L1077P CFTR pathogenic variant function rescue by Elexacaftor–Tezacaftor–Ivacaftor in cystic fibrosis patient-derived air–liquid interface (ALI) cultures and organoids: in vitro guided personalized therapy of non-F508del patients
Abstract Cystic fibrosis (CF) is caused by defects of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR-modulating drugs may overcome specific defects, such as the case of Trikafta, which is a clinically approved triple combination of Elexacaftor, Tezacaftor and Ivacaftor (ET...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-09-01
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| Series: | Respiratory Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12931-023-02516-0 |
| _version_ | 1827708836602970112 |
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| author | Stefania Lo Cicero Germana Castelli Giovanna Blaconà Sabina Maria Bruno Giovanni Sette Riccardo Pigliucci Valeria Rachela Villella Speranza Esposito Immacolata Zollo Francesca Spadaro Ruggero De Maria Mauro Biffoni Giuseppe Cimino Felice Amato Marco Lucarelli Adriana Eramo |
| author_facet | Stefania Lo Cicero Germana Castelli Giovanna Blaconà Sabina Maria Bruno Giovanni Sette Riccardo Pigliucci Valeria Rachela Villella Speranza Esposito Immacolata Zollo Francesca Spadaro Ruggero De Maria Mauro Biffoni Giuseppe Cimino Felice Amato Marco Lucarelli Adriana Eramo |
| author_sort | Stefania Lo Cicero |
| collection | DOAJ |
| description | Abstract Cystic fibrosis (CF) is caused by defects of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR-modulating drugs may overcome specific defects, such as the case of Trikafta, which is a clinically approved triple combination of Elexacaftor, Tezacaftor and Ivacaftor (ETI) that exhibited a strong ability to rescue the function of the most frequent F508del pathogenic variant even in genotypes with the mutated allele in single copy. Nevertheless, most rare genotypes lacking the F508del allele are still not eligible for targeted therapies. Via the innovative approach of using nasal conditionally reprogrammed cell (CRC) cell-based models that mimic patient disease in vitro, which are obtainable from each patient due to the 100% efficiency of the cell culture establishment, we theratyped orphan CFTR mutation L1077P. Protein studies, Forskolin-induced organoid swelling, and Ussing chamber assays congruently proved the L1077P variant function rescue by ETI. Notably, this rescue takes place even in the context of a single-copy L1077P allele, which appears to enhance its expression. Thus, the possibility of single-allele treatment also arises for rare genotypes, with an allele-specific modulation as part of the mechanism. Of note, besides providing indication of drug efficacy with respect to specific CFTR pathogenic variants or genotypes, this approach allows the evaluation of the response of single-patient cells within their genetic background. In this view, our studies support in vitro guided personalized CF therapies also for rare patients who are nearly excluded from clinical trials. |
| first_indexed | 2024-03-10T17:10:27Z |
| format | Article |
| id | doaj.art-b4d7cae7df80409786f2da0722aa6ceb |
| institution | Directory Open Access Journal |
| issn | 1465-993X |
| language | English |
| last_indexed | 2024-03-10T17:10:27Z |
| publishDate | 2023-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Respiratory Research |
| spelling | doaj.art-b4d7cae7df80409786f2da0722aa6ceb2023-11-20T10:40:48ZengBMCRespiratory Research1465-993X2023-09-0124112010.1186/s12931-023-02516-0L1077P CFTR pathogenic variant function rescue by Elexacaftor–Tezacaftor–Ivacaftor in cystic fibrosis patient-derived air–liquid interface (ALI) cultures and organoids: in vitro guided personalized therapy of non-F508del patientsStefania Lo Cicero0Germana Castelli1Giovanna Blaconà2Sabina Maria Bruno3Giovanni Sette4Riccardo Pigliucci5Valeria Rachela Villella6Speranza Esposito7Immacolata Zollo8Francesca Spadaro9Ruggero De Maria10Mauro Biffoni11Giuseppe Cimino12Felice Amato13Marco Lucarelli14Adriana Eramo15Department of Oncology and Molecular Medicine, Istituto Superiore di SanitàDepartment of Oncology and Molecular Medicine, Istituto Superiore di SanitàDepartment of Experimental Medicine, Sapienza University of RomeDepartment of Experimental Medicine, Sapienza University of RomeDepartment of Oncology and Molecular Medicine, Istituto Superiore di SanitàDepartment of Oncology and Molecular Medicine, Istituto Superiore di SanitàDepartment of Molecular Medicine and Medical Biotechnologies, University of Naples Federico IIDepartment of Molecular Medicine and Medical Biotechnologies, University of Naples Federico