Reversible opening of the blood-brain barrier by claudin-5-binding variants of Clostridium perfringens enterotoxin's claudin-binding domain

• Verfasst von: Neuhaus, Winfried [VerfasserIn]
• Verfasst von: Mahringer, Anne [VerfasserIn]
• Titel: Reversible opening of the blood-brain barrier by claudin-5-binding variants of Clostridium perfringens enterotoxin's claudin-binding domain
• Verf.angabe: Winfried Neuhaus, Anna Piontek, Jonas Protze, Miriam Eichner, Anne Mahringer, Eva-Anne Subileau, In-Fah M. Lee, Jörg D. Schulzke, Gerd Krause, Jörg Piontek
• E-Jahr: 2018
• Jahr: 2 February 2018
• Umfang: 15 S.
• Fussnoten: Gesehen am 19.02.2020
• Titel Quelle: Enthalten in: Biomaterials
• Ort Quelle: Amsterdam [u.a.] : Elsevier Science, 1980
• Jahr Quelle: 2018
• Band/Heft Quelle: 161(2018), Seite 129-143
• ISSN Quelle: 0142-9612
• DOI: doi:10.1016/j.biomaterials.2018.01.028
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Blood brain barrier
• Sach-SW: Claudin
• Sach-SW: enterotoxin
• Sach-SW: Paracellular drug delivery
• Sach-SW: Tight junction
• K10plus-PPN: 169038638X

Abstract

The blood-brain barrier (BBB) prevents entry of neurotoxic substances but also that of drugs into the brain. Here, the paracellular barrier is formed by tight junctions (TJs) with claudin-5 (Cldn5) being the main sealing constituent. Transient BBB opening by targeting Cldn5 could improve paracellular drug delivery. The non-toxic C-terminal domain of Clostridium perfringens enterotoxin (cCPE) binds to a subset of claudins, e.g., Cldn3, -4. Structure-based mutagenesis was used to generate Cldn5-binding variants (cCPE-Y306W/S313H and cCPE-N218Q/Y306W/S313H). These cCPE-variants were tested for transient TJ opening using multiple in vitro BBB models: Primary porcine brain endothelial cells, coculture of primary rat brain endothelial cells with astrocytes and mouse cerebEND cells. cCPE-Y306W/S313H and cCPE-N218Q/Y306W/S313H but neither cCPE-wt nor cCPE-Y306A/L315A (not binding to claudins) decreased transendothelial electrical resistance in a concentration-dependent and reversible manner. Furthermore, permeability of carboxyfluorescein (with size of CNS drugs) was increased. cCPE-Y306W/S313H but neither cCPE-wt nor cCPE-Y306A/L315A bound to Cldn5-expressing brain endothelial cells. However, freeze-fracture EM showed that cCPE-Y306W/S313H did not cause drastic TJ breakdown. In sum, Cldn5-binding cCPE-variants enabled mild and transient opening of brain endothelial TJs. Using reliable in vitro BBB models, the results demonstrate that cCPE-based biologics designed to bind Cldn5 improve paracellular drug delivery across the BBB.