{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,7]],"date-time":"2026-05-07T22:19:07Z","timestamp":1778192347143,"version":"3.51.4"},"reference-count":25,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2025,3,6]],"date-time":"2025-03-06T00:00:00Z","timestamp":1741219200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["physoc.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Physiology"],"published-print":{"date-parts":[[2025,5]]},"abstract":"<jats:sec><jats:title>Abstract<\/jats:title><jats:p>Electrogenic transepithelial ion transport can be measured with the short\u2010circuit current technique. Such experiments are frequently used to evaluate the activity of the cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP\u2010activated chloride channel that is defective in cystic fibrosis, one of the most frequent genetic diseases. Typically, CFTR activity is estimated from the effect of CFTR<jats:sub>inh<\/jats:sub>\u2010172, a selective CFTR inhibitor. Unexpectedly, we found that CFTR<jats:sub>inh<\/jats:sub>\u2010172, in addition to PPQ\u2010102, another CFTR inhibitor, caused only partial inhibition of CFTR function, particularly in epithelia in pro\u2010inflammatory conditions, which are characterized by abundant mucus secretion. We hypothesized that the mucus layer was responsible for the poor activity of CFTR inhibitors. Therefore, we treated the epithelial surface with the reducing agent dithiothreitol to remove mucus. Removal of mucus, confirmed by immunofluorescence, resulted in highly enhanced sensitivity of CFTR to pharmacological inhibition. Our results show that the mucus layer represents an important barrier whose presence limits the activity of pharmacological agents. This is particularly relevant for CFTR and the evaluation of therapeutic approaches for correction of the basic defect in cystic fibrosis.\n<jats:boxed-text content-type=\"graphic\" position=\"anchor\"><jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" mimetype=\"image\/png\" position=\"anchor\" specific-use=\"enlarged-web-image\" xlink:href=\"graphic\/tjp16615-gra-0001-m.png\"><jats:alt-text>image<\/jats:alt-text><\/jats:graphic><\/jats:boxed-text><\/jats:p><\/jats:sec><jats:sec><jats:title>Key points<\/jats:title><jats:p><jats:list list-type=\"bullet\">\n<jats:list-item><jats:p>Activity of the cAMP\u2010activated cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel can be evaluated by measuring the inhibition elicited by the selective blockers CFTR<jats:sub>inh<\/jats:sub>\u2010172 and PPQ\u2010102.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>In short\u2010circuit current recordings on human airway epithelia, CFTR inhibitors had only a partial effect on cAMP\u2010dependent chloride secretion, suggesting the possible contribution of other ion channels.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>The mucus layer covering the epithelial surface was removed with the reducing agent dithiothreitol.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>Treatment of epithelia with dithiothreitol markedly improved the efficacy of CFTR inhibitors.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>The partial effect of CFTR inhibitors might be explained by the presence of the mucus layer acting as a barrier.<\/jats:p><\/jats:list-item>\n<\/jats:list><\/jats:p><\/jats:sec>","DOI":"10.1113\/jp287891","type":"journal-article","created":{"date-parts":[[2025,3,12]],"date-time":"2025-03-12T04:19:47Z","timestamp":1741753187000},"page":"2619-2632","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["The apical mucus layer alters the pharmacological properties of the airway epithelium"],"prefix":"10.1113","volume":"603","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9301-8095","authenticated-orcid":false,"given":"Daniela","family":"Guidone","sequence":"first","affiliation":[{"name":"Telethon Institute of Genetics and Medicine  Pozzuoli Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-8185-6490","authenticated-orcid":false,"given":"Martina","family":"de Santis","sequence":"additional","affiliation":[{"name":"Telethon Institute of Genetics and Medicine  Pozzuoli Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emanuela","family":"Pesce","sequence":"additional","affiliation":[{"name":"UOC Genetica Medica Istituto Giannina Gaslini  Genova Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valeria","family":"Capurro","sequence":"additional","affiliation":[{"name":"UOC Genetica Medica Istituto Giannina Gaslini  Genova Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5161-1720","authenticated-orcid":false,"given":"Nicoletta","family":"Pedemonte","sequence":"additional","affiliation":[{"name":"UOC Genetica Medica Istituto Giannina Gaslini  Genova Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8686-3461","authenticated-orcid":false,"given":"Luis J. V.","family":"Galietta","sequence":"additional","affiliation":[{"name":"Telethon Institute of Genetics and Medicine  Pozzuoli Italy"},{"name":"Department of Translational Medical Sciences University of Napoli \u2018Federico II\u2019  Naples Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,3,6]]},"reference":[{"key":"e_1_2_5_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2023.03.004"},{"key":"e_1_2_5_3_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.200810097"},{"key":"e_1_2_5_4_1","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20080029"},{"key":"e_1_2_5_5_1","doi-asserted-by":"publisher","DOI":"10.1183\/13993003.00110-2023"},{"key":"e_1_2_5_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(16)00576-6"},{"key":"e_1_2_5_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep36016"},{"issue":"22","key":"e_1_2_5_8_1","article-title":"Airway surface hyperviscosity and defective mucociliary transport by IL\u201017\/TNF\u2010\u03b1 are corrected by \u03b2\u2010adrenergic stimulus","volume":"7","author":"Guidone D.","year":"2022","journal-title":"Journal of Clinical Investigation Insight"},{"key":"e_1_2_5_9_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1807120"},{"key":"e_1_2_5_10_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI0215217"},{"key":"e_1_2_5_11_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajplung.00563.2020"},{"key":"e_1_2_5_12_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2019.01662"},{"key":"e_1_2_5_13_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI0216112"},{"key":"e_1_2_5_14_1","doi-asserted-by":"publisher","DOI":"10.1089\/jamp.2007.0659"},{"key":"e_1_2_5_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2016.05.012"},{"key":"e_1_2_5_16_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aay9669"},{"key":"e_1_2_5_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-023-34440-0"},{"key":"e_1_2_5_18_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI181995"},{"key":"e_1_2_5_19_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells9092137"},{"key":"e_1_2_5_20_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.2012.240838"},{"key":"e_1_2_5_21_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells9092090"},{"key":"e_1_2_5_22_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms23063175"},{"key":"e_1_2_5_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcf.2022.12.005"},{"key":"e_1_2_5_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiomech.2019.109578"},{"key":"e_1_2_5_25_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0904709106"},{"key":"e_1_2_5_26_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2023.1293578"}],"container-title":["The Journal of Physiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/physoc.onlinelibrary.wiley.com\/doi\/pdf\/10.1113\/JP287891","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,5,27]],"date-time":"2025-05-27T05:35:33Z","timestamp":1748324133000},"score":1,"resource":{"primary":{"URL":"https:\/\/physoc.onlinelibrary.wiley.com\/doi\/10.1113\/JP287891"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,6]]},"references-count":25,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2025,5]]}},"alternative-id":["10.1113\/JP287891"],"URL":"https:\/\/doi.org\/10.1113\/jp287891","archive":["Portico"],"relation":{},"ISSN":["0022-3751","1469-7793"],"issn-type":[{"value":"0022-3751","type":"print"},{"value":"1469-7793","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,6]]},"assertion":[{"value":"2024-10-16","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-02-11","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-03-06","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}