title: Identification of Autophagy-Related Genes and Their Regulatory miRNAs Associated with Celiac Disease in Children.
abstract: Celiac disease (CD) is a severe genetic autoimmune disorder, affecting about one in 100 people, where the ingestion of gluten leads to damage in the small intestine. Diagnosing CD is quite complex and requires blood tests and intestinal biopsy examinations. Controversy exists regarding making the diagnosis without biopsy, due to the large spectrum of manifesting symptoms; furthermore, small-intestinal gastroscopy examinations have a relatively complex management in the pediatric population. To identify novel molecular markers useful to increase the sensitivity and specificity in the diagnosis of pediatric CD patients, the expression levels of two key autophagy executor genes (ATG7andBECN1) and their regulatory validated miRNAs (miR-17 and miR-30a, respectively) were analyzed by relative quantitative real-time-PCR on a cohort of confirmed CD patients compared to age-related controls. Among the investigated targets, the non-parametric Mann-WhitneyUtest and ROC analysis indicated the highest significant association ofBECN1with CD status in the blood, while in intestinal biopsies, all of the investigated sequences were positively associated with CD diagnosis. Nomogram-based analysis showed nearly opposite expression trends in blood compared to intestine tissue, while hierarchical clustering dendrograms enabled identifying CD and control subgroups based on specific genes and miRNA expression signatures. Next, using an established in vitro approach, through digested gliadin administration in Caco-2 cells, we also highlighted that the modulation of miR-17 endogenous levels using enriched exosomes increased the intracellular autophagosome content, thereby altering the autophagic status. Altogether, these results highlighted novel molecular markers that might be useful to increase the accuracy in CD diagnosis and in molecular-based stratification of the patients, further reinforcing the functional involvement of the regulation of the autophagy process within a digestive and autoimmune-related disorder as CD.
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy. sergio.comincini@unipv.it.
authors.forename: Sergio
authors.initials: S
authors.lastname: Comincini
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy. federico.manai01@universitadipavia.it.
authors.forename: Federico
authors.initials: F
authors.lastname: Manai
authors.affiliation: Pediatrics and Adolescentology Unit, Department of Internal Medicine and Therapeutics, University of Pavia, Fondazione IRCCS San Matteo, 27100 Pavia, Italy. c.meazza@smatteo.pv.it.
authors.forename: Cristina
authors.initials: C
authors.lastname: Meazza
authors.affiliation: Pediatrics and Adolescentology Unit, Department of Internal Medicine and Therapeutics, University of Pavia, Fondazione IRCCS San Matteo, 27100 Pavia, Italy. s.pagani@smatteo.pv.it.
authors.forename: Sara
authors.initials: S
authors.lastname: Pagani
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy. carolina.martinelli01@universitadipavia.it.
authors.forename: Carolina
authors.initials: C
authors.lastname: Martinelli
authors.affiliation: Department of Clinical-Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Fondazione IRCCS San Matteo, 27100 Pavia, Italy. noemi.pasqua@hotmail.it.
authors.forename: Noemi
authors.initials: N
authors.lastname: Pasqua
authors.affiliation: Department of Clinical-Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Fondazione IRCCS San Matteo, 27100 Pavia, Italy. g.pelizzo@smatteo.pv.it.
authors.forename: Gloria
authors.initials: G
authors.lastname: Pelizzo
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy. marco.biggiogera@unipv.it.
authors.forename: Marco
authors.initials: M
authors.lastname: Biggiogera
authors.affiliation: Pediatrics and Adolescentology Unit, Department of Internal Medicine and Therapeutics, University of Pavia, Fondazione IRCCS San Matteo, 27100 Pavia, Italy. mauro.bozzola@unipv.it.
authors.forename: Mauro
authors.initials: M
authors.lastname: Bozzola
mesh_terms: D001343:Autophagy / Q000235:genetics*; D015415:Biomarkers; D001706:Biopsy; D016022:Case-Control Studies; D002446:Celiac Disease / Q000235:genetics* / Q000276:immunology / Q000473:pathology; D002460:Cell Line; D002648:Child; D019295:Computational Biology / Q000379:methods; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism / Q000473:pathology; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D008954:Models, Biological; D034622:RNA Interference; D012372:ROC Curve
chemical_list: D015415:Biomarkers; D035683:MicroRNAs
keywords: autophagy; celiac disease molecular markers; microRNA