Human AGR2 Deficiency Causes Mucus Barrier Dysfunction and Infantile Inflammatory Bowel Disease

Cell Mol Gastroenterol Hepatol. 2021;12(5):1809-1830. doi: 10.1016/j.jcmgh.2021.07.001. Epub 2021 Jul 6.

Abstract

Background & aims: The gastrointestinal epithelium plays a crucial role in maintaining homeostasis with the gut microbiome. Mucins are essential for intestinal barrier function and serve as a scaffold for antimicrobial factors. Mucin 2 (MUC2) is the major intestinal gel-forming mucin produced predominantly by goblet cells. Goblet cells express anterior gradient 2 (AGR2), a protein disulfide isomerase that is crucial for proper processing of gel-forming mucins. Here, we investigated 2 siblings who presented with severe infantile-onset inflammatory bowel disease.

Methods: We performed whole-genome sequencing to identify candidate variants. We quantified goblet cell numbers using H&E histology and investigated the expression of gel-forming mucins, stress markers, and goblet cell markers using immunohistochemistry. AGR2-MUC2 binding was evaluated using co-immunoprecipitation. Endoplasmic reticulum (ER) stress regulatory function of mutant AGR2 was examined by expression studies in Human Embryonic Kidney 293T (HEK293T) using tunicamycin to induce ER stress.

Results: Both affected siblings were homozygous for a missense variant in AGR2. Patient biopsy specimens showed reduced goblet cells; depletion of MUC2, MUC5AC, and MUC6; up-regulation of AGR2; and increased ER stress. The mutant AGR2 showed reduced capacity to bind MUC2 and alleviate tunicamycin-induced ER stress.

Conclusions: Phenotype-genotype segregation, functional experiments, and the striking similarity of the human phenotype to AGR2-/- mouse models suggest that the AGR2 missense variant is pathogenic. The Mendelian deficiency of AGR2, termed "Enteropathy caused by AGR2 deficiency, Goblet cell Loss, and ER Stress" (EAGLES), results in a mucus barrier defect, the inability to mitigate ER stress, and causes infantile-onset inflammatory bowel disease.

Keywords: AGR2; ER Stress; Goblet Cells; Intestinal Metaplasia; MUC2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Biomarkers
  • Disease Models, Animal
  • Disease Susceptibility*
  • Endoplasmic Reticulum Stress
  • Gastric Mucosa / metabolism
  • Gastric Mucosa / pathology
  • Genetic Predisposition to Disease
  • Goblet Cells / metabolism
  • Goblet Cells / pathology
  • Humans
  • Inflammatory Bowel Diseases / diagnosis
  • Inflammatory Bowel Diseases / etiology*
  • Inflammatory Bowel Diseases / metabolism*
  • Intestinal Mucosa / metabolism*
  • Intestinal Mucosa / pathology
  • Male
  • Mice
  • Mice, Knockout
  • Mucins / genetics
  • Mucins / metabolism
  • Mucoproteins / chemistry
  • Mucoproteins / deficiency*
  • Mucoproteins / metabolism
  • Mucus / metabolism*
  • Oncogene Proteins / chemistry
  • Oncogene Proteins / deficiency*
  • Oncogene Proteins / metabolism
  • Phenotype
  • Sequence Analysis, DNA
  • Siblings
  • Structure-Activity Relationship
  • Whole Genome Sequencing

Substances

  • AGR2 protein, human
  • Biomarkers
  • Mucins
  • Mucoproteins
  • Oncogene Proteins