A functional CFTR assay using primary cystic fibrosis intestinal organoids

J.F. Dekkers, C.L. Wiegerinck, H.R. de Jonge, I. Bronsveld, H.M. Janssens, K.M. de Winter-de Groot, A.M. Brandsma, N.W. de Jong, M.J. Bijvelds, B.J. Scholte, E.E. Nieuwenhuis, S. van den Brink, H. Clevers, C.K. van der Ent, S. Middendorp, J.M. Beekman

Research output: Contribution to journal/periodicalArticleScientificpeer-review

Abstract

We recently established conditions allowing for long-term expansion of epithelial organoids from intestine, recapitulating essential features of the in vivo tissue architecture. Here we apply this technology to study primary intestinal organoids of people suffering from cystic fibrosis, a disease caused by mutations in CFTR, encoding cystic fibrosis transmembrane conductance regulator. Forskolin induces rapid swelling of organoids derived from healthy controls or wild-type mice, but this effect is strongly reduced in organoids of subjects with cystic fibrosis or in mice carrying the Cftr F508del mutation and is absent in Cftr-deficient organoids. This pattern is phenocopied by CFTR-specific inhibitors. Forskolin-induced swelling of in vitro-expanded human control and cystic fibrosis organoids corresponds quantitatively with forskolin-induced anion currents in freshly excised ex vivo rectal biopsies. Function of the CFTR F508del mutant protein is restored by incubation at low temperature, as well as by CFTR-restoring compounds. This relatively simple and robust assay will facilitate diagnosis, functional studies, drug development and personalized medicine approaches in cystic fibrosis.
Original languageEnglish
Pages (from-to)939-945
JournalNature Medicine
Volume19
Issue number7
DOIs
Publication statusPublished - 2013

Fingerprint Dive into the research topics of 'A functional CFTR assay using primary cystic fibrosis intestinal organoids'. Together they form a unique fingerprint.

  • Cite this

    Dekkers, J. F., Wiegerinck, C. L., de Jonge, H. R., Bronsveld, I., Janssens, H. M., de Winter-de Groot, K. M., Brandsma, A. M., de Jong, N. W., Bijvelds, M. J., Scholte, B. J., Nieuwenhuis, E. E., van den Brink, S., Clevers, H., van der Ent, C. K., Middendorp, S., & Beekman, J. M. (2013). A functional CFTR assay using primary cystic fibrosis intestinal organoids. Nature Medicine, 19(7), 939-945. https://doi.org/10.1038/nm.3201