{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,10]],"date-time":"2026-05-10T15:07:34Z","timestamp":1778425654623,"version":"3.51.4"},"reference-count":39,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,1,12]],"date-time":"2023-01-12T00:00:00Z","timestamp":1673481600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"\u201cFondi di Ateneo per la Ricerca di Base\u201d, University of Salerno, Italy","award":["ORSA184343"],"award-info":[{"award-number":["ORSA184343"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Celiac disease (CD) is an inflammatory intestinal disease caused by the ingestion of gluten-containing cereals by genetically predisposed individuals. Constitutive differences between cells from CD patients and control subjects, including levels of protein phosphorylation, alterations of vesicular trafficking, and regulation of type 2 transglutaminase (TG2), have been reported. In the present work, we investigated how skin-derived fibroblasts from CD and control subjects responded to thapsigargin, an endoplasmic reticulum ER stress inducer, in an attempt to contribute to the comprehension of molecular features of the CD cellular phenotype. We analyzed Ca2+ levels by single-cell video-imaging and TG2 activity by a microplate assay. Western blots and PCR analyses were employed to monitor TG2 levels and markers of ER stress and autophagy. We found that the cytosolic and ER Ca2+ level of CD cells was lower than in control cells. Treatments with thapsigargin differently activated TG2 in control and CD cells, as well as caused slightly different responses regarding the activation of ER stress and the expression of autophagic markers. On the whole, our findings identified further molecular features of the celiac cellular phenotype and highlighted that CD cells appeared less capable of adapting to a stress condition and responding in a physiological way.<\/jats:p>","DOI":"10.3390\/ijms24021495","type":"journal-article","created":{"date-parts":[[2023,1,12]],"date-time":"2023-01-12T04:29:38Z","timestamp":1673497778000},"page":"1495","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Peculiar Ca2+ Homeostasis, ER Stress, Autophagy, and TG2 Modulation in Celiac Disease Patient-Derived Cells"],"prefix":"10.3390","volume":"24","author":[{"given":"Silvia","family":"Sposito","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biology, University of Salerno, 84084 Fisciano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5054-4098","authenticated-orcid":false,"given":"Agnese","family":"Secondo","sequence":"additional","affiliation":[{"name":"Division of Pharmacology, Department of Neuroscience, Reproductive and Odontostomatological Sciences, School of Medicine, University Federico II, 80138 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5858-860X","authenticated-orcid":false,"given":"Antonio Massimiliano","family":"Romanelli","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biology, University of Salerno, 84084 Fisciano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonio","family":"Montefusco","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biology, University of Salerno, 84084 Fisciano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2569-4606","authenticated-orcid":false,"given":"Merlin","family":"Nanayakkara","sequence":"additional","affiliation":[{"name":"Department of Translational Medical Science, University Federico II, 80138 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Salvatore","family":"Auricchio","sequence":"additional","affiliation":[{"name":"European Laboratory for the Investigation of Food-Induced Diseases (ELFID), University Federico II, 80138 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6190-4917","authenticated-orcid":false,"given":"Maria Vittoria","family":"Barone","sequence":"additional","affiliation":[{"name":"Department of Translational Medical Science, University Federico II, 80138 Naples, Italy"},{"name":"European Laboratory for the Investigation of Food-Induced Diseases (ELFID), University Federico II, 80138 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2825-9711","authenticated-orcid":false,"given":"Ivana","family":"Caputo","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biology, University of Salerno, 84084 Fisciano, Italy"},{"name":"European Laboratory for the Investigation of Food-Induced Diseases (ELFID), University Federico II, 80138 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2188-3829","authenticated-orcid":false,"given":"Gaetana","family":"Paolella","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biology, University of Salerno, 84084 Fisciano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1146\/annurev.immunol.18.1.53","article-title":"Molecular basis of celiac disease","volume":"18","author":"Sollid","year":"2000","journal-title":"Annu. Rev. Immunol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"674313","DOI":"10.3389\/fimmu.2021.674313","article-title":"Interplay between Gluten, HLA, Innate and Adaptive Immunity Orchestrates the Development of Coeliac Disease","volume":"12","author":"Voisine","year":"2021","journal-title":"Front. Immunol."