ABSTRACT
Inflammatory bowel diseases are chronic gastrointestinal inflammatory disorders that affect millions of people worldwide. Genome-wide association studies have identified 200 inflammatory bowel disease-associated loci, but few have been conclusively resolved to specific functional variants. Here we report fine-mapping of 94 inflammatory bowel disease loci using high-density genotyping in 67,852 individuals. We pinpoint 18 associations to a single causal variant with greater than 95% certainty, and an additional 27 associations to a single variant with greater than 50% certainty. These 45 variants are significantly enriched for protein-coding changes (n = 13), direct disruption of transcription-factor binding sites (n = 3), and tissue-specific epigenetic marks (n = 10), with the last category showing enrichment in specific immune cells among associations stronger in Crohn's disease and in gut mucosa among associations stronger in ulcerative colitis. The results of this study suggest that high-resolution fine-mapping in large samples can convert many discoveries from genome-wide association studies into statistically convincing causal variants, providing a powerful substrate for experimental elucidation of disease mechanisms.
Subject(s)
Genetic Predisposition to Disease/genetics , Genetic Variation/genetics , Inflammatory Bowel Diseases/genetics , Quantitative Trait Loci/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Binding Sites , Chromatin/genetics , Colitis, Ulcerative/genetics , Crohn Disease/genetics , Epigenesis, Genetic/genetics , Female , Genome-Wide Association Study , Genotype , Humans , Linkage Disequilibrium/genetics , Male , Middle Aged , Smad3 Protein/genetics , Transcription Factors/metabolism , Young AdultABSTRACT
BACKGROUND & AIMS: A few rare monogenic primary immunodeficiencies (PIDs) are characterized by chronic intestinal inflammation that resembles Crohn's disease (CD). We investigated whether 23 genes associated with 10 of these monogenic disorders contain common, low-frequency, or rare variants that increase risk for CD. METHODS: Common and low frequency variants in 1 Mb loci centered on the candidate genes were analyzed using meta-data corresponding to genotypes of approximately 17,000 patients with CD or without CD (controls) in Europe. The contribution of rare variants was assessed by high-throughput sequencing of 4750 individuals, including 660 early-onset and/or familial cases among the 2390 patients with CD. Variants were expressed from vectors in SW480 or HeLa cells and functions of their products were analyzed in immunofluorescence, luciferase, immunoprecipitation, and immunoblot assays. RESULTS: We reproduced the association of the interleukin 10 locus with CD (P = .007), although none of the significantly associated variants modified the coding sequence of interleukin 10. We found XIAP to be significantly enriched for rare coding mutations in patients with CD vs controls (P = .02). We identified 4 previously unreported missense variants associated with CD. Variants in XIAP cause the PID X-linked lymphoproliferative disease type 2, yet none of the carriers of these variants had all the clinical features of X-linked lymphoproliferative disease type 2. Identified XIAP variants S123N, R233Q, and P257A were associated with an impaired activation of NOD2 signaling after muramyl dipeptide stimulation. CONCLUSIONS: In a systematic analysis of variants in 23 PID-associated genes, we confirmed the association of variants in XIAP with CD. Further screenings for CD-associated variants and analyses of their functions could increase our understanding of the relationship between PID-associated genes and CD pathogenesis.
Subject(s)
Crohn Disease/genetics , Immunologic Deficiency Syndromes/genetics , X-Linked Inhibitor of Apoptosis Protein/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Belgium , Cells, Cultured , Child , Child, Preschool , Crohn Disease/blood , Crohn Disease/immunology , Female , Fluorescent Antibody Technique , France , High-Throughput Nucleotide Sequencing , Humans , Immunologic Deficiency Syndromes/blood , Immunologic Deficiency Syndromes/immunology , Interleukin-10/genetics , Male , Middle Aged , Monocytes , Mutation, Missense , Nod2 Signaling Adaptor Protein/metabolism , Primary Cell Culture , Sequence Analysis, DNA , Signal Transduction/genetics , Young AdultABSTRACT
GWAS have identified >200 risk loci for Inflammatory Bowel Disease (IBD). The majority of disease associations are known to be driven by regulatory variants. To identify the putative causative genes that are perturbed by these variants, we generate a large transcriptome data set (nine disease-relevant cell types) and identify 23,650 cis-eQTL. We show that these are determined by â¼9720 regulatory modules, of which â¼3000 operate in multiple tissues and â¼970 on multiple genes. We identify regulatory modules that drive the disease association for 63 of the 200 risk loci, and show that these are enriched in multigenic modules. Based on these analyses, we resequence 45 of the corresponding 100 candidate genes in 6600 Crohn disease (CD) cases and 5500 controls, and show with burden tests that they include likely causative genes. Our analyses indicate that ≥10-fold larger sample sizes will be required to demonstrate the causality of individual genes using this approach.