Patient-derived cell models and genome engineering for precision therapy of cystic fibrosis

 Cystic fibrosis (CF) is the most common life-shortening monogenic disorder causing multi-organ disease. CF is caused by mutations in the CFTR gene, which encodes a Cl-/HCO3- channel expressed in the apical membrane of epithelial cells. Current small molecule treatment is available to ~60% of CF patients and with new modulators in the pipeline, potentially 90% of CF patients can be treated. The last 10%, i.e. the minimal function mutations, remain without treatment and have been poorly characterized to date. To obtain highest translational value, studying CFTR subcellular localization, processing and trafficking in (near-)physiological models is a prerequisite, but due to low expression of endogenous CFTR and absence of specific and sensitive antibodies, this remains challenging. My aim is to generate innovative cell models and assays to deepen our basic knowledge on physiological CFTR expression, trafficking and function by increasing: (1) the sensitivity of detecting endogenous CFTR and (2) the resolution to study cellular defects of mutations at single cell level. I will combine relevant primary cell models (intestinal organoids, airway cultures) with viral vectors, CRISPR/Cas and micro-chip technology from Imec to generate these novel assays. They will allow me to characterize rare CFTR mutations in a physiological context and evaluate hit activity from a repurposing screen we recently performed for two such mutants, as a precision medicine strategy for CF.