Identification of claudin-4 binder that attenuates tight junction barrier function by TR-FRET-based screening assay. Issue 1 (December 2017)

• Record Type: Journal Article
• Authors: Watari, Akihiro; Kodaka, Miki; Matsuhisa, Koji; Sakamoto, Yuta; Hisaie, Kota; Kawashita, Norihito; Takagi, Tatsuya; Yamagishi, Yoshiaki; Suzuki, Hidehiko; Tsujino, Hirofumi; Yagi, Kiyohito; Kondoh, Masuo
• Is Part Of: Scientific reports. Volume 7:Issue 1(2017)
• Page Start: 1
• Page End: 12
• Publication Date: 2017-12
• Subjects: Natural history -- Research -- Periodicals; Biology -- Research -- Periodicals; Physical sciences -- Research -- Periodicals; Earth sciences -- Research -- Periodicals; Environmental sciences -- Research -- Periodicals; 502.85
• DOI: 10.1038/s41598-017-15108-y
• Languages: English
• ISSNs: 2045-2322

Abstract

Abstract Claudins are key functional and structural components of tight junctions (TJs) in epithelial cell sheets. The C-terminal fragment ofClostridium perfringens enterotoxin (C-CPE) binds to claudin-4 and reversibly modulates intestinal TJ seals, thereby enhancing paracellular transport of solutes. However, the use of C-CPE as an absorption enhancer is limited by the molecule's immunogenicity and manufacturing cost. Here, we developed a high-throughput screening system based on the Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) method to identify claudin-4 binders in a library collection of 32, 560 compounds. Thiostrepton, identified from the screen, decreased transepithelial electrical resistance and increased flux of 4-kDa fluorescein isothiocyanate–labelled dextran (FD-4) in Caco-2 cell monolayers, a model of intestinal epithelium. Thiostrepton changed the expression, but not the localisation, of TJ components. Treatment of rat jejunum with thiostrepton increased the absorption of FD-4 without tissue toxicity, indicating that thiostrepton is a novel claudin-4 binder that enhances intestinal permeability. The screening system may therefore be a useful tool for identifying claudin-4 binders to enhance drug absorption in mucosa.