Abstract
We have recently found that celiac disease patient serum-derived autoantibodies targeted against transglutaminase 2 interfere with several steps of angiogenesis, including endothelial sprouting and migration, though the mechanism involved remained to be fully characterized. This study now investigated the processes underlying the antiangiogenic effects exerted by celiac disease patient antibodies on endothelial cells, with particular regard to the adhesion, migration, and polarization signaling pathway. We observed that celiac IgA reduced endothelial cell numbers by affecting adhesion without increasing apoptosis. Endothelial cells in the presence of celiac IgA showed weak attachment, a high susceptibility to detach from fibronectin, and a disorganized extracellular matrix due to a reduction of protein cross-links. Furthermore, celiac patient IgA led to secretion of active transglutaminase 2 from endothelial cells into the culture supernatants. Additionally, cell surface transglutaminase 2 mediated integrin clustering in the presence of celiac IgA was coupled to augmented expression of β1-integrin. We also observed that celiac patient IgA-treated endothelial cells had migratory defects and a less polarized phenotype when compared to control groups, and this was associated with the RhoA signaling pathway. These biological effects mediated by celiac IgA on endothelial cells were partially influenced but not completely abolished by R281, an irreversible extracellular transglutaminase 2 enzymatic activity inhibitor. Taken together, our results imply that celiac patient IgA antibodies disturb the extracellular protein cross-linking function of transglutaminase 2, thus altering cell-extracellular matrix interactions and thereby affecting endothelial cell adhesion, polarization, and motility.
Similar content being viewed by others
References
Martin CA, Perrone EE, Longshore SW, Toste P, Bitter K, Nair R, Guo J, Erwin CR, Warner BW (2009) Intestinal resection induces angiogenesis within adapting intestinal villi. J Pediatr Surg 44:1077–1082
Myrsky E, Caja S, Simon-Vecsei Z, Korponay-Szabo IR, Nadalutti C, Collighan R, Mongeot A, Griffin M, Mäki M, Kaukinen K, Lindfors K (2009) Celiac disease IgA modulates vascular permeability in vitro through the activity of transglutaminase 2 and RhoA. Cell Mol Life Sci 20:3375–3385
Cooke WT, Holmes GK (1984) Celiac disease. Churchill Livingstone, New York, pp 27–28
Aleanzi M, Demonte AM, Esper C, Garcilazo S, Waggener M (2001) Celiac disease: antibody recognition against native and selectively deamidated gliadin peptides. Clin Chem 47:2023–2028
Sulkanen S, Halttunen T, Laurila K, Kolho KL, Korponay-Szabo IR, Sarnesto A, Savilahti E, Collin P, Mäki M (1998) Tissue transglutaminase autoantibody enzyme-linked immunosorbent assay in detecting celiac disease. Gastroenterology 115:1322–1328
Dieterich W, Laag E, Schöpper H, Volta U, Ferguson A, Gillett H, Riecken EO, Schuppan D (1998) Autoantibodies to tissue transglutaminase as predictors of celiac disease. Gastroenterology 115:1317–1321
Klöck C, Khosla C (2012) Regulation of the activities of the mammalian transglutaminase family of enzymes. Protein Sci 12:1781–1791
Molberg O, Mcadam SN, Körner R, Quarsten H, Kristiansen C, Madsen L, Fugger L, Scott H, Norén O, Roepstorff P, Lundin KE, Sjöström H, Sollid LM (1998) Tissue transglutaminase selectively modifies gliadin peptides that are recognized by gut-derived T cells in celiac disease. Nat Med 4:713–717
Aeschlimann D, Thomazy V (2000) Protein crosslinking in assembly and remodelling of extracellular matrices: the role of transglutaminases. Connect Tissue Res 41:1–27
Akimov SS, Krylov D, Fleischman LF, Belkin AM (2000) Tissue transglutaminase is an integrin-binding adhesion co receptor for fibronectin. J Cell Biol 148:825–838
Wang Z, Collighan RJ, Pytel K, Rathbone DL, Li X, Griffin M (2012) Characterization of heparin-binding site of tissue transglutaminase: its importance in cell surface targeting, matrix deposition, and cell signaling. J Biol Chem 287:13063–13083
Kotsakis P, Griffin M (2007) Tissue transglutaminase in tumour progression: friend or foe? Amino Acids 33:373–384
Nadalutti C, Viiri KM, Kaukinen K, Mäki M, Lindfors K (2011) Extracellular transglutaminase 2 has a role in cell adhesion, whereas intracellular transglutaminase 2 is involved in regulation of endothelial cell proliferation and apoptosis. Cell Prolif 44:49–58
Haroon ZA, Hettash JM, Lai TS, Dewhirst MW, Greenberg CS (1999) Tissue transglutaminase is expressed, active and directly involved in rat dermal wound healing and angiogenesis. FASEB J 13:1787–1795
Jones RA, Kotsakis P, Johnson TS, Chau DY, Ali S, Melino G, Griffin M (2006) Matrix changes induced by transglutaminase 2 lead to inhibition of angiogenesis and tumor growth. Cell Death Differ 13:1442–1453
Munezane T, Hasegawa T, Suritala, Tanaka A, Okada K, Okita Y (2010) Activation of transglutaminase type 2 for aortic wall protection in a rat abdominal aortic aneurysm formation. J Vasc Surg 52:967–974
