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Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2025,6,17]]},"abstract":"<jats:p>\n                    The \u03b1-hemolysin (HlyA) of uropathogenic\n                    <jats:italic toggle=\"yes\">Escherichia coli<\/jats:italic>\n                    (UPEC) is a pore-forming toxin (PFT) that is thought to function by disrupting the host cell plasma membrane. Although CD18 (LFA-1) has been implicated as a receptor on myeloid cells, the mechanisms underlying HlyA cytotoxicity to epithelial cells are poorly defined. Here, we show that HlyA secretion by UPEC markedly intensifies renal tubular epithelial injury in a murine model of ascending pyelonephritis. A CRISPR-Cas9 loss-of-function screen in renal collecting duct cells revealed an unexpected requirement for clathrin-mediated endocytosis in HlyA-induced cytotoxicity. Following internalization, HlyA triggered lysosomal permeabilization, resulting in protease leakage, cytoplasmic acidification, and mitochondrial impairment, culminating in rapid epithelial cell death\u2014a pathway distinct from canonical membrane-disrupting mechanisms of other PFTs. Moreover, we identify the low-density lipoprotein receptor (LDLR) as a critical epithelial receptor for HlyA; genetic ablation or competitive inhibition of the HlyA\u2013LDLR interaction fully abrogated cytotoxicity. Our findings detail a paradigm for HlyA function in which epithelial toxicity relies on LDLR-mediated endocytic uptake rather than plasma membrane poration. These mechanistic insights illuminate potential therapeutic strategies to attenuate HlyA-mediated tissue damage during UPEC infections.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2505482122","type":"journal-article","created":{"date-parts":[[2025,6,12]],"date-time":"2025-06-12T10:49:07Z","timestamp":1749725347000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":6,"title":["LDL receptor\u2013mediated endocytosis of\n                    <i>Escherichia coli<\/i>\n                    \u03b1-hemolysin mediates renal epithelial toxicity"],"prefix":"10.1073","volume":"122","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7876-3521","authenticated-orcid":false,"given":"Hunter W.","family":"Kuhn","sequence":"first","affiliation":[{"name":"Department of Pediatrics, Washington University School of Medicine","place":["St. 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