigated interaction between transducin-like enhancer of split 1 (TLE1) and NOD2 in HEK293 cells.
  
  
  RESULTS
  We identified 6 NOD2-interacting proteins (TLE1, UDP-N-acetyl-alpha-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase 2 [GALNT2], HIV-1 Tat interactive protein [HTATIP], Vimentin, fission 1 (mitochondrial outer membrane) homolog [FIS1], and protein phosphatase 2, regulatory subunit B', epsilon isoform [PPP2R5E]). Of these, expression of GALNT2 (CD, P = .004) and vimentin (CD, P = .006; UC, P = .0025) was altered in patients with IBD compared with controls. Single-nucleotide polymorphisms within TLE1 were associated with susceptibility to CD, specifically with ileal disease (rs6559629, P = 3.1 × 10⁻⁵; odds ratio, 1.45). The TLE1 risk allele is required for susceptibility to CD in carriers of NOD2 mutations. In cells, TLE1 and NOD2 co-localized around the nuclear membrane and TLE1 inhibited activation of nuclear factor-κB by NOD2.
  
  
  CONCLUSIONS
  Epistatic and biological interactions between TLE1 and NOD2 are involved in IBD pathogenesis. NOD2 might be involved in a series of pathways such as epigenetic regulation of expression (via TLE1 and HTATIP), biosynthesis of mucin (via GALNT2), apoptosis (via PPP2R5E and FIS1), and integrity of the intracellular cytoskeleton (vimentin).
authors.affiliation: Gastrointestinal Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh, Molecular Medicine Centre, Western General Hospital, Edinburgh, UK. enimmo@ed.ac.uk
authors.forename: Elaine R
authors.initials: ER
authors.lastname: Nimmo
authors.forename: Craig
authors.initials: C
authors.lastname: Stevens
authors.forename: Anne M
authors.initials: AM
authors.lastname: Phillips
authors.forename: Amanda
authors.initials: A
authors.lastname: Smith
authors.forename: Hazel E
authors.initials: HE
authors.lastname: Drummond
authors.forename: Colin L
authors.initials: CL
authors.lastname: Noble
authors.forename: Michael
authors.initials: M
authors.lastname: Quail
authors.forename: Gail
authors.initials: G
authors.lastname: Davies
authors.forename: Marian C
authors.initials: MC
authors.lastname: Aldhous
authors.forename: David C
authors.initials: DC
authors.lastname: Wilson
authors.forename: Jack
authors.initials: J
authors.lastname: Satsangi
mesh_terms: D001706:Biopsy; D016022:Case-Control Studies; D056970:Co-Repressor Proteins; D003093:Colitis, Ulcerative / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D003106:Colon / Q000378:metabolism / Q000473:pathology; D003424:Crohn Disease / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology*; D004843:Epistasis, Genetic / Q000502:physiology*; D051548:Histone Acetyltransferases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D000074443:Lysine Acetyltransferase 5; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D046228:Microarray Analysis; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D017350:N-Acetylgalactosaminyltransferases / Q000235:genetics / Q000378:metabolism; D053473:Nod2 Signaling Adaptor Protein / Q000235:genetics* / Q000502:physiology*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D054648:Protein Phosphatase 2 / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics* / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology; D014746:Vimentin / Q000235:genetics / Q000378:metabolism; D000097763:Polypeptide N-acetylgalactosaminyltransferase
chemical_list: D056970:Co-Repressor Proteins; D051548:Histone Acetyltransferases; D000074443:Lysine Acetyltransferase 5; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D017350:N-Acetylgalactosaminyltransferases; D053473:Nod2 Signaling Adaptor Protein; D054648:Protein Phosphatase 2; D012097:Repressor Proteins; D014746:Vimentin; D000097763:Polypeptide N-acetylgalactosaminyltransferase; C476525:FIS1 protein, human; C429018:NOD2 protein, human; C525692:PPP2R5E protein, human; C499043:TLE1 protein, human; C098489:KAT5 protein, human