Jones RA, Wang Z, Dookie S, Griffin M (2013) The role of TG2 in ECV304-related vasculogenic. Amino Acids 44:89–101
Myrsky E, Kaukinen K, Syrjanen M, Korponay-Szabò IR, Maki M, Lindfors K (2008) Coeliac disease-specific autoantibodies targeted against transglutaminase 2 disturb angiogenesis. Clin Exp Immunol 152:111–119
Caja S, Myrsky E, Korponay-Szabo IR, Nadalutti C, Sulic AM, Lavric M, Sblattero D, Marzari R, Collighan R, Mongeot A, Griffin M, Mäki M, Kaukinen K, Lindfors K (2010) Inhibition of transglutaminase 2 enzymatic activity ameliorates the anti-angiogenic effects of coeliac disease autoantibodies. Scand J Gastroenterol 45:421–427
Martucciello S, Lavric M, Toth B, Korponay-Szabo I, Nadalutti C, Myrsky E, Rauhavirta T, Esposito C, Sulic AM, Sblattero D, Marzari R, Mäki M, Kaukinen K, Lindfors K, Caja S (2012) RhoB is associated with the anti-angiogenic effects of celiac patient transglutaminase 2-targeted autoantibodies. J Mol Med 90:817–826
Griffin M, Mongeot A, Collighan R, Saint RE, Jones RA, Coutts IG, Rathbone DL (2008) Synthesis of potent water-soluble tissue transglutaminase inhibitors. Bioorg Med Chem Lett 18:5559–5562
Forsprecher J, Wang Z, Nelea V, Kaartinen MT (2009) Enhanced osteoblast adhesion on transglutaminase 2-crosslinked fibronectin. Amino Acids 36:747–753
Tomar A, Lim ST, Lim Y, Schlaepfer DD (2009) A FAK-p120RasGAP-p190RhoGAP complex regulates polarity in migrating cells. Cell Sci 122:3005
Zemskov EA, Janiak A, Hang J, Waghray A, Belkin AM (2006) The role of tissue transglutaminase in cell-matrix interactions. Front Biosci 11:1057–1076
Scarpellini A, Germack R, Lortat-Jacob H, Muramatsu T, Billett E, Johnson T, Verderio EAM (2009) Heparan sulfate proteoglycans are receptors for the cell-surface trafficking and biological activity of transglutaminase-2. J Biol Chem 284:18411–18423
Belkin AM, Akimov SS, Zaritskaya LS, Ratnikov BI, Deryugina EI, Strongin AY (2001) Matrix-dependent proteolysis of surface transglutaminase by membrane-type metalloproteinase regulates cancer cell adhesion and locomotion. J Biol Chem 276:18415–18422
Belkin A (2011) Extracellular TG2: emerging functions and regulation. FEBS J 278:4704–4716
Demali KA (2004) Vinculin-a dynamic regulator of cell adhesion. Trends Biochem Sci 11:565–567
Midwood KS, Williams LV, Schwarzbauer JE (2004) Tissue repair and the dynamics of the extracellular matrix. Int J Biochem Cell Biol 36:1031–1037
Al-Jallad HF, Nakano Y, Chen JL, McMillan E, Lefebvre C, Kaartinen MT (2006) Transglutaminase activity regulates osteoblast differentiation and matrix mineralization in MC3T3-E1 osteoblast cultures. Matrix Biol 25:135–148
Iversen R, Di Niro R, Stamnaes J, Lundin KE, Wilson PC, Sollid LM (2013) Transglutaminase 2-specific autoantibodies in celiac disease target clustered, N-terminal epitopes not displayed on the surface of cells. J Immunol 190:5981–5991
Teesalu K, Panarina M, Uibo O, Uibo R, Utt M (2012) Autoantibodies from patients with celiac disease inhibit transglutaminase 2 binding to heparin/heparan sulfate and interfere with intestinal epithelial cell adhesion. Amino Acids 42:1055–1064
Mesin L, Sollid LM, Di Niro R (2012) The intestinal B-cell response in celiac disease. Front Immunol 3:313
Janiak A, Zemskov EA, Belkin AM (2006) Cell surface transglutaminase promotes RhoA activation via integrin clustering and suppression of the Src-p190RhoGAP signaling pathway. Mol Biol Cell 17:1606–1619
Telci D, Wang Z, Li X, Verderio EA, Humphries MJ, Baccarini M, Basaga H, Griffin M (2008) Fibronectin-tissue transglutaminase matrix rescues RGD-impaired cell adhesion through syndecan-4 and beta1 integrin co-signaling. J Biol Chem 283:20937–20947
Shaoul R, Lerner A (2007) Associated autoantibodies in celiac disease. Autoimmun Rev 8:559–565
Myrsky E, Syrjänen M, Korponay-Szabo IR, Mäki M, Kaukinen K, Lindfors K (2009) Altered small-bowel mucosal vascular network in untreated coeliac disease. Scand J Gastroenterol 44:162–167
Acknowledgments
We thank Kaisa Teittinen for comments on the manuscript. The Coeliac Disease Study Group was financially supported by the Academy of Finland, the Tampere Graduate Program in Biomedicine and Biotechnology, the Sigrid Juselius Foundation, the Research Fund of the Finnish Coeliac Society, the Competitive State Research Financing of the Expert Responsibility Area of Tampere University Hospital (grant numbers 9P020 and 9P033) and the Pediatric Research Foundation, Elna Kaarina Savolainen’s fund allocated for the development of cancer treatment and the European Commission IAPP grant TRANSCOM (Contract number PIA-GA-2010-251506). Further, the grants OTKA K101788, NK105046 and TÁMOP 4.2.2.11/1/KONV-2012-0023 are also acknowledged.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nadalutti, C.A., Korponay-Szabo, I.R., Kaukinen, K. et al. Celiac disease patient IgA antibodies induce endothelial adhesion and cell polarization defects via extracellular transglutaminase 2. Cell. Mol. Life Sci. 71, 1315–1326 (2014). https://doi.org/10.1007/s00018-013-1455-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00018-013-1455-5