Crosstalk between inflammation and autophagy in CeD organoids.

Celiac disease (CeD), a small bowel disorder characterized by mucosal inflammation, villous atrophy and crypt hyperplasia, is induced by gliadin peptides in genetically predisposed individuals. Alterations in inflammation, intracellular vesicular trafficking, mTOR signalling and autophagy have been reported in the intestinal epithelium of CeD patients. The aim of this manuscript was to investigate the interplay between autophagy and inflammation in intestinal mucosa of CeD patients. Increased levels of markers related to impaired autophagic flux, mTOR pathway activation and inflammatory signalling were observed in CeD biopsies and organoids from gluten-containing-diet-CeD (GCD-CeD) and gluten-free-diet-CeD (GFD-CeD) compared with controls (CTRs). Immunofluorescence staining confirmed an increase in p62 and miR-30a was decreased in CeD patients. Bafilomycin and rapamycin were used to respectively block and induce autophagic flux in intestinal organoids. Bafilomycin increased p62, pE4-BP1 and pNF-κB levels in CTRs, whereas rapamycin reduced p62, pE4-BP1 and pNF-κB levels in CeD. Bio-Plex analysis of CMs (culture medium) from CeD organoids revealed increased levels of 17/27 in CMs from GCD-CeD and of 5/27 in CMs of GFD-CeD. Control organoids treated with CMs from GCD-CeD showed a reduction in autophagic flux and an increase of inflammation, compared with the untreated CTRs. Overall, our findings support the presence of a crosstalk between autophagy and inflammation at the epithelial level in both intestinal biopsies and organoids from CeD patients.