Claudin targeting as an effective tool for directed barrier modulation of the viable epidermis.

Laura-Sophie Beier, Ayk Waldow, Saeed Khomeijani Farahani, Roman Mannweiler, Sabine Vidal-Y-Sy, Johanna M Brandner, Jörg Piontek, Dorothee Günzel

Annals of the New York Academy of Sciences 2022 Nov

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Tight junction (TJ) formation is vital for epidermal barrier function. We aimed to specifically manipulate TJ barriers in the reconstructed human epidermis (RHE) by claudin-1 and -4 knockdown (KD) and by claudin-binding fusion proteins of glutathione S-transferase and modified C-terminal fragments of Clostridium perfringens enterotoxin (GST-cCPE). Impedance spectroscopy and tracer permeability imaging were employed for functional barrier assessment and investigation of claudin contribution. KD of claudin-1, but not claudin-4, impaired the paracellular barrier in vitro. Similarly, claudin-binding GST-cCPE variants weakened the paracellular but not the stratum corneum barrier. Combining both TJ targeting methods, we found that claudin-1 targeting by GST-cCPE after claudin-4 KD led to a marked decrease in paracellular barrier properties. Conversely, after claudin-1 KD, GST-cCPE did not further impair the barrier. Comparison of GST-cCPE variants with different claudin-1/claudin-4 affinities, NHS-fluorescein tracer detection, and immunostaining of RHE paraffin sections showed that GST-cCPE variants bind to extrajunctional claudin-1 and -4, which are differentially distributed along the stratum basale-stratum granulosum axis. GST-cCPE binding blocks these claudins, thereby specifically opening the paracellular barrier of RHE. The data indicate a critical role for claudin-1 in regulating paracellular permeability for ions and small molecules in the viable epidermis. Claudin targeting is presented as a proof-of-concept for precise barrier modulation. © 2022 The Authors. Annals of the New York Academy of Sciences published by Wiley Periodicals LLC on behalf of New York Academy of Sciences.

Citation

Laura-Sophie Beier, Ayk Waldow, Saeed Khomeijani Farahani, Roman Mannweiler, Sabine Vidal-Y-Sy, Johanna M Brandner, Jörg Piontek, Dorothee Günzel. Claudin targeting as an effective tool for directed barrier modulation of the viable epidermis. Annals of the New York Academy of Sciences. 2022 Nov;1517(1):251-265