Targeted Resequencing and Functional Testing Identifies Low-Frequency Missense Variants in the Gene Encoding GARP as Significant Contributors to Atopic Dermatitis Risk

J Invest Dermatol. 2016 Dec;136(12):2380-2386. doi: 10.1016/j.jid.2016.07.009. Epub 2016 Jul 21.

Abstract

Gene-mapping studies have consistently identified a susceptibility locus for atopic dermatitis and other inflammatory diseases on chromosome band 11q13.5, with the strongest association observed for a common variant located in an intergenic region between the two annotated genes C11orf30 and LRRC32. Using a targeted resequencing approach we identified low-frequency and rare missense mutations within the LRRC32 gene encoding the protein GARP, a receptor on activated regulatory T cells that binds latent transforming growth factor-β. Subsequent association testing in more than 2,000 atopic dermatitis patients and 2,000 control subjects showed a significant excess of these LRRC32 variants in individuals with atopic dermatitis. Structural protein modeling and bioinformatic analysis predicted a disruption of protein transport upon these variants, and overexpression assays in CD4+CD25- T cells showed a significant reduction in surface expression of the mutated protein. Consistently, flow cytometric (FACS) analyses of different T-cell subtypes obtained from atopic dermatitis patients showed a significantly reduced surface expression of GARP and a reduced conversion of CD4+CD25- T cells into regulatory T cells, along with lower expression of latency-associated protein upon stimulation in carriers of the LRRC32 A407T variant. These results link inherited disturbances of transforming growth factor-β signaling with atopic dermatitis risk.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Case-Control Studies
  • Cells, Cultured
  • Chromosome Mapping
  • Dermatitis, Atopic / drug therapy
  • Dermatitis, Atopic / genetics*
  • Dermatitis, Atopic / pathology*
  • Disease Progression
  • Female
  • Gene Expression Regulation
  • Genetic Predisposition to Disease*
  • Genotype
  • Humans
  • Immunohistochemistry
  • Male
  • Membrane Proteins / genetics*
  • Molecular Sequence Data
  • Mutation, Missense*
  • Prognosis
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Risk Assessment
  • Sequence Deletion
  • T-Lymphocytes, Regulatory / immunology
  • T-Lymphocytes, Regulatory / pathology

Substances

  • LRRC32 protein, human
  • Membrane Proteins
  • RNA, Messenger