Serena Magni1, Gaia Buoli Comani1, Luca Elli2, Samanta Vanessi1, Elisa Ballarini3, Gabriella Nicolini3, Michela Rusconi1, Mirco Castoldi4, Raffaella Meneveri1, Martina U Muckenthaler5, Maria Teresa Bardella2, Donatella Barisani6. 1. Department of Health Sciences, School of Medicine, University of Milano-Bicocca, Monza, Italy. 2. Center for the Prevention and Diagnosis of Celiac Disease and UOC Gastroenterologia ed Endoscopia, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti, Università degli Studi di Milano, Milan, Italy. 3. Experimental Neurology Unit, Department of Surgery and Translational Medicine, School of Medicine, University of Milano-Bicocca, Monza, Italy. 4. 1] Department of Pediatric Oncology, Hematology and Immunology, University Hospital of Heidelberg, Heidelberg, Germany [2] Clinic for Gastroenterology, Hepatology and Infectious Diseases, Heinrich-Heine University Düsseldorf, Düsseldorf, Germany. 5. Department of Pediatric Oncology, Hematology and Immunology, University Hospital of Heidelberg, Heidelberg, Germany. 6. 1] Department of Health Sciences, School of Medicine, University of Milano-Bicocca, Monza, Italy [2] International Research Center for Hepato-Biliary-Pancreatic Diseases-IRCHD, University of Milano-Bicocca, Monza, Italy.
Abstract
OBJECTIVES: microRNAs (miRNAs) are short RNAs that regulate gene expression in various processes, including immune response. Altered immune response is a pivotal event in the pathogenesis of celiac disease (CD), and miRNAs could have a role in modulating both innate and adaptive response to gluten in celiac patients. METHODS: We compared miRNA profiles in duodenal biopsies of controls and CD patients by miRNA array. Differentially expressed miRNAs were validated in controls, Marsh 3A-B, and Marsh 3C patients by quantitative PCR (qPCR). Target gene expression was assessed by qPCR, western blotting, and immunohistochemistry, and the effect of gliadin was evaluated by in vitro stimulation experiments on duodenal biopsies. RESULTS: Seven miRNAs were identified as significantly downregulated in the duodenum of adult CD patients as compared with controls. qPCR validated the decreased expression of miR-192-5p, miR-31-5p, miR-338-3p, and miR-197, in particular in patients with more severe histological lesions (Marsh 3C). In silico analysis of possible miRNA targets identified several genes involved in innate and adaptive immunity. Among these, chemokine C-X-C motif ligand 2 (CXCL2) and NOD2 showed significantly increased mRNA and protein level in Marsh 3C patients and a significant inverse correlation with the regulatory miR-192-5p. In addition, forkhead box P3 (FOXP3), Run-related transcription factor 1, and interleukin-18 (targets of miR-31-5p, miR-338-3p, and miR-197, respectively) showed upregulation in CD patients. Furthermore, alterations in CXCL2 and NOD2, FOXP3, miR-192-5p, and miR-31-5p expression were triggered by gliadin exposure in CD patients. CONCLUSIONS: miRNA expression is significantly altered in duodenal mucosa of CD patients, and this alteration can increase the expression of molecules involved in immune response.
OBJECTIVES: microRNAs (miRNAs) are short RNAs that regulate gene expression in various processes, including immune response. Altered immune response is a pivotal event in the pathogenesis of celiac disease (CD), and miRNAs could have a role in modulating both innate and adaptive response to gluten in celiac patients. METHODS: We compared miRNA profiles in duodenal biopsies of controls and CDpatients by miRNA array. Differentially expressed miRNAs were validated in controls, Marsh 3A-B, and Marsh 3C patients by quantitative PCR (qPCR). Target gene expression was assessed by qPCR, western blotting, and immunohistochemistry, and the effect of gliadin was evaluated by in vitro stimulation experiments on duodenal biopsies. RESULTS: Seven miRNAs were identified as significantly downregulated in the duodenum of adult CDpatients as compared with controls. qPCR validated the decreased expression of miR-192-5p, miR-31-5p, miR-338-3p, and miR-197, in particular in patients with more severe histological lesions (Marsh 3C). In silico analysis of possible miRNA targets identified several genes involved in innate and adaptive immunity. Among these, chemokine C-X-C motif ligand 2 (CXCL2) and NOD2 showed significantly increased mRNA and protein level in Marsh 3C patients and a significant inverse correlation with the regulatory miR-192-5p. In addition, forkhead box P3 (FOXP3), Run-related transcription factor 1, and interleukin-18 (targets of miR-31-5p, miR-338-3p, and miR-197, respectively) showed upregulation in CDpatients. Furthermore, alterations in CXCL2 and NOD2, FOXP3, miR-192-5p, and miR-31-5p expression were triggered by gliadin exposure in CDpatients. CONCLUSIONS: miRNA expression is significantly altered in duodenal mucosa of CDpatients, and this alteration can increase the expression of molecules involved in immune response.
Authors: Nasim Samandari; Aashiq H Mirza; Lotte B Nielsen; Simranjeet Kaur; Philip Hougaard; Siri Fredheim; Henrik B Mortensen; Flemming Pociot Journal: Diabetologia Date: 2016-11-19 Impact factor: 10.122
Authors: Aarón D Ramírez-Sánchez; Ineke L Tan; B C Gonera-de Jong; Marijn C Visschedijk; Iris Jonkers; Sebo Withoff Journal: Int J Mol Sci Date: 2020-11-12 Impact factor: 5.923