{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T08:10:00Z","timestamp":1774512600493,"version":"3.50.1"},"reference-count":34,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,3,8]],"date-time":"2023-03-08T00:00:00Z","timestamp":1678233600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008385","name":"Italian Cystic Fibrosis Research Foundation (FFC)","doi-asserted-by":"publisher","award":["FFC #13\/2018"],"award-info":[{"award-number":["FFC #13\/2018"]}],"id":[{"id":"10.13039\/501100008385","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008385","name":"Italian Cystic Fibrosis Research Foundation (FFC)","doi-asserted-by":"publisher","award":["FFC #9\/2020"],"award-info":[{"award-number":["FFC #9\/2020"]}],"id":[{"id":"10.13039\/501100008385","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008385","name":"Italian Cystic Fibrosis Research Foundation (FFC)","doi-asserted-by":"publisher","award":["Assael08A0"],"award-info":[{"award-number":["Assael08A0"]}],"id":[{"id":"10.13039\/501100008385","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Lega Italiana Fibrosi Cistica onlus","award":["FFC #13\/2018"],"award-info":[{"award-number":["FFC #13\/2018"]}]},{"name":"Lega Italiana Fibrosi Cistica onlus","award":["FFC #9\/2020"],"award-info":[{"award-number":["FFC #9\/2020"]}]},{"name":"Lega Italiana Fibrosi Cistica onlus","award":["Assael08A0"],"award-info":[{"award-number":["Assael08A0"]}]},{"DOI":"10.13039\/100006501","name":"American CFF","doi-asserted-by":"publisher","award":["FFC #13\/2018"],"award-info":[{"award-number":["FFC #13\/2018"]}],"id":[{"id":"10.13039\/100006501","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006501","name":"American CFF","doi-asserted-by":"publisher","award":["FFC #9\/2020"],"award-info":[{"award-number":["FFC #9\/2020"]}],"id":[{"id":"10.13039\/100006501","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006501","name":"American CFF","doi-asserted-by":"publisher","award":["Assael08A0"],"award-info":[{"award-number":["Assael08A0"]}],"id":[{"id":"10.13039\/100006501","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>An Italian, 46-year-old female patient carrying the complex allele p.[R74W;V201M;D1270N] in trans with CFTR dele22_24 was diagnosed at the Cystic Fibrosis (CF) Center of Verona as being affected by CF-pancreatic sufficient (CF-PS) in 2021. The variant V201M has unknown significance, while both of the other variants of this complex allele have variable clinical consequences, according to the CFTR2 database, with reported clinical benefits for treatment with ivacaftor + tezacaftor and ivacaftor + tezacaftor + elexacaftor in patients carrying the R74W-D1270N complex allele, which are currently approved (in USA, not yet in Italy). She was previously followed up by pneumologists in northern Italy because of frequent bronchitis, hemoptysis, recurrent rhinitis, Pseudomonas aeruginosa lung colonization, bronchiectasis\/atelectasis, bronchial arterial embolization and moderately compromised lung function (FEV1: 62%). Following a sweat test with borderline results, she was referred to the Verona CF Center where she presented abnormal values in both optical beta-adrenergic sweat tests and intestinal current measurement (ICM). These results were consistent with a diagnosis of CF. CFTR function analyses were also performed in vitro by forskolin-induced swelling (FIS) assay and short-circuit currents (Isc) in the monolayers of the rectal organoids. Both of these assays showed significantly increased CFTR activity following treatment with the CFTR modulators. Western-blot analysis revealed increased fully glycosylated CFTR protein after treatment with correctors, in line with the functional analysis. Interestingly, tezacaftor, together with elexacaftor, rescued the total organoid area under steady-state conditions, even in the absence of the CFTR agonist forskolin. In conclusion, in ex vivo and in vitro assays, we measured a residual function that was significantly enhanced by in vitro incubation with CFTR modulators, especially by ivacaftor + tezacaftor + elexacaftor, suggesting this combination as a potentially optimal treatment for this case.<\/jats:p>","DOI":"10.3390\/ijms24065199","type":"journal-article","created":{"date-parts":[[2023,3,9]],"date-time":"2023-03-09T02:33:16Z","timestamp":1678329196000},"page":"5199","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["CFTR Modulators Rescue the Activity of CFTR in Colonoids Expressing the Complex Allele p.