ABCMdb

PMID: 14534336

Wilschanski M, Yahav Y, Yaacov Y, Blau H, Bentur L, Rivlin J, Aviram M, Bdolah-Abram T, Bebok Z, Shushi L, Kerem B, Kerem E
Gentamicin-induced correction of CFTR function in patients with cystic fibrosis and CFTR stop mutations.
N Engl J Med. 2003 Oct 9;349(15):1433-41., 2003-10-09 [PubMed]

Sentences

No. Mutations Sentence Comment

15 ABCC7 p.Trp1282* ABCC7 p.Trp1282* ABCC7 p.Gly542* Howardetal.demonstratedthattwoCFTR-associated stop mutations could be suppressed by treating cells with low doses of an aminoglycoside antibiotic.12 Bedwell et al. demonstrated that after incubation of bronchial epithelial cell line IB3-1, whichcarriesaW1282XmutationofCFTR,withami- noglycosides, cyclic AMP (cAMP)-activated chloride conductance and the expression of functional CFTR were restored to the apical membrane.13 Recently, Zsembery et al. isolated cholangiocytes from the liver of a patient carrying the G542X mutation of CFTR and incubated them with gentamicin, resulting in the expression of cAMP-activated chloride transport.14 Thus, in vitro, gentamicin obviated the effect of stop-codon mutations on the transcription and translation of CFTR. This effect has subsequently been demonstrated in a number of models of other diseases caused by stop mutations,includingmusculardystrophy,15 Hurler`ssyndrome,16 cystinosis,17 late infantile neuronal ceroid lipofuscinosis,18 and disorders involving the p53 gene.19 In a previous open pilot study, we found that topical application of gentamicin drops to the nose augmented chloride transport in epithelial cells of nine patients with cystic fibrosis who had at least one W1282X allele.20 Subsequently, Clancy et al.,21 in an open study, administered gentamicin intravenously to five patients who were heterozygous for stop mutations and found that four of the patients had hyperpolarization of the nasal potential differ- enceaftertheadministrationofisoproterenol,indicating that chloride transport was induced across the apical surface. Login to comment 34 ABCC7 p.Trp1282* immunofluorescence microscopy of primary human airway cells Primarynasalepithelialcellsfromtwopatientswith cystic fibrosis who were compound heterozygotes for the W1282X and the ∆F508 mutations and who participated in the gentamicin study were obtained by scraping before and after treatment with gentamicin and were then spread on microscope slides. Login to comment 62 ABCC7 p.Trp1282* ABCC7 p.Trp1282* ABCC7 p.Trp1282* ABCC7 p.Gly542* Of the 24 study patients,11carriedtwostopmutations:6werehomo- zygous for W1282X, 3 were compound heterozygous for W1282X/G542X, and 2 were compound heterozygous for W1282X/3849+10KbC˚T. Login to comment 63 ABCC7 p.Trp1282* ABCC7 p.Trp1282* ABCC7 p.Trp1282* ABCC7 p.Gly85Glu The 3849+10kbC˚T mutation can lead to the inclusion of a cryptic 84-bp exon, which contains a stop codon.24 Another eight patients were heterozygous for stop mutations: six were ∆F508/W1282X, one was G85E/ W1282X, and one was W1282X/unknown. Login to comment 89 ABCC7 p.Trp1282* detection of full-length cftr protein The effect of intranasal gentamicin treatment on the "read through" of premature nonsense codons was further analyzed in two of the patients with the ∆F508/W1282X genotype who had a response to gentamicin. Login to comment 107 ABCC7 p.Trp1282* Furthermore, staining the cells with an antibody against CFTR, which recognizes a region in CFTR beyond the W1282X mutation, showed an increase in the numberofcellswithsurfacestaining,indicatingthe delivery of full-length CFTR proteins to the apical membrane. Login to comment 108 ABCC7 p.Trp1282* These results provide direct evidence that gentamicin treatment of patients with cystic fibrosis and the stop mutation W1282X can induce the synthesis of full-length CFTR. This correction of the abnormal potential difference and the appearance of full-length CFTR on the cell surface support the findings of previous studies,whichshowedthatgentamicincanpromote "read through" of stop mutations. Login to comment 117 ABCC7 p.Trp1282* An Example of Immunostaining for the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) in a Negative Control Involving Nonimmune Mouse CFTR IgG (Panel A) and in a Patient with the W1282X/∆∆∆∆F508 Mutation before (Panel B) and after (Panel C) Gentamicin Treatment (¬100). Login to comment 129 ABCC7 p.Arg553* ABCC7 p.Gly542* In vitro studies using quantitative immunohistochemistry have shown that after incubation with an aminoglycoside, cells from patients with cystic fibrosis have as much as 25 percent (in those with the R553X mutation) to 35 percent (in those with the G542X mutation) of the concentration of full-length CFTR observed in cells transfected with a wild-type CFTR complementary DNA.12,13 This in- creaseinfunctionalCFTRmightexceedthethresh- old required for normally functioning respiratory epithelial cells and might thus have corrected cell-membrane function in our patients. Login to comment 158 ABCC7 p.Trp1282* ShoshaniT,AugartenA,GazitE,etal.Association of a nonsense mutation (W1282X), themostcommonmutationintheAshkena- zi Jewish cystic fibrosis patients in Israel, with presentation of severe disease. Login to comment