title: Long-Range Modulation of PAG1 Expression by 8q21 Allergy Risk Variants.
abstract: The gene(s) whose expression is regulated by allergy risk variants is unknown for many loci identified through genome-wide association studies. Addressing this knowledge gap might point to new therapeutic targets for allergic disease. The aim of this study was to identify the target gene(s) and the functional variant(s) underlying the association between rs7009110 on chromosome 8q21 and allergies. Eight genes are located within 1 Mb of rs7009110. Multivariate association analysis of publicly available exon expression levels from lymphoblastoid cell lines (LCLs) identified a significant association between rs7009110 and the expression of a single gene, PAG1 (p = 0.0017), 732 kb away. Analysis of histone modifications and DNase I hypersensitive sites in LCLs identified four putative regulatory elements (PREs) in the region. Chromosome conformation capture confirmed that two PREs interacted with the PAG1 promoter, one in allele-specific fashion. To determine whether these PREs were functional, LCLs were transfected with PAG1 promoter-driven luciferase reporter constructs. PRE3 acted as a transcriptional enhancer for PAG1 exclusively when it carried the rs2370615:C allergy predisposing allele, a variant in complete linkage disequilibrium with rs7009110. As such, rs2370615, which overlaps RelA transcription factor (TF) binding in LCLs and was found to disrupt Foxo3a binding to PRE3, represents the putative functional variant in this locus. Our studies suggest that the risk-associated allele of rs2370615 predisposes to allergic disease by increasing PAG1 expression, which might promote B cell activation and have a pro-inflammatory effect. Inhibition of PAG1 expression or function might have therapeutic potential for allergic diseases.
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, QLD 4029, Australia.
authors.forename: Cristina T
authors.initials: CT
authors.lastname: Vicente
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, QLD 4029, Australia.
authors.forename: Stacey L
authors.initials: SL
authors.lastname: Edwards
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, QLD 4029, Australia.
authors.forename: Kristine M
authors.initials: KM
authors.lastname: Hillman
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, QLD 4029, Australia.
authors.forename: Susanne
authors.initials: S
authors.lastname: Kaufmann
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, QLD 4029, Australia.
authors.forename: Hayley
authors.initials: H
authors.lastname: Mitchell
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, QLD 4029, Australia.
authors.forename: Lisa
authors.initials: L
authors.lastname: Bain
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, QLD 4029, Australia.
authors.forename: Dylan M
authors.initials: DM
authors.lastname: Glubb
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, QLD 4029, Australia.
authors.forename: Jason S
authors.initials: JS
authors.lastname: Lee
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, QLD 4029, Australia. Electronic address: juliet.french@qimrberghofer.edu.au.
authors.forename: Juliet D
authors.initials: JD
authors.lastname: French
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, QLD 4029, Australia. Electronic address: manuel.ferreira@qimrberghofer.edu.au.
authors.forename: Manuel A R
authors.initials: MA
authors.lastname: Ferreira
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics*; D001402:B-Lymphocytes / Q000276:immunology; D002898:Chromosomes, Human, Pair 8 / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics*; D056726:Genetic Association Studies; D006801:Humans; D006967:Hypersensitivity / Q000235:genetics* / Q000276:immunology; D015810:Linkage Disequilibrium; D008156:Luciferases; D008213:Lymphocyte Activation / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics*; D015999:Multivariate Analysis; D009690:Nucleic Acid Conformation; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D008565:Membrane Proteins; C498710:PAG1 protein, human; D008156:Luciferases