Risultato Progetto: FFC#13/2018
Analisi di organoidi intestinali per la predizione della risposta a potenziatori e correttori di CFTR utilizzati in clinica

I ricercatori riportano la creazione di organoidi come modelli sperimentali per predire la risposta a potenziatori e correttori di CFTR utilizzati in ambito clinico. Sono stati sviluppati organoidi rettali umani dalle cripte intestinali (colonoidi) da 47 pazienti FC portatori di differenti varianti del gene CFTR, appartenenti a tutte le classi di mutazioni. Sono stati condotti diversi test per valutare la funzionalità della proteina CFTR in questi modelli e per valutare la sua espressione e localizzazione nella membrana cellulare. La funzionalità di CFTR in questi colonoidi è stata anche valutata in risposta alla somministrazione di farmaci (VX-809, VX-661, VX-445 e VX-709) e composti sperimentali, attraverso l’uso del test da rigonfiamento (Forskolin-induced swelling o saggio FIS) e con la misurazione del trasporto di ioni. Questi modelli cellulari potrebbero essere utilizzati per personalizzare i trattamenti farmacologici in funzione del genotipo, anche raro del paziente (terapia personalizzata o teratipizzazione).

Researchers report the development of organoids as experimental models to predict the response to CFTR modulators used in the clinical setting. Organoids (colonoids) were developed from rectal cells from 47 CF patients carrying different variants of the CFTR gene. Several tests were conducted to evaluate the functionality of CFTR in these models and to evaluate the expression of the CFTR protein and its subcellular localization. The functionality of CFTR in these colonoids was also evaluated in response to the administration of drugs (VX-809, VX-661, VX-445 and VX-7709) and experimental compounds, through the use of the swelling test (FIS) and with the measurement of ion transport (Isc). These cellular models could be used to customize drug therapy (personalized therapy or theratyping) according to the patient’s genotype. In addition, these models could also be effective for patients carrying rare variants.

Pubblicazioni

• Caldrer S, Bergamini G, Sandri A et al. Cystic fibrosis transmembrane conductance regulator functional evaluations in a G542X+/- IVS8Tn:T7/9 patient with acute recurrent pancreatitis, World Journal of Clinical Cases, 2019 Nov 26;7(22):3757-3764

Congress abstracts

• Conti J, Kleinfelder K, Lotti V et al. Functional characterization of c.1584+18672bpA>G/2183AA>G CFTR variant in rectal organoids, 43rd European Cystic Fibrosis Conference – 2020 (virtual conference)
• Kleinfelder K, Bertini M, Vicentini R et al. Preliminary Evaluation of Corrector/Potentiator of CFTR in Patients with Cystic Fibrosis (CF) Homozygous for F508del mutation by Ratiometric Sweat Secretion Optical (RSSO) Test, 16th Annual Meeting European Cystic Fibrosis Society Diagnostic Network Working Group, 14-16 February 2019 – Tunis, Tunisia
• Kleinfelder K, Lecca P, Bertini M et al. Use of Optical Ratiometric Rate Sweat test for evaluating CFTR function in cystic fibrosis patients, 12th European CF Young Investigators Meeting (EYIM), Paris, 27 February – 1 March 2019
• Lecca P, Bertini M, Vicentini R et al. Multilinear regression analysis of sweat secretion volumes in cystic fibrosis patients, 23rd IEEE FRUCT Conference, Bologna 13-16 November 2018, Italy
• Lotti V, Kleinfelder K, Farinazzo A et al. Response to ivacaftor of the rare CFTR variants W57G and A234D in intestinal organoids and Fisher Rat Thyroid (FRT) cells, 43rd European Cystic Fibrosis Conference – 2020
• Sorio C, Nuovi approcci per la caratterizzazione funzionale di mutazioni in FC, 15th Meeting Nazionale della Società Italiana per lo Studio della Fibrosi Cistica (SIFC), 3-4 May 2019 – Rimini, Italy
• Sorio C, Potentiators and Correctors Therapy in Cystic Fibrosis, XLIII Congresso Nazionale Associazione Italiana per lo Studio del Pancreas (AISP), 19-21 Settembre 2019 – Verona, Italy
• Sorio C, CFF Theratyping Group, Lotti V et al. CFTR modulator theratyping using intestinal organoids from a CF patient and Fisher Rat Thyroid (FRT) cells expressing the rare CFTR variants W57G and A234D, Basic Science ECFS Conference 2020
• Sorio C, Lotti V, Kleinfelder K et al. Functional impact of a rare/varying clinical consequence CFTR variants W57G/A234D CFTR genotype and theratyping using rectal organoids, AMP Europe 2020 – Clinical Genomics: Beyond the Somatic Mutation