{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T17:10:57Z","timestamp":1785431457737,"version":"3.56.0"},"posted":{"date-parts":[[2026,7,20]]},"group-title":"Physiology","reference-count":70,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T00:00:00Z","timestamp":1784505600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["DK114024"],"award-info":[{"award-number":["DK114024"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,20]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Defective intestinal epithelial tight junction (TJ) barrier function and endoplasmic reticulum (ER) stress are central pathological features of inflammatory bowel disease (IBD), yet the molecular mechanisms ER stress to TJ disruption remains poorly understood. Here, we investigated the role of autophagy in regulating intestinal TJ homeostasis during ER stress. ER stress was elevated in inflamed Crohn\u2019s disease tissue and chronic dextran sulfate sodium (DSS) colitis. In human intestinal epithelial Caco-2 monolayers, murine colon, and human colonic explants, induction of ER stress with tunicamycin, thapsigargin, or brefeldin A disrupted TJ barrier integrity, as demonstrated by reduced transepithelial electrical resistance and increased paracellular permeability. ER stress selectively increased the pore-forming TJ protein claudin-2 and altered occludin localization without significantly affecting other claudins. Pharmacologic activation of autophagy with rapamycin attenuated ER stress, restored TJ barrier function, reduced claudin-2 accumulation, and preserved occludin localization. Conversely, CRISPR-Cas9-mediated deletion of autophagy gene ATG7 exacerbated ER stress, apoptosis, and TJ barrier dysfunction in vitro, while intestinal epithelial-specific Atg7 knockout mice exhibited enhanced ER stress-induced intestinal permeability in-vivo. Mechanistically, prolonged ER stress impaired autophagic flux through IRE1\u03b1 kinase signaling, resulting in accumulation of p62 and claudin-2. Inhibition of IRE1\u03b1 kinase activity restored autophagy, reduced claudin-2 levels, and preserved TJ barrier function. We further identified adaptor-associated kinase 1 (AAK1) as a downstream mediator of IRE1\u03b1 signaling during ER stress, with increased AP2M1 phosphorylation and altered claudin-2 trafficking. Claudin-2 overexpression alone induced ER stress and lysosomal damage, suggesting a feed-forward mechanism amplifying epithelial injury. Finally, enteric rapamycin administration reduced ER stress and restored autophagy in murine DSS colitis. Collectively, these findings identify an IRE1\u03b1\u2013AAK1\u2013autophagy axis as a critical regulator of intestinal TJ barrier integrity during ER stress.<\/jats:p>","DOI":"10.64898\/2026.07.15.738672","type":"posted-content","created":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T04:20:13Z","timestamp":1784607613000},"source":"Crossref","is-referenced-by-count":0,"title":["Autophagy prevents ER stress-induced Tight Junction barrier disruption via claudin-2 homeostasis"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4260-0381","authenticated-orcid":false,"given":"Priya","family":"Arumugam","sequence":"first","affiliation":[{"name":"Division of Gastroenterology and Hepatology, Department of Medicine, Penn State College of Medicine, Hershey, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2692-2532","authenticated-orcid":false,"given":"Kushal","family":"Saha","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology and Hepatology, Department of Medicine, Penn State College of Medicine, Hershey, PA, USA"},{"name":"Division of Gastroenterology, Department of Medicine, Icahn School of Medicine, Mount Sinai Hospital, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashwinkumar","family":"Subramenium Ganapathy","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology and Hepatology, Department of Medicine, Penn State College of Medicine, Hershey, PA, USA"},{"name":"Syngene International PV Ltd, Bangalore, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandra","family":"Wang","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology and Hepatology, Department of Medicine, Penn State College of Medicine, Hershey, PA, USA"},{"name":"Johnson & Johnson PV Ltd, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leonard","family":"Harris","sequence":"additional","affiliation":[{"name":"Division of Colon and Rectal Surgery, Department of Surgery, Penn State College of Medicine, Hershey, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1211-3040","authenticated-orcid":false,"given":"Gregory","family":"Yochum","sequence":"additional","affiliation":[{"name":"Division of Colon and Rectal Surgery, Department of Surgery, Penn State College of Medicine, Hershey, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2368-7290","authenticated-orcid":false,"given":"Prashant","family":"Nighot","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology and Hepatology, Department of Medicine, Penn State College of Medicine, Hershey, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026072304250889000_2026.07.15.738672v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/361552a0"},{"key":"2026072304250889000_2026.07.15.738672v1.2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01213309"},{"key":"2026072304250889000_2026.07.15.738672v1.3","unstructured":"Burden of digestive diseases in the United States, 2008. 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