PMID- 23727931
OWN - NLM
STAT- MEDLINE
DCOM- 20130923
LR  - 20220317
IS  - 1546-170X (Electronic)
IS  - 1078-8956 (Linking)
VI  - 19
IP  - 7
DP  - 2013 Jul
TI  - A functional CFTR assay using primary cystic fibrosis intestinal organoids.
PG  - 939-45
LID - 10.1038/nm.3201 [doi]
AB  - 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.
FAU - Dekkers, Johanna F
AU  - Dekkers JF
AD  - Department of Pediatric Pulmonology, Wilhelmina Children's Hospital, University 
      Medical Center, Utrecht, The Netherlands.
FAU - Wiegerinck, Caroline L
AU  - Wiegerinck CL
FAU - de Jonge, Hugo R
AU  - de Jonge HR
FAU - Bronsveld, Inez
AU  - Bronsveld I
FAU - Janssens, Hettie M
AU  - Janssens HM
FAU - de Winter-de Groot, Karin M
AU  - de Winter-de Groot KM
FAU - Brandsma, Arianne M
AU  - Brandsma AM
FAU - de Jong, Nienke W M
AU  - de Jong NW
FAU - Bijvelds, Marcel J C
AU  - Bijvelds MJ
FAU - Scholte, Bob J
AU  - Scholte BJ
FAU - Nieuwenhuis, Edward E S
AU  - Nieuwenhuis EE
FAU - van den Brink, Stieneke
AU  - van den Brink S
FAU - Clevers, Hans
AU  - Clevers H
FAU - van der Ent, Cornelis K
AU  - van der Ent CK
FAU - Middendorp, Sabine
AU  - Middendorp S
FAU - Beekman, Jeffrey M
AU  - Beekman JM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130602
PL  - United States
TA  - Nat Med
JT  - Nature medicine
JID - 9502015
RN  - 0 (Mutant Proteins)
RN  - 059QF0KO0R (Water)
RN  - 126880-72-6 (Cystic Fibrosis Transmembrane Conductance Regulator)
SB  - IM
MH  - Animals
MH  - Biological Assay/methods
MH  - Cells, Cultured
MH  - Cystic Fibrosis/diagnosis/genetics/*pathology
MH  - Cystic Fibrosis Transmembrane Conductance Regulator/*analysis/genetics/metabolism
MH  - Female
MH  - HEK293 Cells
MH  - Humans
MH  - Intestinal Mucosa/metabolism/*pathology
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mice, Inbred CFTR
MH  - Mutant Proteins/analysis/genetics/metabolism/physiology
MH  - Organoids/metabolism/*pathology
MH  - Primary Cell Culture/methods
MH  - Water/metabolism
EDAT- 2013/06/04 06:00
MHDA- 2013/09/24 06:00
CRDT- 2013/06/04 06:00
PHST- 2012/01/20 00:00 [received]
PHST- 2012/12/20 00:00 [accepted]
PHST- 2013/06/04 06:00 [entrez]
PHST- 2013/06/04 06:00 [pubmed]
PHST- 2013/09/24 06:00 [medline]
AID - nm.3201 [pii]
AID - 10.1038/nm.3201 [doi]
PST - ppublish
SO  - Nat Med. 2013 Jul;19(7):939-45. doi: 10.1038/nm.3201. Epub 2013 Jun 2.