{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,29]],"date-time":"2025-10-29T03:10:07Z","timestamp":1761707407752,"version":"build-2065373602"},"reference-count":40,"publisher":"Wiley","issue":"18","license":[{"start":{"date-parts":[[2025,9,24]],"date-time":"2025-09-24T00:00:00Z","timestamp":1758672000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004479","name":"Natural Science Foundation of Jiangxi Province","doi-asserted-by":"publisher","award":["20232BAB206001"],"award-info":[{"award-number":["20232BAB206001"]}],"id":[{"id":"10.13039\/501100004479","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82360016"],"award-info":[{"award-number":["82360016"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["faseb.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FASEB Journal"],"published-print":{"date-parts":[[2025,9,30]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Pulmonary toxicity is a common adverse effect of temsirolimus (a first\u2010generation mammalian target of rapamycin (mTOR) inhibitor), but its mechanisms are unclear. Disruption of endothelial\u2010barrier integrity is key in the pathophysiology of lung injury. We investigated the role and mechanisms of endothelial\u2010barrier dysfunction in the pathogenesis of temsirolimus\u2010induced lung injury. We evaluated the impact of temsirolimus on the permeability of human pulmonary microvascular endothelial cells (HPMECs) using transendothelial electrical resistance and albumin leakage while simultaneously investigating its effects on Ca\n                    <jats:sup>2+<\/jats:sup>\n                    release via ryanodine receptors (RyRs) in endothelial cells (ECs). The roles of myosin light chain kinase (MLCK) in endothelial\u2010barrier permeability were studied in MLCK small interfering (si)RNA\u2010transfected HPMECs. In addition, we established a mouse model subjected to intraperitoneal injections of temsirolimus to explore its effects on pulmonary vascular permeability and lung injury. We highlighted the contribution of the MLCK to temsirolimus\u2010induced vascular hyperpermeability and lung injury, supported by studies in two lines of transgenic mice with knocking out MLCK or conditional deletion of MLCK in the endothelium. Temsirolimus increased the permeability of HPMECs, which was correlated with the phosphorylation of myosin light chain (MLC), MLCK activation, and the formation of F\u2010actin stress fibers. Temsirolimus caused a rise in intracellular Ca\n                    <jats:sup>2+<\/jats:sup>\n                    leakage within HPMECs, an effect that was reversed by pretreatment with ryanodine. The latter diminished the phosphorylation of MLCK\/MLC induced by temsirolimus, which subsequently led to disruption of the endothelial barrier in HPMECs. Aligning with these in\u00a0vitro findings, temsirolimus administration resulted in dysfunction of the lung\u2013vascular barrier, characterized by increased protein levels in bronchoalveolar lavage fluid (BALF) and increased permeability of the lung capillary endothelium. Mice with systemic and EC\u2010specific MLCK knockout exhibited reduced temsirolimus\u2010induced pulmonary microvascular hyperpermeability and lung injury. Temsirolimus induced pulmonary endothelial hyperpermeability mediated (at least in part) by the Ca\n                    <jats:sup>2+<\/jats:sup>\n                    \u2010dependent MLCK\/p\u2010MLC pathway caused EC contraction and contributed to lung injury through mTOR\u2010independent mechanisms.\n                  <\/jats:p>","DOI":"10.1096\/fj.202501447r","type":"journal-article","created":{"date-parts":[[2025,9,24]],"date-time":"2025-09-24T15:32:19Z","timestamp":1758727939000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Myosin Light Chain Kinase\u2013Mediated Endothelial Hyperpermeability Underlies Temsirolimus\u2010Induced Lung Injury"],"prefix":"10.1096","volume":"39","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6349-4341","authenticated-orcid":false,"given":"Xiaolin","family":"Chen","sequence":"first","affiliation":[{"name":"Department of Clinical Laboratory, Pingxiang People's Hospital Gannan Medical University  Pingxiang Jiangxi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jianhui","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, Pingxiang People's Hospital Gannan Medical University  Pingxiang Jiangxi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuihong","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, Pingxiang People's Hospital Gannan Medical University  Pingxiang Jiangxi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Danjing","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, Pingxiang People's Hospital Gannan Medical University  Pingxiang Jiangxi China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,9,24]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.03.035"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1716173114"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI64099"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1097\/TP.0000000000004336"},{"key":"e_1_2_12_6_1","article-title":"Target of Rapamycin Inhibitors (TOR\u2010I; Sirolimus and Everolimus) for Primary Immunosuppression in Kidney Transplant Recipients","volume":"12","author":"Hahn D.","year":"2019","journal-title":"Cochrane Database of Systematic 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