IIDepartment of Molecular Medicine and Medical Biotechnologies, University of Naples Federico IIConfocal Microscopy Unit, Core Facilities, Istituto Superiore di SanitàDipartimento di Medicina e Chirurgia Traslazionale, Università Cattolica del Sacro CuoreDepartment of Oncology and Molecular Medicine, Istituto Superiore di SanitàCystic Fibrosis Reference Center of Lazio Region, AOU Policlinico Umberto IDepartment of Molecular Medicine and Medical Biotechnologies, University of Naples Federico IIDepartment of Experimental Medicine, Sapienza University of RomeDepartment of Oncology and Molecular Medicine, Istituto Superiore di SanitàAbstract Cystic fibrosis (CF) is caused by defects of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR-modulating drugs may overcome specific defects, such as the case of Trikafta, which is a clinically approved triple combination of Elexacaftor, Tezacaftor and Ivacaftor (ETI) that exhibited a strong ability to rescue the function of the most frequent F508del pathogenic variant even in genotypes with the mutated allele in single copy. Nevertheless, most rare genotypes lacking the F508del allele are still not eligible for targeted therapies. Via the innovative approach of using nasal conditionally reprogrammed cell (CRC) cell-based models that mimic patient disease in vitro, which are obtainable from each patient due to the 100% efficiency of the cell culture establishment, we theratyped orphan CFTR mutation L1077P. Protein studies, Forskolin-induced organoid swelling, and Ussing chamber assays congruently proved the L1077P variant function rescue by ETI. Notably, this rescue takes place even in the context of a single-copy L1077P allele, which appears to enhance its expression. Thus, the possibility of single-allele treatment also arises for rare genotypes, with an allele-specific modulation as part of the mechanism. Of note, besides providing indication of drug efficacy with respect to specific CFTR pathogenic variants or genotypes, this approach allows the evaluation of the response of single-patient cells within their genetic background. In this view, our studies support in vitro guided personalized CF therapies also for rare patients who are nearly excluded from clinical trials.https://doi.org/10.1186/s12931-023-02516-0Cystic fibrosisCFTRReprogrammed nasal cellsNasal organoidsALI cultureTrikafta |
| spellingShingle | Stefania Lo Cicero Germana Castelli Giovanna Blaconà Sabina Maria Bruno Giovanni Sette Riccardo Pigliucci Valeria Rachela Villella Speranza Esposito Immacolata Zollo Francesca Spadaro Ruggero De Maria Mauro Biffoni Giuseppe Cimino Felice Amato Marco Lucarelli Adriana Eramo L1077P CFTR pathogenic variant function rescue by Elexacaftor–Tezacaftor–Ivacaftor in cystic fibrosis patient-derived air–liquid interface (ALI) cultures and organoids: in vitro guided personalized therapy of non-F508del patients Respiratory Research Cystic fibrosis CFTR Reprogrammed nasal cells Nasal organoids ALI culture Trikafta |
| title | L1077P CFTR pathogenic variant function rescue by Elexacaftor–Tezacaftor–Ivacaftor in cystic fibrosis patient-derived air–liquid interface (ALI) cultures and organoids: in vitro guided personalized therapy of non-F508del patients |
| title_full | L1077P CFTR pathogenic variant function rescue by Elexacaftor–Tezacaftor–Ivacaftor in cystic fibrosis patient-derived air–liquid interface (ALI) cultures and organoids: in vitro guided personalized therapy of non-F508del patients |
| title_fullStr | L1077P CFTR pathogenic variant function rescue by Elexacaftor–Tezacaftor–Ivacaftor in cystic fibrosis patient-derived air–liquid interface (ALI) cultures and organoids: in vitro guided personalized therapy of non-F508del patients |
| title_full_unstemmed | L1077P CFTR pathogenic variant function rescue by Elexacaftor–Tezacaftor–Ivacaftor in cystic fibrosis patient-derived air–liquid interface (ALI) cultures and organoids: in vitro guided personalized therapy of non-F508del patients |
| title_short | L1077P CFTR pathogenic variant function rescue by Elexacaftor–Tezacaftor–Ivacaftor in cystic fibrosis patient-derived air–liquid interface (ALI) cultures and organoids: in vitro guided personalized therapy of non-F508del patients |
| title_sort | l1077p cftr pathogenic variant function rescue by elexacaftor tezacaftor ivacaftor in cystic fibrosis patient derived air liquid interface ali cultures and organoids in vitro guided personalized therapy of non f508del patients |
| topic | Cystic fibrosis CFTR Reprogrammed nasal cells Nasal organoids ALI culture Trikafta |
| url | https://doi.org/10.1186/s12931-023-02516-0 |
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