The most frequent CFTR mutations are deeply studied, while CFTR function and relation with clinical status remain poorly understood for the large group of “orphan” mutations. This project aims at in vitro expanding airway epithelial stem cells (AESC), derived from nasal epithelia of cystic fibrosis (CF) patients with different mutated genotypes, through the “culture reprogramming condition” (CRC) methodology. CF-CRC cultures will be validated for airway stem cell phenotype. Response to CFTR targeting drugs will be assessed in both AESC cells and AESC-derived organoids, through functional assays, and correlated to CFTR genetic profile, with the aim to predict the patient response. Finally, the specific CFTR defects will be replaced with wild type sequence in CF-CRC, through a combination of SFHR and Crispr/Cas9 gene editing approaches. The final aim of the project is to develop a new cellular epithelial model and a new source of organoids, virtually derived from each CF patient, where new drugs and even gene therapy may be tested.