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Updated: Aug 12, 2025
Forskolin-induced Swelling in Intestinal Organoids: An In Vitro Assay for Assessing Drug Response in Cystic Fibrosis Patients
Published on: February 11, 2017
High-throughput functional assay in cystic fibrosis patient-derived organoids allows drug repurposing
Sacha Spelier1,2,3, Eyleen de Poel1,2,3, Georgia N Ithakisiou1,2
1Department of Pediatric Respiratory Medicine, Wilhelmina Children's Hospital, University Medical Center, Utrecht University, Utrecht, The Netherlands.
Background:
Cystic fibrosis (CF) is a rare hereditary disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Recent therapies enable effective restoration of CFTR function of the most common F508del CFTR mutation. This shifts the unmet clinical need towards people with rare CFTR mutations such as nonsense mutations, of which G542X and W1282X are most prevalent. CFTR function measurements in patient-derived cell-based assays played a critical role in preclinical drug development for CF and may play an important role to identify new drugs for people with rare CFTR mutations.
Methods:
Here, we miniaturised the previously described forskolin-induced swelling (FIS) assay in intestinal organoids from a 96-well to a 384-well plate screening format. Using this novel assay, we tested CFTR increasing potential of a 1400-compound Food and Drug Administration (FDA)-approved drug library in organoids from donors with W1282X/W1282X CFTR nonsense mutations.
Results:
The 384-well FIS assay demonstrated uniformity and robustness based on coefficient of variation and Z'-factor calculations. In the primary screen, CFTR induction was limited overall, yet interestingly, the top five compound combinations that increased CFTR function all contained at least one statin. In the secondary screen, we indeed verified that four out of the five statins (mevastatin, lovastatin, simvastatin and fluvastatin) increased CFTR function when combined with CFTR modulators. Statin-induced CFTR rescue was concentration-dependent and W1282X-specific.
Conclusions:
Future studies should focus on elucidating genotype specificity and mode-of-action of statins in more detail. This study exemplifies proof of principle of large-scale compound screening in a functional assay using patient-derived organoids.
Insights
Statins show potential in rescuing CFTR function for rare mutations like W1282X in cystic fibrosis (CF) patients. This study used a miniaturized assay for large-scale drug screening in patient-derived organoids.
Area of Science:
- Biomedical research
- Genetics
- Pharmacology
Background:
- Cystic fibrosis (CF) is a genetic disorder caused by mutations in the CFTR gene.
- Current therapies target common CFTR mutations, leaving a need for treatments for rare mutations.
- Nonsense mutations, such as G542X and W1282X, represent a significant unmet need in CF care.
Purpose of the Study:
- To develop a miniaturized, high-throughput screening assay for CFTR function.
- To identify FDA-approved drugs that can restore function in CFTR nonsense mutations.
- To investigate the potential of statins in treating rare CFTR mutations.
Main Methods:
- Miniaturization of the forskolin-induced swelling (FIS) assay to a 384-well plate format.
- Screening of a 1400-compound FDA-approved drug library using intestinal organoids from W1282X/W1282X CF patients.
- Validation of identified compounds in secondary screens and dose-response studies.
Main Results:
- The 384-well FIS assay demonstrated robustness and uniformity.
- Combinations including statins were most effective in increasing CFTR function.
- Four statins (mevastatin, lovastatin, simvastatin, fluvastatin) were confirmed to enhance CFTR function in W1282X organoids.
- Statin-induced CFTR rescue was concentration-dependent and specific to the W1282X mutation.
Conclusions:
- Statins hold promise for treating specific rare CFTR mutations, particularly W1282X.
- This study provides a proof-of-principle for large-scale drug screening using patient-derived organoids.
- Further research is needed to explore the precise mechanism of action and genotype specificity of statins in CF.
