High SMAD7 and p-SMAD2,3 expression is associated with environmental enteropathy in children

PLoS Negl Trop Dis. 2018 Feb 7;12(2):e0006224. doi: 10.1371/journal.pntd.0006224. eCollection 2018 Feb.


Enteropathies such as Crohn's disease are associated with enteric inflammation characterized by impaired TGF-β signaling, decreased expression of phosphorylated (p)-SMAD2,3 and increased expression of SMAD7 (an inhibitor of SMAD3 phosphorylation). Environmental enteropathy (EE) is an acquired inflammatory disease of the small intestine (SI), which is associated with linear growth disruption, cognitive deficits, and reduced oral vaccine responsiveness in children <5 y in resource-poor countries. We aimed to characterize EE inflammatory pathways by determining SMAD7 and p-SMAD2,3 levels (using Western blotting) in EE duodenal biopsies (N = 19 children, 7 from Pakistan, 12 from Zambia) and comparing these with healthy controls (Ctl) and celiac disease (CD) patients from Italy. Densitometric analysis of immunoblots showed that EE SI biopsies expressed higher levels of both SMAD7 (mean±SD in arbitrary units [a.u.], Ctl = 0.47±0.20 a.u., EE = 1.13±0.25 a.u., p-value = 0.03) and p-SMAD2,3 (mean±SD, Ctl = 0.38±0.14 a.u., EE = 0.60±0.10 a.u., p-value = 0.03). Immunohistochemistry showed that, in EE, SMAD7 is expressed in both the epithelium and in mononuclear cells of the lamina propria (LP). In contrast, p-SMAD3 in EE is expressed much more prominently in epithelial cells than in the LP. The high SMAD7 immunoreactivity and lack of p-SMAD3 expression in the LP suggests defective TGF-β signaling in the LP in EE similar to a previously reported SMAD7-mediated inflammatory pathway in refractory CD and Crohn's disease. However, Western blot densitometry showed elevated p-SMAD2,3 levels in EE, possibly suggesting a different inflammatory pathway than Crohn's disease but more likely reflecting cumulative protein expression from across all compartments of the mucosa as opposed to the LP alone. Further studies are needed to substantiate these preliminary results and to illustrate the relationship between SMAD proteins, TGF-β signaling, and inflammatory cytokine production, all of which may be potential therapeutic targets.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Biopsy
  • Child
  • Child, Preschool
  • Duodenum / pathology
  • Endoscopy
  • Epithelial Cells / metabolism
  • Epithelium / metabolism
  • Female
  • Gene Expression Regulation
  • Humans
  • Immunohistochemistry
  • Infant
  • Italy
  • Male
  • Mucous Membrane / metabolism
  • Pakistan
  • Phosphorylation
  • Signal Transduction
  • Smad2 Protein / metabolism*
  • Smad3 Protein / metabolism*
  • Smad7 Protein / metabolism*
  • Transforming Growth Factor beta / metabolism
  • Vaccines
  • Zambia


  • SMAD2 protein, human
  • SMAD3 protein, human
  • SMAD7 protein, human
  • Smad2 Protein
  • Smad3 Protein
  • Smad7 Protein
  • Transforming Growth Factor beta
  • Vaccines

Grants and funding

This work was supported by grants from the Bill & Melinda Gates Foundation (SAA: OPP1066200, PK: OPP1066118, GM: OPP1131242). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.