ABCMdb

PMID: 23727931

Dekkers JF, Wiegerinck CL, de Jonge HR, Bronsveld I, Janssens HM, de Winter-de Groot KM, Brandsma AM, de Jong NW, Bijvelds MJ, Scholte BJ, Nieuwenhuis EE, van den Brink S, Clevers H, van der Ent CK, Middendorp S, Beekman JM
A functional CFTR assay using primary cystic fibrosis intestinal organoids.
Nat Med. 2013 Jul;19(7):939-45. doi: 10.1038/nm.3201. Epub 2013 Jun 2., [PubMed]

Sentences

No. Mutations Sentence Comment

16 ABCC7 p.Gly551Asp A phase 3 clinical trial has been successfully completed using the potentiator VX-770 (ivacaftor, Kalydeco) in subjects with the CFTR G551D mutation, demonstrating that mutation-specific drug targeting is feasible8. Login to comment 91 ABCC7 p.Glu60* We observed no FIS in E60X 4015delATTT organoids (Supplementary Video 2). Login to comment 121 ABCC7 p.Glu60* (a) Western blot analysis of CFTR and E-cadherin (loading control) expression in human rectal healthy control (HC; n = 2), E60X 4015delATTT (n = 1) and homozygous F508del CFTR (n = 2) organoids (top) and CFTR and ezrin (loading control) expression in whole-cell lysates of human rectal organoids that were either not treated (control) or treated with the deglycosylation enzyme Endo H or PNGase F (bottom). Login to comment 127 ABCC7 p.Ala455Glu ABCC7 p.Gly542* ABCC7 p.Glu60* ABCC7 p.Leu927Pro (d) Forskolin-induced swelling of rectal organoids derived from three individual healthy controls, two individuals with a mild cystic fibrosis genotype (F508del A455E) and nine individuals with a severe cystic fibrosis genotype (one individual with E60X 4015ATTTdel, one with F508del G542X, one with F508del L927P and six with F508del F508del). Login to comment 134 ABCC7 p.Gly542* ABCC7 p.Glu60* (Healthy controls, n = 3; mild cystic fibrosis, n = 2; severe cystic fibrosis (F508del F508del), n = 5; severe cystic fibrosis (other; E60X 4015ATTTdel and F508del G542X), n = 2; mean &#b1; s.d.). Login to comment 139 ABCC7 p.Glu60* All F508del compound heterozygous organoids also responded to correction, but no correction or potentiation was observed in E60X 4015delATTT organoids (Fig. 5a-c). Login to comment 140 ABCC7 p.Ala455Glu We next compared the efficacy of approaches to restore the function of F508del CFTR by comparing the amount of FIS to that in mock-treated F508del A455E organoids or healthy control organoids (Fig. 5d). Login to comment 141 ABCC7 p.Ala455Glu This comparison indicated that VX-809 is the most potent corrector and combined treatment with VX-809 and VX-770 induced FIS beyond the amounts in F508del A455E organoids, reaching ~60% of the FIS in healthy controls. Login to comment 150 ABCC7 p.Ala455Glu ABCC7 p.Gly542* ABCC7 p.Glu60* ABCC7 p.Leu927Pro 0 500 1,000 1,500 2,000 2,500 0 500 1,000 1,500 2,000 2,500 3,000 0 400 800 1,200 1,600 C8 Corr-4a C8 + Corr-4a VX-809 VX-770 VX-809 + VX-770 VRT-325 Corr-4a VRT-325 + Corr-4a CF1 CF6 CF5 CF4 CF3 CF2 F508del L927P F508del G542X E60X 4015delATTT F508del F508del b d a c 0 20 40 60 80 100 120 V R T - 3 2 5 C o r r - 4 a C 8 V X - 8 0 9 V X - 7 7 0 V R T - 3 2 5 + C o r r - 4 a C 8 + C o r r - 4 a V X - 8 0 9 + V X - 7 7 0 C o n t r o l C o n t r o l F508del F508del HC F508del A455E Organoid swelling (absolute AUC t = 60) Organoid swelling (absolute AUC t = 60) Organoid swelling (absolute AUC t = 60) Organoid swelling (normalized AUC t = 60) Figure 5ߒ Differential FIS between organoids from subjects with cystic fibrosis after chemical CFTR restoration. Login to comment 154 ABCC7 p.Ala455Glu (d) Average FIS responses of compound-treated F508del F508del organoids (n = 6 from a-c) and DMSO-treated F508del A455E organoids (n = 2) relative to the average FIS of DMSO-treated healthy control organoids (n = 3) expressed as the AUC calculated from the time periods shown in Figure 4d-f (baseline = 100%, t = 60 min; mean &#b1; s.e.m.). Login to comment 182 ABCC7 p.Ala455Glu This combination induces approximately 1.5-fold more FIS in CFTR F508del homozygous organoids as compared to untreated F508del A455E organoids and up to 60% of the FIS seen in healthy controls. Login to comment 245 ABCC7 p.Gly551Asp Ramsey, B.W. et al. A CFTR potentiator in patients with cystic fibrosis and the G551D mutation. Login to comment 332 ABCC7 p.Leu927Pro Hermans, C.J., Veeze, H.J., Drexhage, V.R., Halley, D.J. & van den Ouweland, A.M. Identification of the L927P and ࢞L1260 mutations in the CFTR gene. Login to comment 390 ABCC7 p.Gly542* ABCC7 p.Glu60* ABCC7 p.Leu927Pro Rectal organoids from healthy controls and subjects with cystic fibrosis were generated from four rectal suction biopsies after ICMs obtained (i) during standard cystic fibrosis care (one individual each harboring E60X 4015ATTTdel, F508del G542X or F508del L927P and five individuals harboring F508del F508del), (ii) for diagnostic purposes (one healthy control) or (iii) during voluntary participation in studies approved by the University Medical Center Utrecht and Erasmus MC ethics committees (two healthy controls and one individual harboring F508del F508del). Login to comment