Metabolic dysfunction mediated by HIF-1α contributes to epithelial differentiation defects in eosinophilic esophagitis - 18/09/24

Doi : 10.1016/j.jaci.2024.07.030

Sinéad Ryan, PhD ^a,^b, Louise Crowe, BSc ^a,^b, Sofía N. Almeida Cruz, MSc ^a,^b, Matthew D. Galbraith, PhD ^c,^d, Carol O’Brien, PhD ^a,^b, Juliet A. Hammer, BA ^e, Ronan Bergin, PhD ^b, Shauna K. Kellett, BSc ^a,^b, Gary E. Markey, PhD ^a,^b, Taylor M. Benson, BA ^a,^b, Olga Fagan, BSc, BM, BS ^f, Joaquin M. Espinosa, PhD ^c, Niall Conlon, MB, BCh, BAO, PGDip (Allergy), PhD, MRCP (UK), FRCPath ^g, Claire L. Donohoe, MB, BCh, BAO, BA, MMEd, PhD, FRCSI ^h, Susan McKiernan, MB, BCh, BAO ^f, Andrew E. Hogan, PhD ^b,ⁱ, Eóin N. McNamee, PhD ^b,^j, Glenn T. Furuta, MD ^e, Calies Menard-Katcher, MD ^e, Joanne C. Masterson, PhD ^a,^b,^e,^⁎
^a Allergy, Inflammation, and Remodeling Research Laboratory, Department of Biology, National University of Ireland, Maynooth, Ireland
^b Kathleen Lonsdale Institute for Human Health Research, Maynooth University, Maynooth, Ireland
^c Linda Crinc Institute for Down Syndrome, University of Colorado Anschutz Medical Campus, Aurora, Colo
^d Department of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, Colo
^e Gastrointestinal Eosinophilic Diseases Program, Digestive Health Institute, Children’s Hospital Colorado, Department of Pediatrics, University of Colorado School of Medicine, Aurora, Colo
^f Department of Gastroenterology, St James’s Hospital, Dublin, Ireland
^g Department of Allergy and Immunology, St James’s Hospital, Dublin, Ireland
^h National Centre for Oesophageal and Gastric Cancer, Trinity St James’s Cancer Institute, St James’s Hospital, Trinity College, Dublin, Ireland
ⁱ Department of Biology, Obesity Immunology Research Group, Maynooth University, Maynooth, Ireland
^j Department of Biology, Mucosal Immunology Research Laboratory, National University of Ireland, Maynooth, Ireland

^∗Corresponding author: Joanne C. Masterson, PhD, Allergy, Inflammation and Remodeling Research Laboratory, Kathleen Lonsdale Institute for Human Health Research, Department of Biology, National University of Ireland, Maynooth, Co Kildare, Ireland.Inflammation and Remodeling Research LaboratoryKathleen Lonsdale Institute for Human Health ResearchDepartment of BiologyNational University of IrelandMaynoothCo KildareIreland

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Graphical abstract

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Abstract

Background

Investigating the contributory role that epithelial cell metabolism plays in allergic inflammation is a key factor to understanding what influences dysfunction and the pathogenesis of the allergic disease eosinophilic esophagitis (EoE). We previously highlighted that the absence of hypoxia signaling through hypoxia-inducible factor (HIF)-1α in EoE contributes to esophageal epithelial dysfunction. However, metabolic regulation by HIF-1α has not been explored in esophageal allergy.

Objectives

We sought to define the role of HIF-1α–mediated metabolic dysfunction in esophageal epithelial differentiation processes and barrier function in EoE.

Methods

In RNA sequencing of EoE patient biopsy samples, we observed the expression pattern of key genes involved in mitochondrial metabolism/oxidative phosphorylation (OXPHOS) and glycolysis. Seahorse bioenergetics analysis was performed on EPC2-hTERT cells to decipher the metabolic processes involved in epithelial differentiation processes. In addition, air–liquid interface cultures were used to delineate metabolic dependency mechanisms required for epithelial differentiation.

Results

Transcriptomic analysis identified an increase in genes associated with OXPHOS in patients with EoE. Epithelial origin of this signature was confirmed by complex V immunofluorescence of patient biopsy samples. Bioenergetic analysis in vitro revealed that differentiated epithelium was less reliant on OXPHOS compared with undifferentiated epithelium. Increased OXPHOS potential and reduced glycolytic capacity was mirrored in HIF1A-knockdown EPC2-hTERT cells that exhibited a significant absence of terminal markers of epithelial differentiation, including involucrin. Pharmacologic glucose transport inhibition phenocopied this, while rescue of the HIF-1α–deficient phenotype using the pan-prolyl hydroxylase inhibitor dimethyloxalylglycine resulted in restored expression of epithelial differentiation markers.

Conclusions

An OXPHOS-dominated metabolic pattern in EoE patients, brought about largely by the absence of HIF-1α–mediated glycolysis, is linked with the deficit in esophageal epithelial differentiation.

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Key words : Eosinophilic esophagitis, HIF-1α, glycolysis, differentiation

Abbreviations used : ALI, ATP, 2-DG, DMOG, ECAR, EoE, FCCP, FDR, FLG, GEO, GLUT1, GO, GSEA, HIF-1α, IVL, KD, KEGG, KRT, KSFM, NDUF, OCR, OXPHOS, PDK1, PHD, RNA-Seq, SCENITH, shCtrl, SPRR