{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,14]],"date-time":"2026-03-14T14:22:05Z","timestamp":1773498125165,"version":"3.50.1"},"reference-count":61,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2020,3,23]],"date-time":"2020-03-23T00:00:00Z","timestamp":1584921600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Laryngoscope"],"published-print":{"date-parts":[[2021,1]]},"abstract":"<jats:sec><jats:title>Objective<\/jats:title><jats:p>Laryngopharyngeal reflux (LPR) is a common affliction that contributes to laryngeal inflammation, symptoms that impact quality of life, and life\u2010threatening illnesses such as cancer. Effective treatment strategies for LPR are lacking. Pepsin is a proinflammatory and carcinogenic element of refluxate. Investigation of molecular pathways involved in pepsin\u2010mediated damage may lead to identification of novel biomarkers and therapeutic targets for LPR. In this study, RNA sequencing was used to examine changes in human laryngeal epithelial cells following brief pepsin insult. Cells were immortalized to generate a model to aid future study of laryngeal injury and therapeutics.<\/jats:p><\/jats:sec><jats:sec><jats:title>Study Design<\/jats:title><jats:p>In vitro translational.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Laryngeal epithelial cells were cultured from a patient without signs or symptoms of LPR or laryngeal cancer. Cells were treated with 0.1\u2009mg\/ml pepsin for 1\u2009hour or normal growth media (control) prior to RNA sequencing. Cells were immortalized via HPV E6\/7 and characterized by microscopy, immunohistochemistry, G\u2010banding, and soft agar assay.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Three hundred ninety\u2010seven genes exhibited differences in expression with pepsin treatment (<jats:italic>P<\/jats:italic> &lt;\u2009.05). Pathway analysis revealed association with cancer and related signaling processes including dysregulation of cancer\u2010associated molecules, Metastasis\u2010Associated Lung Adenocarcinoma Transcript 1 and KRT82, and the long\u2010noncoding RNA, lipoprotein receptor\u2010related protein 1 (LRP1)\u2013AS, which regulates the putative pepsin receptor LRP1.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>A single, brief exposure to pepsin activated cancer\u2010associated signaling pathways in laryngeal cells in vitro<jats:italic>,<\/jats:italic> revealing novel mechanisms by which chronic reflux may contribute to carcinogenesis. The cell line developed herein represents a novel tool in which to investigate pepsin\u2010dysregulated pathways identified by RNA sequencing and disparities of tumor proneness of laryngeal subsites.<\/jats:p><\/jats:sec><jats:sec><jats:title>Level of Evidence<\/jats:title><jats:p>N\/A <jats:italic>Laryngoscope<\/jats:italic>, 131:121\u2013129, 2021<\/jats:p><\/jats:sec>","DOI":"10.1002\/lary.28636","type":"journal-article","created":{"date-parts":[[2020,3,23]],"date-time":"2020-03-23T14:14:02Z","timestamp":1584972842000},"page":"121-129","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["RNA Sequencing Reveals Cancer\u2010Associated Changes in Laryngeal Cells Exposed to Non\u2010Acid Pepsin"],"prefix":"10.1002","volume":"131","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7045-357X","authenticated-orcid":false,"given":"Tina L.","family":"Samuels","sequence":"first","affiliation":[{"name":"Department of Otolaryngology and Communication Sciences Medical College of Wisconsin  Milwaukee Wisconsin U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael T.","family":"Zimmermann","sequence":"additional","affiliation":[{"name":"Bioinformatics Research and Development Laboratory Genomic Sciences and Precision Medicine Center, Medical College of Wisconsin  Milwaukee Wisconsin U.S.A."},{"name":"Clinical and Translational Science Institute Medical College of Wisconsin  Milwaukee Wisconsin U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Atefeh","family":"Zeighami","sequence":"additional","affiliation":[{"name":"Bioinformatics Research and Development Laboratory Genomic Sciences and Precision Medicine Center, Medical College of Wisconsin  Milwaukee Wisconsin U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wendy","family":"Demos","sequence":"additional","affiliation":[{"name":"Bioinformatics Research and Development Laboratory Genomic Sciences and Precision Medicine Center, Medical College of Wisconsin  Milwaukee Wisconsin U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jessica E.","family":"Southwood","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology and Communication Sciences Medical College of Wisconsin  Milwaukee Wisconsin U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joel H.","family":"Blumin","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology and Communication Sciences Medical College of Wisconsin  Milwaukee Wisconsin U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4367-7527","authenticated-orcid":false,"given":"Jonathan M.","family":"Bock","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology and Communication Sciences Medical College of Wisconsin  Milwaukee Wisconsin U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nikki","family":"Johnston","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology and Communication Sciences Medical College of Wisconsin  Milwaukee Wisconsin U.S.A."},{"name":"Department of Microbiology and Immunology Medical College of Wisconsin  Milwaukee Wisconsin U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,3,23]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1001\/jamaoto.2017.2561"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.3342\/ceo.2011.4.2.101"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/lary.1991.101.s53.1"},{"key":"e_1_2_6_5_1","first-page":"10","article-title":"Symptoms and findings of laryngopharyngeal reflux","volume":"81","author":"Belafsky PC","year":"2002","journal-title":"Ear Nose Throat 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