{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T22:46:06Z","timestamp":1781736366406,"version":"3.54.5"},"reference-count":69,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2024,4,17]],"date-time":"2024-04-17T00:00:00Z","timestamp":1713312000000},"content-version":"vor","delay-in-days":16,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Clinical &amp;amp; Translational Med"],"published-print":{"date-parts":[[2024,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Recurrent malignant pleural effusion (MPE) resulting from non\u2010small\u2010cell lung cancer (NSCLC) is easily refractory to conventional therapeutics and lacks predictive markers. The cellular or genetic signatures of recurrent MPE still remain largely uncertain.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>16 NSCLC patients with pleural effusions were recruited, followed by corresponding treatments based on primary tumours. Non\u2010recurrent or recurrent MPE was determined after 3\u20136 weeks of treatments. The status of MPE was verified by computer tomography (CT) and cytopathology, and the baseline pleural fluids were collected for single\u2010cell RNA sequencing (scRNA\u2010seq). Samples were then integrated and profiled. Cellular communications and trajectories were inferred by bioinformatic algorithms. Comparative analysis was conducted and the results were further validated by quantitative polymerase chain reaction (qPCR) in a larger MPE cohort from the authors' centre (<jats:italic>n<\/jats:italic>\u00a0=\u00a064).<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The scRNA\u2010seq revealed that 33\u00a0590 cells were annotated as 7 major cell types and further characterized into 14 cell clusters precisely. The cell cluster C1, classified as Epithelial Cell Adhesion Molecule (<jats:italic>EpCAM<\/jats:italic>)+ metastatic cancer cell and correlated with activation of tight junction and adherence junction, was significantly enriched in the recurrent MPE group, in which Claudin\u20104 (<jats:italic>CLDN4<\/jats:italic>) was identified. The subset cell cluster C3 of C1, which was enriched in recurrent MPE and demonstrated a phenotype of ameboidal\u2010type cell migration, also showed a markedly higher expression of <jats:italic>CLDN4<\/jats:italic>. Meanwhile, the expression of <jats:italic>CLDN4<\/jats:italic> was positively correlated with E74 Like ETS Transcription Factor 3 (<jats:italic>ELF3<\/jats:italic>), <jats:italic>EpCAM<\/jats:italic> and Tumour Associated Calcium Signal Transducer 2 (<jats:italic>TACSTD2<\/jats:italic>), independent of driver\u2010gene status. <jats:italic>CLDN4<\/jats:italic> was also found to be associated with the expression of Hypoxia Inducible Factor 1 Subunit Alpha (<jats:italic>HIF1A<\/jats:italic>) and Vascular Endothelial Growth Factor A (<jats:italic>VEGFA<\/jats:italic>), and the cell cluster C1 was the major mediator in cellular communication of <jats:italic>VEGFA<\/jats:italic> signalling. In the extensive MPE cohort, a notably increased expression of <jats:italic>CLDN4<\/jats:italic> in cells from pleural effusion among patients diagnosed with recurrent MPE was observed, compared with the non\u2010recurrent group, which was also associated with a trend towards worse overall survival (OS).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p><jats:italic>CLDN4<\/jats:italic> could be considered as a predictive marker of recurrent MPE among patients with advanced NSCLC. Further validation for its clinical value in cohorts with larger sample size and in\u2010depth mechanism studies on its biological function are warranted.<\/jats:p><\/jats:sec><jats:sec><jats:title>Trial registration<\/jats:title><jats:p>Not applicable.<\/jats:p><\/jats:sec>","DOI":"10.1002\/ctm2.1649","type":"journal-article","created":{"date-parts":[[2024,4,17]],"date-time":"2024-04-17T11:28:29Z","timestamp":1713353309000},"source":"Crossref","is-referenced-by-count":15,"title":["Single\u2010cell transcriptomics reveal metastatic <i>CLDN4<\/i>+ cancer cells underlying the recurrence of malignant pleural effusion in patients with advanced non\u2010small\u2010cell lung cancer"],"prefix":"10.1002","volume":"14","author":[{"given":"Xiaoshen","family":"Zhang","sequence":"first","affiliation":[{"name":"School of Medicine Tongji University Shanghai China"},{"name":"Department of Medical Oncology Shanghai Pulmonary Hospital, Tongji University School of Medicine Shanghai 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