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

Sana Syed,Paul Kelly,Shan Guleria,Christopher Moskaluk,Davide Di Fusco,Beatrice C Amadi,S Asad Ali,Najeeha T Iqbal,Laura Di Iorio,Kamran Sadiq,Giovanni Monteleone,Vincenzo Dinallo

doi:10.1371/journal.pntd.0006224

Abstract

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.

Highlights

Environmental enteropathy (EE) is an acquired inflammatory disease of the small intestine, characterized histologically by blunted villi, elongated crypts, and increased lymphocytic infiltration of the lamina propria (LP) [1,2,3]
Environmental enteropathy (EE) is a disease found in children living in low- and middleincome countries, involving gut inflammation similar to that seen in Crohn’s disease or celiac disease
The inflammation leads to poor absorption of nutrients that these children would normally get in their diets, and this malabsorption in turn leads to poor growth, problems with cognitive development, and reduced efficacy of orally-administered vaccines

Summary

Introduction

Environmental enteropathy (EE) is an acquired inflammatory disease of the small intestine, characterized histologically by blunted villi, elongated crypts, and increased lymphocytic infiltration of the lamina propria (LP) [1,2,3]. These structural and inflammatory changes characteristic of EE are tightly linked to the malabsorption characteristic of EE. In Zambia, diarrhea duration was defined as maternal recall of number of days the child had diarrhea at the time of presentation or enrollment into the study (mean duration of 33 days).

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