Journal of Molecular Medicine

, 89:1149

Suppression of CFTR premature termination codons and rescue of CFTR protein and function by the synthetic aminoglycoside NB54

  • Steven M. Rowe
  • Peter Sloane
  • Li Ping Tang
  • Kyle Backer
  • Marina Mazur
  • Jessica Buckley-Lanier
  • Igor Nudelman
  • Valery Belakhov
  • Zsuzsa Bebok
  • Erik Schwiebert
  • Timor Baasov
  • David M. Bedwell
Original Article

DOI: 10.1007/s00109-011-0787-6

Cite this article as:
Rowe, S.M., Sloane, P., Tang, L.P. et al. J Mol Med (2011) 89: 1149. doi:10.1007/s00109-011-0787-6

Abstract

Certain aminoglycosides are capable of inducing “translational readthrough” of premature termination codons (PTCs). However, toxicity and relative lack of efficacy deter treatment with clinically available aminoglycosides for genetic diseases caused by PTCs, including cystic fibrosis (CF). Using a structure-based approach, the novel aminoglycoside NB54 was developed that exhibits reduced toxicity and enhanced suppression of PTCs in cell-based reporter assays relative to gentamicin. We examined whether NB54 administration rescued CFTR protein and function in clinically relevant CF models. In a fluorescence-based halide efflux assay, NB54 partially restored halide efflux in a CF bronchial epithelial cell line (CFTR genotype W1282X/F508del), but not in a CF epithelial cell line lacking a PTC (F508del/F508del). In polarized airway epithelial cells expressing either a CFTR-W1282X or -G542X cDNA, treatment with NB54 increased stimulated short-circuit current (ISC) with greater efficiency than gentamicin. NB54 and gentamicin induced comparable increases in forskolin-stimulated ISC in primary airway epithelial cells derived from a G542X/F508del CF donor. Systemic administration of NB54 to Cftr−/− mice expressing a human CFTR-G542X transgene restored 15–17% of the average stimulated transepithelial chloride currents observed in wild-type (Cftr+/+) mice, comparable to gentamicin. NB54 exhibited reduced cellular toxicity in vitro and was tolerated at higher concentrations than gentamicin in vivo. These results provide evidence that synthetic aminoglycosides are capable of PTC suppression in relevant human CF cells and a CF animal model and support further development of these compounds as a treatment modality for genetic diseases caused by PTCs.

Keywords

Cystic fibrosis transmembrane conductance regulator Premature termination codons Aminoglycosides Short-circuit current Primary human bronchial epithelial cells 

Abbreviations

AHB

(S)-4-Amino-2 hydroxybutanoyl

CF

Cystic fibrosis

CFTR

Cystic fibrosis transmembrane conductance regulator

ISC

Short-circuit current

PTC

Premature termination codons

SPQ

6-Methoxy-N-(3-sulfopropyl)-quinolinium

Supplementary material

109_2011_787_MOESM1_ESM.pdf (47 kb)
ESM 1PDF 46 kb

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Steven M. Rowe
    • 1
    • 2
    • 3
    • 6
    • 9
  • Peter Sloane
    • 1
    • 6
  • Li Ping Tang
    • 1
    • 6
  • Kyle Backer
    • 1
    • 6
  • Marina Mazur
    • 6
  • Jessica Buckley-Lanier
    • 4
  • Igor Nudelman
    • 7
  • Valery Belakhov
    • 7
  • Zsuzsa Bebok
    • 5
    • 6
  • Erik Schwiebert
    • 8
  • Timor Baasov
    • 7
  • David M. Bedwell
    • 3
    • 4
    • 6
  1. 1.Department of MedicineUniversity of Alabama at BirminghamBirminghamUSA
  2. 2.Department of PediatricsUniversity of Alabama at BirminghamBirminghamUSA
  3. 3.Department of Physiology and BiophysicsUniversity of Alabama at BirminghamBirminghamUSA
  4. 4.Department of MicrobiologyUniversity of Alabama at BirminghamBirminghamUSA
  5. 5.Department of Cell BiologyUniversity of Alabama at BirminghamBirminghamUSA
  6. 6.Gregory Fleming James Cystic Fibrosis Research CenterUniversity of Alabama at BirminghamBirminghamUSA
  7. 7.The Edith and Joseph Fisher Enzyme Inhibitors Laboratory, Schulich Faculty of ChemistryTechnion-Israel Institute of TechnologyHaifaIsrael
  8. 8.DiscoveryBioMed, Inc.BirminghamUSA
  9. 9.University of Alabama at BirminghamBirminghamUSA

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