Anabela S Ramalho1, Eva Fürstová2, Annelotte M Vonk3,4, Marc Ferrante5,6, Catherine Verfaillie7, Lieven Dupont8,9, Mieke Boon1,10, Marijke Proesmans1,10, Jeffrey M Beekman3,4, Ifat Sarouk11, Carlos Vazquez Cordero12, Francois Vermeulen1,10, Kris De Boeck1,10. 1. Dept of Development and Regeneration, Woman and Child Unit, CF Research Lab, KU Leuven, Leuven, Belgium. 2. Dept of Pediatrics, 2nd Faculty of Medicine, Charles University and Motol University Hospital, Prague, Czech Republic. 3. Dept of Pediatric Pulmonology, Wilhelmina Children's Hospital, University Medical Centre, Utrecht, The Netherlands. 4. Regenerative Medicine Center, University Medical Centre, Utrecht, The Netherlands. 5. Dept of Chronic Diseases, Metabolism and Ageing, Translational Research Center for Gastrointestinal Disorders (TARGID), KU Leuven, Leuven, Belgium. 6. Dept of Gastroenterology and Hepatology, University Hospitals Leuven, Leuven, Belgium. 7. Dept of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven, Leuven, Belgium. 8. Dept of Chronic Diseases, Metabolism and Ageing; Pneumology, KU Leuven, Leuven, Belgium. 9. Dept of Respiratory Diseases, University Hospital of Leuven, Leuven, Belgium. 10. Dept of Pediatrics, Pediatric Pulmonology, University Hospital of Leuven, Leuven, Belgium. 11. Pulmonology Pediatrics and National CF Center Edmond and Lily Safra Children's Hospital, Sheba Medical Center, Tel Hashomer, Israel. 12. Pulmonology and Cystic Fibrosis Unit, Cruces University Hospital, Barakaldo, Spain.
Abstract
RATIONALE: Given the vast number of cystic fibrosis transmembrane conductance regulator (CFTR) mutations, biomarkers predicting benefit from CFTR modulator therapies are needed for subjects with cystic fibrosis (CF). OBJECTIVES: To study CFTR function in organoids of subjects with common and rare CFTR mutations and evaluate correlations between CFTR function and clinical data. METHODS: Intestinal organoids were grown from rectal biopsies in a cohort of 97 subjects with CF. Residual CFTR function was measured by quantifying organoid swelling induced by forskolin and response to modulators by quantifying organoid swelling induced by CFTR correctors, potentiator and their combination. Organoid data were correlated with clinical data from the literature. RESULTS: Across 28 genotypes, residual CFTR function correlated (r2=0.87) with sweat chloride values. When studying the same genotypes, CFTR function rescue by CFTR modulators in organoids correlated tightly with mean improvement in lung function (r2=0.90) and sweat chloride (r2=0.95) reported in clinical trials. We identified candidate genotypes for modulator therapy, such as E92K, Q237E, R334W and L159S. Based on organoid results, two subjects started modulator treatment: one homozygous for complex allele Q359K_T360K, and the second with mutation E60K. Both subjects had major clinical benefit. CONCLUSIONS: Measurements of residual CFTR function and rescue of function by CFTR modulators in intestinal organoids correlate closely with clinical data. Our results for reference genotypes concur with previous results. CFTR function measured in organoids can be used to guide precision medicine in patients with CF, positioning organoids as a potential in vitro model to bring treatment to patients carrying rare CFTR mutations.
RATIONALE: Given the vast number of cystic fibrosis transmembrane conductance regulator (CFTR) mutations, biomarkers predicting benefit from CFTR modulator therapies are needed for subjects with cystic fibrosis (CF). OBJECTIVES: To study CFTR function in organoids of subjects with common and rare CFTR mutations and evaluate correlations between CFTR function and clinical data. METHODS: Intestinal organoids were grown from rectal biopsies in a cohort of 97 subjects with CF. Residual CFTR function was measured by quantifying organoid swelling induced by forskolin and response to modulators by quantifying organoid swelling induced by CFTR correctors, potentiator and their combination. Organoid data were correlated with clinical data from the literature. RESULTS: Across 28 genotypes, residual CFTR function correlated (r2=0.87) with sweat chloride values. When studying the same genotypes, CFTR function rescue by CFTR modulators in organoids correlated tightly with mean improvement in lung function (r2=0.90) and sweat chloride (r2=0.95) reported in clinical trials. We identified candidate genotypes for modulator therapy, such as E92K, Q237E, R334W and L159S. Based on organoid results, two subjects started modulator treatment: one homozygous for complex allele Q359K_T360K, and the second with mutation E60K. Both subjects had major clinical benefit. CONCLUSIONS: Measurements of residual CFTR function and rescue of function by CFTR modulators in intestinal organoids correlate closely with clinical data. Our results for reference genotypes concur with previous results. CFTR function measured in organoids can be used to guide precision medicine in patients with CF, positioning organoids as a potential in vitro model to bring treatment to patients carrying rare CFTR mutations.
Authors: Sharon L Wong; Nikhil T Awatade; Miro A Astore; Katelin M Allan; Michael J Carnell; Iveta Slapetova; Po-Chia Chen; Jeffry Setiadi; Elvis Pandzic; Laura K Fawcett; John R Widger; Renee M Whan; Renate Griffith; Chee Y Ooi; Serdar Kuyucak; Adam Jaffe; Shafagh A Waters Journal: Am J Respir Cell Mol Biol Date: 2022-07 Impact factor: 7.748
Authors: Justin D Anderson; Zhongyu Liu; L Victoria Odom; Latona Kersh; Jennifer S Guimbellot Journal: Am J Physiol Lung Cell Mol Physiol Date: 2021-05-19 Impact factor: 6.011
Authors: Giovana B Bampi; Anabela S Ramalho; Leonardo A Santos; Johannes Wagner; Lieven Dupont; Harry Cuppens; Kris De Boeck; Zoya Ignatova Journal: Life (Basel) Date: 2020-12-27