},{"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","unstructured":"Caio, C., Volta, U., Sapone, A., Leffler, D.A., De Giorgio, R., Catassi, C., and Alessio, F. (2019). Celiac disease: A comprehensive current review. BMC Med., 17.","DOI":"10.1186\/s12916-019-1380-z"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1007\/s00251-017-0985-7","article-title":"The roles of MHC class II genes and post-translational modification in celiac disease","volume":"69","author":"Sollid","year":"2017","journal-title":"Immunogenetics"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1038\/nrm1014","article-title":"Transglutaminases: Crosslinking enzymes with pleiotropic functions","volume":"4","author":"Lorand","year":"2003","journal-title":"Nat. Rev. Mol. Cell. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"102349","DOI":"10.1016\/j.sbi.2022.102349","article-title":"Structural bases of T cell antigen receptor recognition in celiac disease","volume":"74","author":"Ciacchi","year":"2022","journal-title":"Curr. Opin. Struct. Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1136\/gut.41.6.851","article-title":"Auto-antibodies in celiac disease: Tissue transglutaminase\u2013guilt by association?","volume":"41","author":"Sollid","year":"1997","journal-title":"Gut"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Paolella, G., Nanayakkara, M., Sposito, S., Lepretti, M., Auricchio, S., Esposito, C., Barone, M.V., Martucciello, S., and Caputo, I. (2020). Constitutive differential features of type 2 transglutaminase in cells derived from celiac patients and from healthy subjects. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21041231"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"20518","DOI":"10.3390\/ijms151120518","article-title":"Gliadin peptides as triggers of the proliferative and stress\/innate immune response of the celiac small intestinal mucosa","volume":"15","author":"Barone","year":"2014","journal-title":"Int. J. Mol. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/bs.ircmb.2020.10.003","article-title":"The gliadin p31-43 peptide: Inducer of multiple proinflammatory effects","volume":"358","author":"Chirdo","year":"2021","journal-title":"Int. Rev. Cell Mol. Biol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Caputo, I., Secondo, A., Lepretti, M., Paolella, G., Auricchio, S., Barone, M.V., and Esposito, C. (2012). Gliadin peptides induce tissue transglutaminase activation and ER-stress through Ca2+ mobilization in Caco-2 cells. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0045209"},{"key":"ref_13","first-page":"228","article-title":"Type 2 transglutaminase is involved in the autophagy-dependent clearance of ubiquitinated proteins","volume":"19","author":"Farrace","year":"2012","journal-title":"Cell Death Differ."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Barone, M.V., Auricchio, R., Nanayakkara, M., Greco, L., Troncone, R., and Auricchio, S. (2022). Pivotal Role of Inflammation in Celiac Disease. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23137177"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Nanayakkara, M., Lania, G., Maglio, M., Kosova, R., Sarno, M., Gaito, A., Discepolo, V., Troncone, R., Auricchio, S., and Auricchio, R. (2013). Enterocyte proliferation and signaling are constitutively altered in celiac disease. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0076006"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1038\/s42003-019-0443-1","article-title":"Constitutive alterations in vesicular trafficking increase the sensitivity of cells from celiac disease patients to gliadin","volume":"2","author":"Lania","year":"2019","journal-title":"Commun. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Discepolo, V., Lania, G., Ten Eikelder, M.L.G., Nanayakkara, M., Sepe, L., Tufano, R., Troncone, R., Auricchio, S., Auricchio, R., and Paolella, G. (2021). Pediatric Celiac Disease Patients Show Alterations of Dendritic Cell Shape and Actin Rearrangement. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22052708"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2003526","DOI":"10.1002\/advs.202003526","article-title":"In Well-Treated Celiac Patients Low-Level Mucosal Inflammation Predicts Response to 14-day Gluten Challenge","volume":"8","author":"Stamnaes","year":"2021","journal-title":"Adv. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Porpora, M., Conte, M., Lania, G., Bellomo, C., Rapacciuolo, L., Chirdo, F.G., Auricchio, R., Troncone, R., Auricchio, S., and Barone, M.V. (2022). Inflammation Is Present, Persistent and More Sensitive to Proinflammatory Triggers in Celiac Disease Enterocytes. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23041973"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1007\/s00726-016-2307-z","article-title":"Celiac anti-type 2 transglutaminase antibodies induce differential effects in fibroblasts from celiac disease patients and from healthy subjects","volume":"49","author":"Paolella","year":"2017","journal-title":"Amino Acids"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Paolella, G., Sposito, S., Romanelli, A.M., and Caputo, I. (2022). Type 2 Transglutaminase in Coeliac Disease: A Key Player in Pathogenesis, Diagnosis and Therapy. