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 transfectants. 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 assessed 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 N(146) residue in hCldn-14 and the S(151) 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 reportedly expressed in mammalian intestines, the current results support the possibility that these two hCldns contribute to natural CPE-mediated
gastrointestinal disease and could be CPE-based therapeutic targets for
cancers overexpressing those
claudins. 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.
IMPORTANCE:
Clostridium perfringens enterotoxin (CPE) is responsible for the gastrointestinal symptoms of the second-most-common bacterial
food-borne illness and is also being explored for use as a
cancer therapeutic or for increasing
drug delivery. Until now, the only known human CPE receptors were
claudin-3 and -4. This work shows that human
claudin-8 and -14 can also bind CPE and convey cytotoxicity, although slightly less efficiently than
claudin-3 and -4. The
claudin-8 and -14 residues responsible for this weaker CPE binding were identified, shedding new light on CPE:
claudin interactions.