{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T22:23:39Z","timestamp":1789424619594,"version":"build-2803163510"},"reference-count":48,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2019,4,12]],"date-time":"2019-04-12T00:00:00Z","timestamp":1555027200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Celiac disease (CD) is a chronic systemic autoimmune disorder that is triggered by the ingestion of gliadin peptides, the alcohol-soluble fraction of wheat gluten. These peptides, which play a key role in the immune response that underlies CD, spontaneously form aggregates and exert a direct toxic action on cells due to the increase in the reactive oxygen species (ROS) levels. Furthermore, peptic-tryptic digested gliadin peptides (PT-gliadin) lead to an impairment in the autophagy pathway in an in vitro model based on Caco-2 cells. Considering these premises, in this study we have analyzed different mTOR-independent inducers, reporting that the disaccharide trehalose, a mTOR-independent autophagy activator, rescued the autophagy flux in Caco-2 cells treated with digested gliadin, as well as improved cell viability. Moreover, trehalose administration to Caco-2 cells in presence of digested gliadin reduced the intracellular levels of these toxic peptides. Altogether, these results showed the beneficial effects of trehalose in a CD in vitro model as well as underlining autophagy as a molecular pathway whose modulation might be promising in counteracting PT-gliadin cytotoxicity.<\/jats:p>","DOI":"10.3390\/cells8040348","type":"journal-article","created":{"date-parts":[[2019,4,12]],"date-time":"2019-04-12T12:55:04Z","timestamp":1555073704000},"page":"348","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Trehalose Modulates Autophagy Process to Counteract Gliadin Cytotoxicity in an In Vitro Celiac Disease Model"],"prefix":"10.3390","volume":"8","author":[{"given":"Federico","family":"Manai","sequence":"first","affiliation":[{"name":"Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6037-853X","authenticated-orcid":false,"given":"Alberto","family":"Azzalin","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martina","family":"Morandi","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Veronica","family":"Riccardi","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa","family":"Zanoletti","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Dei Giudici","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabio","family":"Gabriele","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolina","family":"Martinelli","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4882-3714","authenticated-orcid":false,"given":"Mauro","family":"Bozzola","sequence":"additional","affiliation":[{"name":"Pediatrics and Adolescentology Units, Department of Internal Medicine and Therapeutics, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sergio","family":"Comincini","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,4,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1480","DOI":"10.1016\/S0140-6736(09)60254-3","article-title":"Coeliac disease","volume":"373","author":"Corazza","year":"2009","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"862","DOI":"10.1136\/gut.15.11.862","article-title":"The toxicity of certain cereal proteins in coeliac disease","volume":"15","author":"Cornell","year":"1974","journal-title":"Gut"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.fm.2006.07.004","article-title":"Chemistry of gluten proteins","volume":"24","author":"Wieser","year":"2007","journal-title":"Food Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1111\/j.1651-2227.1996.tb14239.x","article-title":"Relation between gliadin structure and coeliac toxicity","volume":"412","author":"Wieser","year":"1996","journal-title":"Acta Paediatr. Suppl."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1007\/s007260170010","article-title":"Structure-activity relationships in coeliac-toxic gliadin peptides","volume":"21","author":"Cornell","year":"2001","journal-title":"Amino Acids"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Barone, M.V., Nanayakkara, M., Paolella, G., Maglio, M., Vitale, V., Troiano, R., Ribecco, M.T., Lania, G., Zanzi, D., and Santagata, S. (2010). Gliadin peptide P31-43 localises to endocytic vesicles and interferes with their maturation. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0012246"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"8648","DOI":"10.1039\/C5SM01619C","article-title":"Self-assembly of 33-mer gliadin peptide oligomers","volume":"11","author":"Herrera","year":"2015","journal-title":"Soft Matter"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1016\/j.colsurfb.2018.02.053","article-title":"Insights into gliadin supramolecular organization at digestive pH 3.0","volume":"165","author":"Herrera","year":"2018","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1417","DOI":"10.1016\/j.nano.2018.04.014","article-title":"Large supramolecular structures of 33-mer gliadin peptide activate toll-like receptors in macrophages","volume":"14","author":"Herrera","year":"2018","journal-title":"Nanomedicine"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1159\/000342588","article-title":"Inflammatory bowel disease: Dysfunction of autophagy?","volume":"30","author":"Scharl","year":"2012","journal-title":"Dig. