TY - JOUR
T1 - R117H-CFTR function and response to VX-770 correlate with mRNA and protein expression in intestinal organoids
AU - Van Mourik, Peter
AU - van Haaren, Paul
AU - Kruisselbrink, Evelien
AU - Korkmaz, Cemil
AU - Janssens, Hettie M.
AU - de Winter – de Groot, Karin M.
AU - van der Ent, Cornelis K.
AU - Hagemeijer, Marne C.
AU - Beekman, Jeffrey M.
N1 - Funding Information:
This work was supported by grants of the Dutch Cystic Fibrosis Foundation (NCFS, HIT-CF grant) and Netherlands Organization for Health Research and Development (ZonMw). Funding sources had no involvement in study design, collection, analysis, interpretation or writing.
Funding Information:
Mouse ?-CFTR antibodies 450, 570, 596 were provided by the Cystic Fibrosis Folding Consortium and the rabbit ?-HSP90 antibody was a kind gift from prof. Dr. Ineke Braakman from Utrecht University.
Publisher Copyright:
© 2020 European Cystic Fibrosis Society
PY - 2020/9
Y1 - 2020/9
N2 - Introduction: Variability in disease severity and CFTR modulator responses exists between patients with identical CFTR genotypes. Here, we characterized transcription, translation and function of R117H-CFTR using intestinal organoids and correlated them with in vitro responses to ivacaftor (VX-770). Methods: Organoids were generated from individuals possessing at least one R117H-CFTR allele. The forskolin-induced swelling (FIS) assay was used to measure CFTR function and response to VX-770 treatment. R117H-CFTR protein and mRNA expression levels were determined in parallel and Pearson's correlation coefficients were assessed. Results: Variability in R117H-CFTR FIS responses was observed and correlated significantly with mRNA and protein expression. Response to VX-770 treatment in organoids correlated with mRNA and protein expression as well. Conclusions: Our results indicate that gene expression, protein expression and CFTR function are strongly correlated in organoids from people with CFTR-R117H-7T/9T, which may suggest that CFTR gene expression may have consequences for CF diagnosis, prognosis and therapeutic benefit.
AB - Introduction: Variability in disease severity and CFTR modulator responses exists between patients with identical CFTR genotypes. Here, we characterized transcription, translation and function of R117H-CFTR using intestinal organoids and correlated them with in vitro responses to ivacaftor (VX-770). Methods: Organoids were generated from individuals possessing at least one R117H-CFTR allele. The forskolin-induced swelling (FIS) assay was used to measure CFTR function and response to VX-770 treatment. R117H-CFTR protein and mRNA expression levels were determined in parallel and Pearson's correlation coefficients were assessed. Results: Variability in R117H-CFTR FIS responses was observed and correlated significantly with mRNA and protein expression. Response to VX-770 treatment in organoids correlated with mRNA and protein expression as well. Conclusions: Our results indicate that gene expression, protein expression and CFTR function are strongly correlated in organoids from people with CFTR-R117H-7T/9T, which may suggest that CFTR gene expression may have consequences for CF diagnosis, prognosis and therapeutic benefit.
KW - CFTR
KW - mRNA
KW - Organoids
KW - R117H
KW - VX-770
UR - http://www.scopus.com/inward/record.url?scp=85079267738&partnerID=8YFLogxK
U2 - 10.1016/j.jcf.2020.02.001
DO - 10.1016/j.jcf.2020.02.001
M3 - Article
AN - SCOPUS:85079267738
SN - 1569-1993
VL - 19
SP - 728
EP - 732
JO - Journal of Cystic Fibrosis
JF - Journal of Cystic Fibrosis
IS - 5
ER -