Discovery of Clinically Approved Agents That Promote Suppression of Cystic Fibrosis Transmembrane Conductance Regulator Nonsense Mutations

Overview

Journal Am J Respir Crit Care Med

Specialty Critical Care

Date 2016 Apr 23

PMID 27104944

Citations 50

Authors

Venkateshwar Mutyam

Ming Du

Xiaojiao Xue

Kim M Keeling

E Lucile White

J Robert Bostwick

Lynn Rasmussen

Bo Liu

Marina Mazur

Jeong S Hong

Emily Falk Libby

Feng Liang

Haibo Shang

Martin Mense

Mark J Suto

David M Bedwell

Steven M Rowe

Affiliations

Soon will be listed here.

Abstract

Rationale: Premature termination codons (PTCs) in the cystic fibrosis transmembrane conductance regulator (CFTR) gene cause cystic fibrosis (CF). Several agents are known to suppress PTCs but are poorly efficacious or toxic.

Objectives: To determine whether there are clinically available agents that elicit translational readthrough and improve CFTR function sufficient to confer therapeutic benefit to patients with CF with PTCs.

Methods: Two independent screens, firefly luciferase and CFTR-mediated transepithelial chloride conductance assay, were performed on a library of 1,600 clinically approved compounds using fisher rat thyroid cells stably transfected with stop codons. Select agents were further evaluated using secondary screening assays including short circuit current analysis on primary cells from patients with CF. In addition, the effect of CFTR modulators (ivacaftor) was tested in combination with the most efficacious agents.

Measurements And Main Results: From the primary screen, 48 agents were selected as potentially active. Following confirmatory tests in the transepithelial chloride conductance assay and prioritizing agents based on favorable pharmacologic properties, eight agents were advanced for secondary screening. Ivacaftor significantly increased short circuit current following forskolin stimulation in cells treated with pyranoradine tetraphosphate, potassium p-aminobenzoate, and escin as compared with vehicle control. Escin, an herbal agent, consistently induced readthrough activity as demonstrated by enhanced CFTR expression and function in vitro.

Conclusions: Clinically approved drugs identified as potential readthrough agents, in combination with ivacaftor, may induce nonsense suppression to restore therapeutic levels of CFTR function. One or more agents may be suitable to advance to human testing.

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References

Santoro D, Postorino A, Costa S, Cristadoro S, Buemi M, Magazzu G . Renal amyloidosis in cystic fibrosis: role of colchicine therapy. Clin Kidney J. 2015; 7(1):81-2. PMC: 4389159. DOI: 10.1093/ckj/sft146. View

Raffetto J, Khalil R . Ca(2+)-dependent contraction by the saponoside escin in rat vena cava: implications in venotonic treatment of varicose veins. J Vasc Surg. 2011; 54(2):489-96. PMC: 3140579. DOI: 10.1016/j.jvs.2011.01.043. View

Kerem E, Konstan M, De Boeck K, Accurso F, Sermet-Gaudelus I, Wilschanski M . Ataluren for the treatment of nonsense-mutation cystic fibrosis: a randomised, double-blind, placebo-controlled phase 3 trial. Lancet Respir Med. 2014; 2(7):539-47. PMC: 4154311. DOI: 10.1016/S2213-2600(14)70100-6. View

Rowe S, Sloane P, Tang L, Backer K, Mazur M, Buckley-Lanier J . Suppression of CFTR premature termination codons and rescue of CFTR protein and function by the synthetic aminoglycoside NB54. J Mol Med (Berl). 2011; 89(11):1149-61. PMC: 3204584. DOI: 10.1007/s00109-011-0787-6. View

Wilschanski M, Yahav Y, Yaacov Y, Blau H, Bentur L, Rivlin J . Gentamicin-induced correction of CFTR function in patients with cystic fibrosis and CFTR stop mutations. N Engl J Med. 2003; 349(15):1433-41. DOI: 10.1056/NEJMoa022170. View

Sloane P, Rowe S . Cystic fibrosis transmembrane conductance regulator protein repair as a therapeutic strategy in cystic fibrosis. Curr Opin Pulm Med. 2010; 16(6):591-7. PMC: 3733473. DOI: 10.1097/MCP.0b013e32833f1d00. View