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1392","DOI":"10.1002\/ibd.21486","article-title":"Genetic variation in the autophagy gene ULK1 and risk of Crohn\u2019s disease","volume":"17","author":"Henckaerts","year":"2011","journal-title":"Inflamm. Bowel Dis."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Manai, F., Azzalin, A., Gabriele, F., Martinelli, C., Morandi, M., Biggiogera, M., Bozzola, M., and Comincini, S. (2018). The in vitro effects of enzymatic digested gliadin on the functionality of the autophagy process. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19020635"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Comincini, S., Manai, F., Meazza, C., Pagani, S., Martinelli, C., Pasqua, N., Pelizzo, G., Biggiogera, M., and Bozzola, M. (2017). Identification of autophagy-related genes and their regulatory miRNAs associated with celiac disease in children. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18020391"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1016\/j.molcel.2010.09.023","article-title":"Autophagy and the integrated stress response","volume":"40","author":"Kroemer","year":"2010","journal-title":"Mol. Cell"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"736905","DOI":"10.1155\/2012\/736905","article-title":"Aggrephagy: Selective disposal of protein aggregates by macroautophagy","volume":"2012","author":"Lamark","year":"2012","journal-title":"Int. J. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"7234","DOI":"10.1074\/jbc.M114.597492","article-title":"Autophagy enhances intestinal epithelial tight junction barrier function by targeting claudin-2 protein degradation","volume":"290","author":"Nighot","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1002\/humu.22529","article-title":"TBC1D7 mutations are associated with intellectual disability, macrocrania, patellar dislocation, and celiac disease","volume":"35","author":"Alfaiz","year":"2014","journal-title":"Hum. Mutat."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1007\/s11894-017-0597-2","article-title":"Going gluten free: The history and nutritional mplications of today\u2019s most popular diet","volume":"19","author":"Newberry","year":"2017","journal-title":"Curr. Gastroenterol. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"28423","DOI":"10.1038\/srep28423","article-title":"Trehalose, sucrose and raffinose are novel activators of autophagy in human keratinocytes through an mTOR-independent pathway","volume":"6","author":"Chen","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.redox.2017.09.007","article-title":"Trehalose protects against oxidative stress by regulating the Keap1-Nrf2 and autophagy pathways","volume":"15","author":"Mizunoe","year":"2018","journal-title":"Redox. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5641","DOI":"10.1074\/jbc.M609532200","article-title":"Trehalose, a novel mTOR-independent autophagy enhancer, accelerates the clearance of mutant huntingtin and alpha-synuclein","volume":"282","author":"Sarkar","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1080\/00365520500235334","article-title":"Gliadin, zonulin and gut permeability: Effects on celiac and non-celiac intestinal mucosa and intestinal cell lines","volume":"41","author":"Drago","year":"2006","journal-title":"Scand. J. Gastroenterol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/15548627.2015.1100356","article-title":"Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)","volume":"12","author":"Klionsky","year":"2016","journal-title":"Autophagy"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"736","DOI":"10.1016\/S0016-5085(89)80072-1","article-title":"Characterization of the human colon carcinoma cell line (Caco-2) as a model system for intestinal epithelial permeability","volume":"96","author":"Hidalgo","year":"1989","journal-title":"Gastroenterology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s10565-005-0085-6","article-title":"The Caco-2 cell line as a model of the intestinal barrier: Influence of cell and culture-related factors on Caco-2 cell functional characteristics","volume":"21","author":"Sambuy","year":"2005","journal-title":"Cell Biol. Toxicol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"8832","DOI":"10.1073\/pnas.1405723111","article-title":"Contact inhibition and high cell density deactivate the mammalian target of rapamycin pathway, thus suppressing the senescence program","volume":"111","author":"Leontieva","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_27","first-page":"55","article-title":"Determination of autophagy in the Caco-2 spontaneously differentiating gmodelof intestinal epithelial cells","volume":"1854","author":"Banerjee","year":"2019","journal-title":"Methods Mol. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1369\/0022155412453052","article-title":"Imaging flow cytometry: Coping with heterogeneity in biological systems","volume":"60","author":"Barteneva","year":"2012","journal-title":"J. Histochem. Cytochem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1038\/cdd.2014.150","article-title":"Oxidative stress and autophagy: The clash between damage and metabolic needs","volume":"22","author":"Filomeni","year":"2015","journal-title":"Cell Death Differ."