[R74W;V201M;D1270N]\/dele22_24"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1267-4814","authenticated-orcid":false,"given":"Karina","family":"Kleinfelder","sequence":"first","affiliation":[{"name":"Department of Medicine, Division of General Pathology, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elena","family":"Somenza","sequence":"additional","affiliation":[{"name":"Department of Medicine, Division of General Pathology, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessia","family":"Farinazzo","sequence":"additional","affiliation":[{"name":"Department of Medicine, Division of General Pathology, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4996-4463","authenticated-orcid":false,"given":"Jessica","family":"Conti","sequence":"additional","affiliation":[{"name":"Department of Medicine, Division of General Pathology, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5746-3810","authenticated-orcid":false,"given":"Virginia","family":"Lotti","sequence":"additional","affiliation":[{"name":"Department of Medicine, Division of General Pathology, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6722-5683","authenticated-orcid":false,"given":"Roberta Valeria","family":"Latorre","sequence":"additional","affiliation":[{"name":"Department of Medicine, Division of General Pathology, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luca","family":"Rodella","sequence":"additional","affiliation":[{"name":"Endoscopic Surgery Unit, Azienda Ospedaliera Universitaria Integrata Verona, 37126 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arianna","family":"Massella","sequence":"additional","affiliation":[{"name":"Endoscopic Surgery Unit, Azienda Ospedaliera Universitaria Integrata Verona, 37126 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesco","family":"Tomba","sequence":"additional","affiliation":[{"name":"Endoscopic Surgery Unit, Azienda Ospedaliera Universitaria Integrata Verona, 37126 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marina","family":"Bertini","sequence":"additional","affiliation":[{"name":"Cystic Fibrosis Centre, Azienda Ospedaliera Universitaria Integrata Verona, Piazzale Stefani, 1, 37126 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2739-4014","authenticated-orcid":false,"given":"Claudio","family":"Sorio","sequence":"additional","affiliation":[{"name":"Department of Medicine, Division of General Pathology, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5276-1595","authenticated-orcid":false,"given":"Paola","family":"Melotti","sequence":"additional","affiliation":[{"name":"Cystic Fibrosis Centre, Azienda Ospedaliera Universitaria Integrata Verona, Piazzale Stefani, 1, 37126 Verona, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Scotet, V., Gutierrez, H., and Farrell, P.M. (2020). Newborn screening for CF across the globe\u2014Where is it worthwhile?. Int. J. Neonatal Screen., 6.","DOI":"10.3390\/ijns6010018"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1038\/s41390-019-0503-8","article-title":"Image-based \u03b2-adrenergic sweat rate assay captures minimal cystic fibrosis transmembrane conductance regulator function","volume":"87","author":"Salinas","year":"2020","journal-title":"Pediatr. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1097\/GIM.0b013e3181ead634","article-title":"A new complex allele of the CFTR gene partially explains the variable phenotype of the L997F mutation","volume":"12","author":"Lucarelli","year":"2010","journal-title":"Genet. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"S15","DOI":"10.1016\/j.jcf.2019.12.008","article-title":"The influence of CFTR complex alleles on precision therapy of cystic fibrosis","volume":"19","author":"Chevalier","year":"2020","journal-title":"J. Cyst. Fibros."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1002\/humu.23389","article-title":"Cis variants identified in F508del complex alleles modulate CFTR channel rescue by small molecules","volume":"39","author":"Baatallah","year":"2018","journal-title":"Hum. Mutat."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Ramalho, S.S., Silva, I.A.L., Amaral, M.D., and Farinha, C.M. (2022). Rare Trafficking CFTR Mutations Involve Distinct Cellular Retention Machineries and Require Different Rescuing Strategies. