TY  - JOUR
AU  - Waldow, Ayk
AU  - Beier, Laura-Sophie
AU  - Arndt, Janine
AU  - Schallenberg, Simon
AU  - Vollbrecht, Claudia
AU  - Bischoff, Philip
AU  - Farrera-Sal, Martí
AU  - Loch, Florian N
AU  - Bojarski, Christian
AU  - Schumann, Michael
AU  - Winkler, Lars
AU  - Kamphues, Carsten
AU  - Ehlen, Lukas
AU  - Piontek, Jörg
TI  - cCPE Fusion Proteins as Molecular Probes to Detect Claudins and Tight Junction Dysregulation in Gastrointestinal Cell Lines, Tissue Explants and Patient-Derived Organoids.
JO  - Pharmaceutics
VL  - 15
IS  - 7
SN  - 1999-4923
CY  - Basel
PB  - MDPI
M1  - DKFZ-2023-01537
SP  - 1980
PY  - 2023
AB  - Claudins regulate paracellular permeability, contribute to epithelial polarization and are dysregulated during inflammation and carcinogenesis. Variants of the claudin-binding domain of Clostridium perfringens enterotoxin (cCPE) are highly sensitive protein ligands for generic detection of a broad spectrum of claudins. Here, we investigated the preferential binding of YFP- or GST-cCPE fusion proteins to non-junctional claudin molecules. Plate reader assays, flow cytometry and microscopy were used to assess the binding of YFP- or GST-cCPE to non-junctional claudins in multiple in vitro and ex vivo models of human and rat gastrointestinal epithelia and to monitor formation of a tight junction barrier. Furthermore, YFP-cCPE was used to probe expression, polar localization and dysregulation of claudins in patient-derived organoids generated from gastric dysplasia and gastric cancer. Live-cell imaging and immunocytochemistry revealed cell polarity and presence of tight junctions in glandular organoids (originating from intestinal-type gastric cancer and gastric dysplasia) and, in contrast, a disrupted diffusion barrier for granular organoids (originating from discohesive tumor areas). In sum, we report the use of cCPE fusion proteins as molecular probes to specifically and efficiently detect claudin expression, localization and tight junction dysregulation in cell lines, tissue explants and patient-derived organoids of the gastrointestinal tract.
KW  - adenoma (Other)
KW  - claudin (Other)
KW  - clostridium perfringens enterotoxin (Other)
KW  - colon (Other)
KW  - gastric cancer (Other)
KW  - gastric dysplasia (Other)
KW  - organoids (Other)
KW  - tight junction (Other)
KW  - tumor (Other)
LB  - PUB:(DE-HGF)16
C6  - pmid:37514167
C2  - pmc:PMC10385049
DO  - DOI:10.3390/pharmaceutics15071980
UR  - https://inrepo02.dkfz.de/record/277869
ER  -