Abstract
Anterior gradient 2 (AGR2) is a protein disulfide isomerase that is important for protein processing in the endoplasmic reticulum and is essential for mucin production in the digestive and respiratory tracts. Bi-allelic AGR2 variants were recently found to cause recurrent respiratory infections and failure to thrive with or without diarrhea (RIFTD; MIM # 620233), although the mechanisms behind this condition remain unclear. To date, at least 15 patients with homozygous AGR2 variants have been reported. Here, we report two affected siblings in a consanguineous family who had recurrent respiratory infections and digestive symptoms, one of whom needed lung transplantation. To identify the genetic cause of their symptoms, we performed exome sequencing and identified a novel homozygous missense variant in AGR2 (NM_006408.4, c.250A>C, p.(Ser84Arg)) in both affected siblings. Both parents had the identical variant in a heterozygous state. This variant is quite rare in the general population and is clinically compatible with RIFTD, substituting a highly conserved CXXS motif with CXXR. We performed structural modeling and functional studies to investigate the effect of this variant. Through transient overexpression, Ser84Arg AGR2 decreased protein stability, and promoted aberrant dimerization under non-reducing conditions. AGR2 functions in a monomer-dimer equilibrium. Size-exclusion chromatography showed that the Ser84Arg mutant had a larger molecular size than the wild-type protein under non-reducing, but not reducing conditions, indicating that Ser84Arg enhanced intermolecular disulfide bonds. In conclusion, we identified a novel pathogenic AGR2 variant and indicated its abnormal monomer-dimer equilibrium as a possible mechanism involved in the pathogenesis of RIFTD.