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23010024"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Scudieri, P., Musante, I., Venturini, A., Guidone, D., Genovese, M., Cresta, F., Caci, E., Palleschi, A., Poeta, M., and Santamaria, F. (2020). Ionocytes and CFTR Chloride Channel Expression in Normal and Cystic Fibrosis Nasal and Bronchial Epithelial Cells. Cells, 9.","DOI":"10.3390\/cells9092090"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"S60","DOI":"10.1016\/j.jcf.2019.11.002","article-title":"Intestinal organoids for Cystic Fibrosis research","volume":"19","author":"Lefferts","year":"2020","journal-title":"J. Cyst. Fibros."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.jcf.2018.05.004","article-title":"CFTR modulator theratyping: Current status, gaps and future directions","volume":"18","author":"Clancy","year":"2019","journal-title":"J. Cyst. Fibros."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Silva, I.A.L., Laselva, O., and Lopes-Pacheco, M. (2022). Advances in Preclinical In Vitro Models for the Translation of Precision Medicine for Cystic Fibrosis. J. Pers. Med., 12.","DOI":"10.3390\/jpm12081321"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1016\/j.jcf.2010.08.014","article-title":"[R74W;R1070W;D1270N]: A new complex allele responsible for cystic fibrosis","volume":"9","author":"Merino","year":"2010","journal-title":"J. Cyst. Fibros."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2004.e23","DOI":"10.1016\/j.fertnstert.2008.05.057","article-title":"Outcome of intracytoplasmic sperm injection for a couple in which the man is carrier of CFTR p.[R74W;V201M;D1270N] and p.P841R mutations and his spouse a heterozygous carrier of p.F508del mutation of the cystic fibrosis transmembrane conductance regulator gene","volume":"90","author":"Brugnon","year":"2008","journal-title":"Fertil. Steril."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Taulan, M., Girardet, A., Guittard, C., Altieri, J.-P., Templin, C., Beroud, C., Georges, M.D., and Claustres, M. (2007). Large genomic rearrangements in the CFTRgene contribute to CBAVD. BMC Med. Genet., 8.","DOI":"10.1186\/1471-2350-8-22"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e000736","DOI":"10.1136\/bmjresp-2020-000736","article-title":"Intestinal current measurement and nasal potential difference to make a diagnosis of cases with inconclusive CFTR genetics and sweat test","volume":"7","author":"Minso","year":"2020","journal-title":"BMJ Open Respir. Res."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Wine, J.J., Char, J.E., Chen, J., Cho, H.-J., Dunn, C., Frisbee, E., Joo, N.S., Milla, C., Modlin, S.E., and Park, I.-H. (2013). In Vivo Readout of CFTR Function: Ratiometric Measurement of CFTR-Dependent Secretion by Individual, Identifiable Human Sweat Glands. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0077114"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1016\/j.jcf.2017.12.003","article-title":"Ratiometric sweat secretion optical test in cystic fibrosis, carriers and healthy subjects","volume":"17","author":"Bergamini","year":"2018","journal-title":"J. Cyst. Fibros."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1763","DOI":"10.1172\/JCI111385","article-title":"Defective beta adrenergic response of cystic fibrosis sweat glands in vivo and in vitro","volume":"73","author":"Sato","year":"1984","journal-title":"J. Clin. Investig."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1203\/00006450-198709000-00007","article-title":"Hyposecretion of beta-adrenergically induced sweating in cystic fibrosis heterozygotes","volume":"22","author":"Behm","year":"1987","journal-title":"Pediatr. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1038\/nm.3201","article-title":"A functional CFTR assay using primary cystic fibrosis intestinal organoids","volume":"19","author":"Dekkers","year":"2013","journal-title":"Nat. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"344ra84","DOI":"10.1126\/scitranslmed.aad8278","article-title":"Characterizing responses to CFTR-modulating drugs using rectal organoids derived from subjects with cystic fibrosis","volume":"8","author":"Dekkers","year":"2016","journal-title":"Sci. Transl. