Development and Application of a Functional Human Esophageal Mucosa Explant Platform to Eosinophilic Esophagitis

Abstract

There is an increasing prevalence of esophageal diseases but intact human tissue platforms to study esophageal function, disease mechanisms, and the interactions between cell types in situ are lacking. To address this, we utilized full thickness human donor esophagi to create and validate the ex vivo function of mucosa and smooth muscle (n = 25). Explanted tissue was tested for contractile responses to carbachol and histamine. We then treated ex vivo human esophageal mucosa with a cytokine cocktail to closely mimic the Th2 and inflammatory milieu of eosinophilic esophagitis (EoE) and assessed alterations in smooth muscle and extracellular matrix function and stiffening. We found that full thickness human esophagus as well as the individual layers of circular and longitudinal muscularis propria developed tension in response to carbachol ex vivo and that mucosa demonstrated squamous cell differentiation. Treatment of mucosa with Th2 and fibrotic cytokines recapitulated the majority of the clinical Eosinophilic Esophagitis Diagnostic Profile (EDP) on fluidic transcriptional microarray. Transforming growth factor-beta-1 (TGFβ1) increased gene expression of fibronectin, smooth muscle actin, and phospholamban (p < 0.001). The EoE cocktail also increased stiffness and decreased mucosal compliance, akin to the functional alterations in EoE (p = 0.001). This work establishes a new, transcriptionally intact and physiologically functional human platform to model esophageal tissue responses in EoE.

Acquired Esophageal Strictures in Children: Morphometric and Immunohistochemical Analyses

“…19 , 20 Both primary esophageal fibroblasts and smooth muscle cells secrete fibronectin at baseline culture conditions and increase secretion following stimulation. 21 , 22 Fibronectin and periostin were detected immunohistochemically in visceral smooth muscle but not normal submucosa in CE, but were markedly reduced or absent in visceral smooth muscle in ES (not shown), consistent with ES smooth muscle having secreted them into the extracellular matrix in the architecturally altered areas in ES. Fibronectin was identified in the extracellular matrix of ES in addition to periostin, and esophageal fibroblasts and smooth muscle cells could have been sources.…”

“…24 TGFb also mediates the increased secretion of fibronectin by primary esophageal fibroblasts or smooth muscle cells co-cultured with eosinophils or eosinophil products. 18,21,22 Secretion of TGFb may have increased muscle cell contractility and fibronectin secretion in the ES in this study since it was detected immunohistochemically in cells consistent with mast cells.…”

“…15 , 18 That binding may further increase fibronectin secretion: in vitro, primary esophageal fibroblasts and smooth muscle cells co-cultured with eosinophils or exposed to eosinophil products increase fibronectin secretion. 21 , 22…”

“…15,18 That binding may further increase fibronectin secretion: in vitro, primary esophageal fibroblasts and smooth muscle cells co-cultured with eosinophils or exposed to eosinophil products increase fibronectin secretion. 21,22 Accumulation of Esophageal Mast Cells, Eosinophils and Pro-Inflammatory Cytokines Mast cells were significantly increased in endogenous muscle layers in ES. Experimental evidence links mast cells to smooth muscle proliferation and contractility.…”

Potential Value of Circular RNA circTBC1D4 in Gastrointestinal Stromal Tumors

“…Therefore, blocking this mechanism may affect the progress of gastrointestinal stromal tumor [ 24 ]. Kurten et al [ 25 ] found that the gene expression of fibronectin and smooth muscle actin was risen in eosinophilic esophagitis. The degree of ATP2A2 inhibition was determined by PLN oligomeric state, and phosphorylation of phosphatidylinositol could reduce the inhibition of ATP2A2.…”