@article {Prins631614, author = {Stella Prins and Emily Langron and Cato Hastings and Emily Hill and Andra C. Stefan and Lewis D. Griffin and Paola Vergani}, title = {High-content assay for precision medicine discovery in cystic fibrosis}, elocation-id = {631614}, year = {2019}, doi = {10.1101/631614}, publisher = {Cold Spring Harbor Laboratory}, abstract = {Cystic fibrosis (CF) is a life-limiting disease caused by mutations in the CFTR gene, which encodes an anion-selective channel. Because CF-causing mutations can affect CFTR gating as well as its biogenesis, multi-assay approaches have been implemented for drug development, to sequentially screen for channel function and membrane density. Here we present the first assay capable of simultaneous assessment of both CFTR characteristics.To validate our assay, we investigate F508del-CFTR, the most common disease-causing CFTR mutant, confirming rescue by incubation at low temperature, treatment with CFTR-targeting drugs and introduction of the second-site revertant mutation R1070W. In addition, we characterize a panel of rare CF-causing mutations and profile effects of acute treatment with approved VX-770 drug. Mapping the potentiation profile on CFTR structures reveals unprecedented mechanistic hypotheses on drug action.Simultaneous monitoring of CFTR localization and function can better guide CF drug development, beyond the artificial distinction between corrector and potentiator drugs. By providing a two-dimensional molecular characterization of individual mutant CFTR proteins, our assay is a powerful tool for development of therapies truly addressing the root cause of each individual patient{\textquoteright}s CF disease.}, URL = {https://www.biorxiv.org/content/early/2019/05/30/631614}, eprint = {https://www.biorxiv.org/content/early/2019/05/30/631614.full.pdf}, journal = {bioRxiv} }