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1701","DOI":"10.1016\/j.celrep.2019.01.068","article-title":"Rectal organoids enable personalized treatment of cystic fibrosis","volume":"26","author":"Berkers","year":"2019","journal-title":"Cell Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1802379","DOI":"10.1183\/13993003.02379-2018","article-title":"Intestinal organoids to model cystic fibrosis","volume":"54","year":"2019","journal-title":"Eur. Respir. J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"594","DOI":"10.1136\/thx.2009.125088","article-title":"Intestinal current measurement for diagnostic classification of patients with questionable cystic fibrosis: Validation and reference data","volume":"65","author":"Derichs","year":"2010","journal-title":"Thorax"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Rueden, C.T., Schindelin, J., Hiner, M.C., DeZonia, B.E., Walter, A.E., Arena, E.T., and Eliceiri, K.W. (2017). ImageJ2: ImageJ for the next generation of scientific image data. BMC Bioinform., 18.","DOI":"10.1186\/s12859-017-1934-z"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1016\/S0014-5793(99)00647-X","article-title":"Structure-function analysis of a double-mutant cystic fibrosis transmembrane conductance regulator protein occurring in disorders related to cystic fibrosis","volume":"452","author":"Fanen","year":"1999","journal-title":"FEBS Lett."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"873","DOI":"10.1002\/humu.22300","article-title":"Combined computational-experimental analyses of CFTR exon strength uncover predictability of exon-skipping level","volume":"34","author":"Aissat","year":"2013","journal-title":"Hum. Mutat."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Claustres, M., Alti\u00e8ri, J.P., Guittard, C., Templin, C., Chevalier-Porst, F., and Des Georges, M. (2004). Are p.I148T, p.R74W and p.D1270N cystic fibrosis causing mutations?. BMC Med. Genet., 2.","DOI":"10.1186\/1471-2350-5-19"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Sondo, E., Cresta, F., Pastorino, C., Tomati, V., Capurro, V., Pesce, E., Lena, M., Iacomino, M., Baffico, A.M., and Coviello, D. (2022). The L467F-F508del Complex Allele Hampers Pharmacological Rescue of Mutant CFTR by Elexacaftor\/Tezacaftor\/Ivacaftor in Cystic Fibrosis Patients: The Value of the Ex Vivo Nasal Epithelial Model to Address Non-Responders to CFTR-Modulating Drugs. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23063175"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Amico, G., Brandas, C., Moran, O., and Baroni, D. (2019). Unravelling the Regions of Mutant F508del-CFTR More Susceptible to the Action of Four Cystic Fibrosis Correctors. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20215463"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1612","DOI":"10.1056\/NEJMoa1807120","article-title":"VX-445-Tezacaftor-Ivacaftor in Patients with Cystic Fibrosis and One or Two Phe508del Alleles","volume":"379","author":"Keating","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2002774","DOI":"10.1183\/13993003.02774-2020","article-title":"Rescue of multiple class II CFTR mutations by elexacaftor+tezacaftor+ivacaftor mediated in part by the dual activities of elexacaftor as both corrector and potentiator","volume":"57","author":"Laselva","year":"2021","journal-title":"Eur. Respir. J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1016\/j.jcf.2021.03.011","article-title":"Elexacaftor co-potentiates the activity of F508del and gating mutants of CFTR","volume":"20","author":"Veit","year":"2021","journal-title":"J. Cyst. Fibros."},{"key":"ref_33","first-page":"1","article-title":"Rescue by elexacaftor-tezacaftor-ivacaftor of the G1244E cystic fibrosis mutation\u2019s stability and gating defects are dependent on cell background","volume":"13","author":"Tomati","year":"2022","journal-title":"J. Cyst. Fibros."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"368","DOI":"10.1016\/j.jcf.2018.12.011","article-title":"Predictive factors for lumacaftor\/ivacaftor clinical response","volume":"18","author":"Masson","year":"2019","journal-title":"J. Cyst. Fibros."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/6\/5199\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:51:08Z","timestamp":1760122268000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/6\/5199"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,8]]},"references-count":34,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["ijms24065199"],"URL":"https:\/\/doi.org\/10.3390\/ijms24065199","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,8]]}}}