{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T22:50:27Z","timestamp":1788648627555,"version":"build-2803163510"},"reference-count":43,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2022,12,16]],"date-time":"2022-12-16T00:00:00Z","timestamp":1671148800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000055","name":"National Institute on Deafness and Other Communication Disorders","doi-asserted-by":"publisher","award":["1R01DC018567"],"award-info":[{"award-number":["1R01DC018567"]}],"id":[{"id":"10.13039\/100000055","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000055","name":"National Institute on Deafness and Other Communication Disorders","doi-asserted-by":"publisher","award":["R21DC017225"],"award-info":[{"award-number":["R21DC017225"]}],"id":[{"id":"10.13039\/100000055","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Laryngoscope"],"published-print":{"date-parts":[[2023,9]]},"abstract":"<jats:sec><jats:title>Objectives<\/jats:title><jats:p>Idiopathic subglottic stenosis (iSGS) is characterized by progressive fibrosis and subglottic luminal narrowing. Currently, immune characterization has focused on T\u2010cells; however, macrophages remain largely unexplored. The goals of this study are to characterize the transcriptome of iSGS macrophages and the fibrogenic nature of identifed biomarkers.<\/jats:p><\/jats:sec><jats:sec><jats:title>Study Design<\/jats:title><jats:p>Bioinformatics and <jats:italic>in vitro<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Human tracheal biopsies from iSGS scar (<jats:italic>n<\/jats:italic>\u00a0=\u20094), and matched non\u2010scar (<jats:italic>n<\/jats:italic>\u00a0=\u20094) regions were analyzed using single\u2010cell RNA\u2010seq (scRNA\u2010seq). Immunofluorescence (IF) was performed on rapidly processed autopsies (RPA) and iSGS tracheal resections (<jats:italic>n<\/jats:italic>\u00a0=\u20094) to co\u2010localize <jats:italic>S100A8\/9<\/jats:italic> and <jats:italic>CD11b<\/jats:italic>. Collagen gene\/protein expression was assessed in iSGS fibroblasts (<jats:italic>n<\/jats:italic>\u00a0=\u20094) treated with protein <jats:italic>S100A8\/9<\/jats:italic> (1000\u2009ng\/ml). Macrophages were subclustered to identify distinct subpopulations.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>scRNA\u2010seq analysis revealed <jats:italic>S100A8\/S100A9<\/jats:italic> (fold change (FC)\u00a0=\u00a04.1\/1.88, <jats:italic>p<\/jats:italic>\u2009&lt;\u20090.001) as top differentially expressed genes in iSGS macrophages. IF exhibited increased <jats:italic>CD11b+\/S100A8\/9+<\/jats:italic> cells in tracheal samples of iSGS versus RPA (26.75%\u2009\u00b1\u20097.08 vs. 0.594%\u2009\u00b1\u20090.974, <jats:italic>n<\/jats:italic>\u00a0=\u00a04, <jats:italic>p<\/jats:italic>\u00a0=\u00a00.029). iSGS fibroblasts treated with <jats:italic>S100A8\/9<\/jats:italic> demonstrated increased gene expression of <jats:italic>COL1A1<\/jats:italic> (FC\u00a0=\u00a02.30\u2009\u00b1\u20090.45, <jats:italic>p<\/jats:italic>\u00a0=\u00a00.03, <jats:italic>n<\/jats:italic>\u00a0=\u00a04) and <jats:italic>COL3A1<\/jats:italic> (FC\u00a0=\u00a02.44\u2009\u00b1\u20090.40, <jats:italic>p<\/jats:italic>\u00a0=\u00a00.03, <jats:italic>n<\/jats:italic>\u00a0=\u00a04). <jats:italic>COL1A1<\/jats:italic> protein assays revealed an increase in the experimental group, albeit not significant, (<jats:italic>p<\/jats:italic>\u00a0=\u00a00.12, <jats:italic>n<\/jats:italic>\u00a0=\u00a04). Finally, macrophage sub clustering revealed one subpopulation as a predominant source of <jats:italic>S100A8\/S100A9<\/jats:italic> expression (FC\u00a0=\u00a07.94\/5.47, <jats:italic>p<\/jats:italic>\u2009&lt;\u20090.001).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p><jats:italic>S100A8\/9<\/jats:italic> is a key biomarker in iSGS macrophages. Although <jats:italic>S100A8\/9<\/jats:italic> demonstrates profibrotic nature <jats:italic>in vitro<\/jats:italic>, the role of S100A8\/9+ macrophages <jats:italic>in vivo<\/jats:italic> warrants further investigation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Level of Evidence<\/jats:title><jats:p>NA <jats:italic>Laryngoscope<\/jats:italic>, 133:2308\u20132316, 2023<\/jats:p><\/jats:sec>","DOI":"10.1002\/lary.30524","type":"journal-article","created":{"date-parts":[[2022,12,16]],"date-time":"2022-12-16T11:15:53Z","timestamp":1671189353000},"page":"2308-2316","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Characterizing the Macrophage Population in Patients With Idiopathic Subglottic Stenosis"],"prefix":"10.1002","volume":"133","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8501-1934","authenticated-orcid":false,"given":"Rafael","family":"Ospino","sequence":"first","affiliation":[{"name":"Johns Hopkins 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