Xue X, Mutyam V, Tang L, Biswas S, Du M, Jackson L . Synthetic aminoglycosides efficiently suppress cystic fibrosis transmembrane conductance regulator nonsense mutations and are enhanced by ivacaftor. Am J Respir Cell Mol Biol. 2013; 50(4):805-16. PMC: 4068923. DOI: 10.1165/rcmb.2013-0282OC. View

Grentzmann G, Ingram J, Kelly P, GESTELAND R, Atkins J . A dual-luciferase reporter system for studying recoding signals. RNA. 1998; 4(4):479-86. PMC: 1369633. View

Nickless A, Jackson E, Marasa J, Nugent P, Mercer R, Piwnica-Worms D . Intracellular calcium regulates nonsense-mediated mRNA decay. Nat Med. 2014; 20(8):961-6. PMC: 4126864. DOI: 10.1038/nm.3620. View

10.

Van Goor F, Hadida S, Grootenhuis P, Burton B, Stack J, Straley K . Correction of the F508del-CFTR protein processing defect in vitro by the investigational drug VX-809. Proc Natl Acad Sci U S A. 2011; 108(46):18843-8. PMC: 3219147. DOI: 10.1073/pnas.1105787108. View

11.

Sermet-Gaudelus I, Renouil M, Fajac A, Bidou L, Parbaille B, Pierrot S . In vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study. BMC Med. 2007; 5:5. PMC: 1852113. DOI: 10.1186/1741-7015-5-5. View

12.

Zhang Y, Yu B, Sui Y, Gao X, Yang H, Ma T . Identification of resveratrol oligomers as inhibitors of cystic fibrosis transmembrane conductance regulator by high-throughput screening of natural products from chinese medicinal plants. PLoS One. 2014; 9(4):e94302. PMC: 3979805. DOI: 10.1371/journal.pone.0094302. View

13.

Sirtori C . Aescin: pharmacology, pharmacokinetics and therapeutic profile. Pharmacol Res. 2001; 44(3):183-93. DOI: 10.1006/phrs.2001.0847. View

14.

Sermet-Gaudelus I, Stoven V, Annereau J, Witko-Sarsat V, Reinert P, Guyot M . Interest of colchicine for the treatment of cystic fibrosis patients. Preliminary report. Mediators Inflamm. 2000; 8(1):13-5. PMC: 1781779. DOI: 10.1080/09629359990667. View

15.

Clancy J, Rowe S, Bebok Z, Aitken M, Gibson R, Zeitlin P . No detectable improvements in cystic fibrosis transmembrane conductance regulator by nasal aminoglycosides in patients with cystic fibrosis with stop mutations. Am J Respir Cell Mol Biol. 2007; 37(1):57-66. PMC: 1899350. DOI: 10.1165/rcmb.2006-0173OC. View

16.

Gonzalez-Hilarion S, Beghyn T, Jia J, Debreuck N, Berte G, Mamchaoui K . Rescue of nonsense mutations by amlexanox in human cells. Orphanet J Rare Dis. 2012; 7:58. PMC: 3562214. DOI: 10.1186/1750-1172-7-58. View

17.

Clancy J, Bebok Z, Ruiz F, King C, Jones J, Walker L . Evidence that systemic gentamicin suppresses premature stop mutations in patients with cystic fibrosis. Am J Respir Crit Care Med. 2001; 163(7):1683-92. DOI: 10.1164/ajrccm.163.7.2004001. View

18.

Pittler M, Ernst E . Horse chestnut seed extract for chronic venous insufficiency. Cochrane Database Syst Rev. 2004; (2):CD003230. DOI: 10.1002/14651858.CD003230.pub2. View

19.

Chen L, Yu B, Zhang Y, Gao X, Zhu L, Ma T . Bioactivity-guided fractionation of an antidiarrheal Chinese herb Rhodiola kirilowii (Regel) Maxim reveals (-)-epicatechin-3-gallate and (-)-epigallocatechin-3-gallate as inhibitors of cystic fibrosis transmembrane conductance regulator. PLoS One. 2015; 10(3):e0119122. PMC: 4352019. DOI: 10.1371/journal.pone.0119122. View

20.

Van Goor F, Hadida S, Grootenhuis P, Burton B, Cao D, Neuberger T . Rescue of CF airway epithelial cell function in vitro by a CFTR potentiator, VX-770. Proc Natl Acad Sci U S A. 2009; 106(44):18825-30. PMC: 2773991. DOI: 10.1073/pnas.0904709106. View