TY - JOUR
T1 - Measurements of functional responses in human primary lung cells as a basis for personalized therapy for cystic fibrosis
AU - Awatade, Nikhil T.
AU - Uliyakina, Inna
AU - Farinha, Carlos M.
AU - Clarke, Luka A.
AU - Mendes, Ana
AU - Solé, Amparo
AU - Pastor, Juan
AU - Ramos, Maria Margarida
AU - Amaral, Margarida D.
N1 - Funding Information:
This work was supported by PEst-OE/BIA/UI4046/2011 ( POCTI/FCT/PIDDAC ), POCTI/PTDC/SAU-GMG/122229/2010 ( FCT , Portugal) and Gilead GÉNESE Programme grant (Ref MED-2013-250 ) and IU are recipients of doctoral fellowships SFRH/BD/52487/2014 and SFRH/BD/69180/2010 , respectively from FCT , Portugal. The authors thank Professor Bob Bridges (RFUMs, Chicago, IL, USA) and CFF (USA) for supplying compound C18 and Inh 172 and are also grateful to Ines Pérez and Salvador Bejar Carbonell from Hospital la Fe (Valencia, Spain) for help with handling and shipping of patients' materials and to Verónica Felício (FCUL, Lisboa) for help in the establishment of primary cultures.
Funding Information:
MDA has served as a consultant to Vertex and Galapagos, and has been supported to attend and to speak at Symposia (Novartis, Gilead and Vertex) and to participate in an educational grant programme by Facilitate Ltd. AS has served as a consultant to Vertex.
Publisher Copyright:
© 2015.
PY - 2015/2
Y1 - 2015/2
N2 - Background: The best investigational drug to treat cystic fibrosis (CF) patients with the most common CF-causing mutation (F508del) is VX-809 (lumacaftor) which recently succeeded in Phase III clinical trial in combination with ivacaftor. This corrector rescues F508del-CFTR from its abnormal intracellular localization to the cell surface, a traffic defect shared by all Class II CFTR mutants. Our goal here is to test the efficacy of lumacaftor in other Class II mutants in primary human bronchial epithelial (HBE) cells derived from CF patients. Methods: The effect of lumacaftor was investigated in primary HBE cells from non-CF and CF patients with F508del/F508del, A561E/A561E, N1303K/G542X, F508del/G542X and F508del/Y1092X genotypes by measurements of Forskolin plus Genistein-inducible equivalent short-circuit current (Ieq-SC-Fsk+Gen) in perfused open-circuit Ussing chambers. Efficacy of corrector C18 was also assessed on A561E/A561E and F508del/F508del cells. Results: Our data indicate that A561E (when present in both alleles) responds positively to lumacaftor treatment at equivalent efficacy of F508del in primary HBE cells. Similarly, lumacaftor has a positive impact on Y1092X, but not on N1303K. Our data also show that cells with only one copy of F508del-CFTR respond less to VX-809. Moreover, there is great variability in lumacaftor responses among F508del-homozygous cells from different donors. Compound C18 failed to rescue A561E-CFTR but not in F508del-CFTR, thus plausibly it has a different mechanism of action distinct from lumacaftor. Conclusions: CF patients with A561E (and likely also those with Y1029X) can potentially benefit from lumacaftor. Moreover, the methodology used here exemplifies how ex vivo approaches may apply personalized therapies to CF and possibly other respiratory diseases.
AB - Background: The best investigational drug to treat cystic fibrosis (CF) patients with the most common CF-causing mutation (F508del) is VX-809 (lumacaftor) which recently succeeded in Phase III clinical trial in combination with ivacaftor. This corrector rescues F508del-CFTR from its abnormal intracellular localization to the cell surface, a traffic defect shared by all Class II CFTR mutants. Our goal here is to test the efficacy of lumacaftor in other Class II mutants in primary human bronchial epithelial (HBE) cells derived from CF patients. Methods: The effect of lumacaftor was investigated in primary HBE cells from non-CF and CF patients with F508del/F508del, A561E/A561E, N1303K/G542X, F508del/G542X and F508del/Y1092X genotypes by measurements of Forskolin plus Genistein-inducible equivalent short-circuit current (Ieq-SC-Fsk+Gen) in perfused open-circuit Ussing chambers. Efficacy of corrector C18 was also assessed on A561E/A561E and F508del/F508del cells. Results: Our data indicate that A561E (when present in both alleles) responds positively to lumacaftor treatment at equivalent efficacy of F508del in primary HBE cells. Similarly, lumacaftor has a positive impact on Y1092X, but not on N1303K. Our data also show that cells with only one copy of F508del-CFTR respond less to VX-809. Moreover, there is great variability in lumacaftor responses among F508del-homozygous cells from different donors. Compound C18 failed to rescue A561E-CFTR but not in F508del-CFTR, thus plausibly it has a different mechanism of action distinct from lumacaftor. Conclusions: CF patients with A561E (and likely also those with Y1029X) can potentially benefit from lumacaftor. Moreover, the methodology used here exemplifies how ex vivo approaches may apply personalized therapies to CF and possibly other respiratory diseases.
KW - Innovative treatments
KW - Mutation-specific therapies
KW - Personalized medicine
KW - Rare diseases
UR - http://www.scopus.com/inward/record.url?scp=84943230777&partnerID=8YFLogxK
U2 - 10.1016/j.ebiom.2014.12.005
DO - 10.1016/j.ebiom.2014.12.005
M3 - Article
C2 - 26137539
AN - SCOPUS:84943230777
SN - 2352-3964
VL - 2
SP - 147
EP - 153
JO - EBioMedicine
JF - EBioMedicine
IS - 2
ER -