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1016\/j.cellsig.2013.11.028","article-title":"The role of cell signalling in the crosstalk between autophagy and apoptosis","volume":"26","author":"Booth","year":"2014","journal-title":"Cell Signal."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1016\/j.neuron.2017.01.022","article-title":"Autophagy and neurodegeneration: Pathogenic mechanisms and therapeutic opportunities","volume":"93","author":"Menzies","year":"2017","journal-title":"Neuron"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1038\/s41419-017-0252-8","article-title":"Pharmacological activation of autophagy favors the clearing of intracellular aggregates of misfolded prion protein peptide to prevent neuronal death","volume":"9","author":"Thellung","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1002\/cbin.10874","article-title":"The toxic alpha-gliadin peptide 31-43 enters cells without a surface membrane receptor","volume":"42","author":"Paolella","year":"2018","journal-title":"Cell Biol. Int."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1053\/j.gastro.2015.06.046","article-title":"Role of autophagy in the maintenance of intestinal homeostasis","volume":"149","author":"Baxt","year":"2015","journal-title":"Gastroenterology"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2501","DOI":"10.1111\/bph.13860","article-title":"Stimulation of autophagy prevents intestinal mucosal inflammation and ameliorates murine colitis","volume":"174","author":"Salvador","year":"2017","journal-title":"Br. J. Pharmacol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1007\/s12154-008-0003-5","article-title":"Rapamycin and mTOR kinase inhibitors","volume":"1","author":"Ballou","year":"2008","journal-title":"J. Chem. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1038\/nrm3293","article-title":"Sirtuins as regulators of metabolism and healthspan","volume":"13","author":"Houtkooper","year":"2012","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1186\/s13046-018-0731-5","article-title":"Metformin induces autophagy and G0\/G1 phase cell cycle arrest in myeloma by targeting the AMPK\/mTORC1 and mTORC2 pathways","volume":"37","author":"Wang","year":"2018","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1038\/nchembio883","article-title":"Small molecules enhance autophagy and reduce toxicity in Huntington\u2019s disease models","volume":"3","author":"Sarkar","year":"2007","journal-title":"Nat. Chem. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Casarejos, M.J., Perucho, J., L\u00f3pez-Send\u00f3n, J.L., Garc\u00eda de Y\u00e9benes, J., Bettencourt, C., G\u00f3mez, A., Ruiz, C., Heutink, P., Rizzu, P., and Mena, M.A. (2014). Trehalose improves human fibroblast deficits in a new CHIP-mutation related ataxia. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0106931"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Kang, Y.L., Saleem, M.A., Chan, K.W., Yung, B.Y., and Law, H.K. (2014). Trehalose, an mTOR independent autophagy inducer, alleviates human podocyte injury after puromycin aminonucleoside treatment. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0113520"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1002\/jcb.25012","article-title":"Sucrose induces vesicles accumulation and autophagy","volume":"116","author":"Higuchi","year":"2015","journal-title":"J. Cell. Biochem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2657","DOI":"10.1590\/0001-3765201720160659","article-title":"High consumption of sucrose induces DNA damage in male Wistar rats","volume":"89","author":"Franke","year":"2017","journal-title":"Anais da Academia Brasileira de Ci\u00eancias"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"17R","DOI":"10.1093\/glycob\/cwg047","article-title":"New insights on trehalose: A multifunctional molecule","volume":"13","author":"Elbein","year":"2003","journal-title":"Glycobiology"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1038\/nm985","article-title":"Trehalose alleviates polyglutamine-mediated pathology in a mouse model of Huntington disease","volume":"10","author":"Tanaka","year":"2004","journal-title":"Nat. Med."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1080\/15548627.2015.1071759","article-title":"Modulation of inflammation by autophagy: Consequences for human disease","volume":"12","author":"Plantinga","year":"2016","journal-title":"Autophagy"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.intimp.2013.11.001","article-title":"Autophagy in inflammation, infection, neurodegeneration and cancer","volume":"18","author":"Arroyo","year":"2014","journal-title":"Int. Immunopharmacol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1793","DOI":"10.1007\/s00726-011-0899-x","article-title":"TG2 transamidating activity acts as a reostat controlling the interplay between apoptosis and autophagy","volume":"42","author":"Rossin","year":"2012","journal-title":"Amino Acids"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/8\/4\/348\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:44:57Z","timestamp":1760186697000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/8\/4\/348"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,4,12]]},"references-count":48,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2019,4]]}},"alternative-id":["cells8040348"],"URL":"https:\/\/doi.org\/10.3390\/cells8040348","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,4,12]]}}}