Eosinophilic esophagitis (EoE) is a chronic inflammatory disease of the esophagus, believed to be triggered by an allergen-specific immune response, resulting in progressive barrier dysfunction and impaired wound healing. EoE is characterised by the influx of inflammatory cells such as eosinophils and leukocytes to the esophagus driving a type-2 inflammatory response in EoE, sequestering available oxygen in the esophageal epithelium and creating a pathologically hypoxic microenvironment. Previously published literature has demonstrated a significant attenuation of HIF-1α signalling in EoE patients and additionally in an in vivo L2-IL5OXA EoE mouse model, contributing to esophageal epithelial barrier dysfunction mediated via decreased claudin-1 expression within EoE. Furthermore, HIF-1α, via transcriptional upregulation of target genes is critical for mediating homeostatic epithelial functions such as cell migration, proliferation and barrier restoration, essential processes for effective wound healing responses, which have been demonstrated to be dysregulated in EoE. Building on previously published data, CD73 a direct downstream transcriptional target of HIF-1α was shown to be significantly diminished in EoE patients. CD73/ecto-5’- nucleotidase plays a critical role in the production of extracellular adenosine, a signalling molecule important for playing an adaptive and protective role during hypoxia via interaction with extracellular adenosine receptors. These studies aimed to investigate for the first time in the esophagus, the functional consequence diminished CD73 expression has on mediating esophageal epithelial dysfunction within EoE. With reference to the growing body of literature that extracellular adenosine receptor ADORA2B is critical in protecting epithelial barrier integrity and wound healing responses during hypoxia, it became an attractive novel pathway to further investigate in the esophageal epithelium. We hypothesized that diminished CD73 expression in EoE, results in altered downstream ADROA2B signalling, mediating esophageal epithelial dysfunction within EoE. In line with this, we combined pharmacologic studies and in vivo mouse model studies using a L2-IL5OXA EoE mouse model to address our hypothesis. Our studies demonstrated that pharmacological inhibition of CD73 using adenosine 5’-(ɑ-β-methylene) diphosphate (APCP) resulted in defective wound healing responses in in vitro scratch assays and dysfunctional epithelial barrier in 3-dimensional air liquid interface cultures measured by transepithelial electrical resistance (TEER). In concordance with published literature, we demonstrated for the first time in the esophagus that activation of downstream ADORA2B signalling via administration of ADORA2B agonist BAY606583 improved wound healing responses and barrier functions via restoration of fibronectin (FN1) and occludin (OCLN) expression, which have previously been shown to be associated with downstream ADORA2B signalling. Similarly, in vivo studies in L2-IL5OXA EoE mouse models recapitulated our in vitro findings of improved epithelial barrier integrity characterised by increased expression of occludin following treatment with ADORA2B specific agonist BAY606583. Overall, our studies demonstrate a significant contribution of diminished CD73 expression and altered ADORA2B signalling in mediating epithelial dysfunction within EoE via defective wound healing and barrier dysfunction and establishes an exciting possibility for ADORA2B agonist, such as BAY606583 as a potential novel therapeutic approach for EoE.