| dc.contributor.author | Stricker, Sebastian | |
| dc.contributor.author | de Laffolie, Jan | |
| dc.contributor.author | Zimmer, Klaus-Peter | |
| dc.contributor.author | Rudloff, Silvia | |
| dc.date.accessioned | 2023-04-17T06:31:12Z | |
| dc.date.available | 2023-04-17T06:31:12Z | |
| dc.date.issued | 2023 | |
| dc.identifier.uri | https://jlupub.ub.uni-giessen.de//handle/jlupub/16229 | |
| dc.identifier.uri | http://dx.doi.org/10.22029/jlupub-15612 | |
| dc.description.abstract | Enzymatic modification of gliadin peptides by human transglutaminase 2 (TG2) is a key mechanism in the pathogenesis of celiac disease (CD) and represents a potential therapeutic target. Recently, we have identified the small oxidative molecule PX-12 as an effective inhibitor of TG2 in vitro. In this study, we further investigated the effect of PX-12 and the established active-site directed inhibitor ERW1041 on TG2 activity and epithelial transport of gliadin peptides. We analyzed TG2 activity using immobilized TG2, Caco-2 cell lysates, confluent Caco-2 cell monolayers and duodenal biopsies from CD patients. TG2-mediated cross-linking of pepsin-/trypsin-digested gliadin (PTG) and 5BP (5-biotinamidopentylamine) was quantified by colorimetry, fluorometry and confocal microscopy. Cell viability was tested with a resazurin-based fluorometric assay. Epithelial transport of promofluor-conjugated gliadin peptides P31-43 and P56-88 was analyzed by fluorometry and confocal microscopy. PX-12 reduced TG2-mediated cross-linking of PTG and was significantly more effective than ERW1041 (10 µM, 15 ± 3 vs. 48 ± 8%, p < 0.001). In addition, PX-12 inhibited TG2 in cell lysates obtained from Caco-2 cells more than ERW1041 (10 µM; 12 ± 7% vs. 45 ± 19%, p < 0.05). Both substances inhibited TG2 comparably in the intestinal lamina propria of duodenal biopsies (100 µM, 25 ± 13% vs. 22 ± 11%). However, PX-12 did not inhibit TG2 in confluent Caco-2 cells, whereas ERW1041 showed a dose-dependent effect. Similarly, epithelial transport of P56-88 was inhibited by ERW1041, but not by PX-12. Cell viability was not negatively affected by either substance at concentrations up to 100 µM. PX-12 did not reduce TG2 activity or gliadin peptide transport in confluent Caco-2 cells. This could be caused by rapid inactivation or degradation of the substance in the Caco-2 cell culture. Still, our in vitro data underline the potential of the oxidative inhibition of TG2. The fact that the TG2-specific inhibitor ERW1041 reduced the epithelial uptake of P56-88 in Caco-2 cells further strengthens the therapeutic potential of TG2 inhibitors in CD. | |
| dc.description.sponsorship | Deutsche Forschungsgemeinschaft (DFG); ROR-ID:018mejw64 | |
| dc.language.iso | en | |
| dc.rights | Namensnennung 4.0 International | |
| dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | |
| dc.subject | celiac disease | |
| dc.subject | transglutaminase 2 | |
| dc.subject | TG2 inhibitor | |
| dc.subject | PX-12 | |
| dc.subject | gliadin transport | |
| dc.subject.ddc | ddc:610 | |
| dc.title | Inhibition of Transglutaminase 2 as a Therapeutic Strategy in Celiac Disease - In Vitro Studies in Intestinal Cells and Duodenal Biopsies | |
| dc.type | article | |
| local.affiliation | FB 11 - Medizin | |
| local.project | GU 405/14-1 | |
| local.source.spage | 1 | |
| local.source.epage | 14 | |
| local.source.journaltitle | International journal of molecular sciences | |
| local.source.volume | 24 | |
| local.source.articlenumber | 4795 | |
| local.source.uri | https://doi.org/10.3390/ijms24054795 | |