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23147513"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Jaskulska, A., Janecka, A.E., and Gach-Janczak, K. (2020). Thapsigargin-From Traditional Medicine to Anticancer Drug. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22010004"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3460","DOI":"10.1016\/j.bbamcr.2013.06.028","article-title":"ER stress-induced cell death mechanisms","volume":"1833","author":"Sano","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3344","DOI":"10.3748\/wjg.v26.i24.3344","article-title":"Intestinal Ca2+ absorption revisited: A molecular and clinical approach","volume":"26","author":"Areco","year":"2020","journal-title":"World J. Gastroenterol."},{"key":"ref_25","first-page":"1480","article-title":"Hypocalciuria in overt and subclinical celiac disease","volume":"90","author":"Ciacci","year":"1995","journal-title":"Am. J. Gastroenterol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2183","DOI":"10.1016\/j.bbamcr.2017.07.008","article-title":"Regulated proteolysis as an element of ER stress and autophagy: Implications for intestinal inflammation","volume":"1864","author":"Stengel","year":"2017","journal-title":"Biochim. Biophys. Acta Mol. Cell Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"758311","DOI":"10.3389\/fmed.2021.758311","article-title":"Crosstalk between ER Stress, Autophagy and Inflammation","volume":"8","author":"Chipurupalli","year":"2021","journal-title":"Front. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1007\/978-1-59745-157-4_4","article-title":"LC3 and Autophagy","volume":"445","author":"Tanida","year":"2008","journal-title":"Methods Mol. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1007\/978-1-60761-756-3_13","article-title":"Measurement of autophagy in cells and tissues","volume":"648","author":"Tanida","year":"2010","journal-title":"Methods Mol. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/15548627.2020.1797280","article-title":"Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)","volume":"17","author":"Klionsky","year":"2021","journal-title":"Autophagy"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1576","DOI":"10.1038\/sj.cdd.4402200","article-title":"Connecting endoplasmic reticulum stress to autophagy by unfolded protein response and calcium","volume":"14","year":"2007","journal-title":"Cell Death Differ."},{"key":"ref_32","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_33","doi-asserted-by":"crossref","unstructured":"Conte, M., Nigro, F., Porpora, M., Bellomo, C., Furone, F., Budelli, A.L., Nigro, R., Barone, M.V., and Nanayakkara, M. (2022). Gliadin Peptide P31-43 Induces mTOR\/NFk\u03b2 Activation and Reduces Autophagy: The Role of Lactobacillus paracasei CBA L74 Postbiotc. Int. J. Mol Sci., 23.","DOI":"10.3390\/ijms23073655"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1515\/hsz-2018-0217","article-title":"Transglutaminase type 2 in the regulation of proteostasis","volume":"400","author":"Rossin","year":"2019","journal-title":"Biol. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1240","DOI":"10.1161\/STROKEAHA.118.024115","article-title":"ORAI1\/STIM1 interaction intervenes in stroke and in neuroprotection induced by ischemic preconditioning through store-operated calcium entry","volume":"50","author":"Secondo","year":"2019","journal-title":"Stroke"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3440","DOI":"10.1016\/S0021-9258(19)83641-4","article-title":"A new generation of Ca2+ indicators with greatly improved fluorescence properties","volume":"260","author":"Grynkiewicz","year":"1985","journal-title":"J. Biol. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"12743","DOI":"10.1074\/jbc.M110.199737","article-title":"Potent and selective inhibitors of the inositol-requiring enzyme 1 endoribonuclease","volume":"286","author":"Volkmann","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e22780","DOI":"10.1002\/jbt.22780","article-title":"The mechanism of cytotoxicity of 4-nonylphenol in a human hepatic cell line involves ER-stress, apoptosis, and mitochondrial dysfunction","volume":"35","author":"Paolella","year":"2021","journal-title":"J. Biochem. Mol. Toxicol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"830307","DOI":"10.1155\/2010\/830307","article-title":"New Methods for monitoring endoplasmic reticulum stress and the unfolded protein response","volume":"2010","author":"Samali","year":"2010","journal-title":"Int. J. Cell Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/2\/1495\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:04:04Z","timestamp":1760119444000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/2\/1495"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,12]]},"references-count":39,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["ijms24021495"],"URL":"https:\/\/doi.org\/10.3390\/ijms24021495","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,12]]}}}