Human Claudin-8 and-14 Are Receptors Capable of Conveying the Cytotoxic Effects of Clostridium perfringens Enterotoxin

ABSTRACT Clostridium perfringens enterotoxin (CPE) contributes to several important human gastrointestinal (GI) diseases. This toxin and its derivatives are also being explored for translational applications, i.e., cancer therapy or drug delivery. Some, but not all, members of the 24-member claudin (Cldn) family of mammalian tight junction proteins can serve as CPE receptors. Among the human Cldns (hCldns), hCldn-3 and-4 are known to convey CPE sensitivity when expressed by fibroblast transfec-tants. However, other Cldns are also reportedly expressed in the intestines, where they might contribute to natural CPE-mediated GI disease, and in other organs, where they might react with CPE-based therapeutics. Therefore, the current study as-sessed whether two additional hCldns beside hCldn-3 and-4 are also functional CPE receptors. Using Cldn-expressing transfectants, hCldn-8 and-14 were shown to convey CPE-mediated cytotoxicity at pathophysiologically relevant concentrations of this toxin, although ~2-to-10-fold less efficiently than hCldn-4. Site-directed mutagenesis then demonstrated that the N146 residue in hCldn-14 and the S151 residue in hCldn-8 are largely responsible for modulating the weaker CPE binding properties of hCldn-8 and-14 versus hCldn-4, which broadens understanding of Cldn:CPE binding interactions. Since Cldn-8 and-14 are re-portedly expressed in mammalian intestines, the current results support the possibility that these two hCldns contribute to nat-ural CPE-mediated gastrointestinal disease and could be CPE-based therapeutic targets for cancers overexpressing those clau-dins. However, these results also suggest caution during therapeutic use of CPE, which might trigger toxic side effects in normal human tissues producing hCldn-8 or-14, as well as in those producing hCldn-3 or-4