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

Research

Spadaro Francesca¹  Cimino Giuseppe²  Blaconà Giovanna³  Bruno Sabina Maria³  Lucarelli Marco⁴  Esposito Speranza⁵  Villella Valeria Rachela⁵  Amato Felice⁵  Zollo Immacolata⁵  Eramo Adriana⁶  Castelli Germana⁶  Lo Cicero Stefania⁶  Pigliucci Riccardo⁶  Sette Giovanni⁶  Biffoni Mauro⁶  De Maria Ruggero⁷ 

[1] Confocal Microscopy Unit, Core Facilities, Istituto Superiore di Sanità, Rome, Italy;Cystic Fibrosis Reference Center of Lazio Region, AOU Policlinico Umberto I, Rome, Italy;Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy;Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy;Pasteur Institute, Cenci Bolognetti Foundation, Sapienza University of Rome, Rome, Italy;Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, Naples, Italy;CEINGE-Biotecnologie Avanzate S.c.a.r.l, Naples, Italy;Department of Oncology and Molecular Medicine, Istituto Superiore di Sanità, Rome, Italy;Dipartimento di Medicina e Chirurgia Traslazionale, Università Cattolica del Sacro Cuore, Rome, Italy;Fondazione Policlinico Universitario ‘A. Gemelli’-IRCCS, Rome, Italy;

关键词: Cystic fibrosis;    CFTR;    Reprogrammed nasal cells;    Nasal organoids;    ALI culture;    Trikafta;    Rare mutation;    Theratyping;    Personalized medicine;    Patient-derived models;   

DOI  :  10.1186/s12931-023-02516-0

 received in 2022-11-14, accepted in 2023-08-17,  发布年份 2023

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY   
© BioMed Central Ltd., part of Springer Nature 2023

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