Decreased mRNA and protein stability of W1282X limits response to modulator therapy

Journal of Cystic Fibrosis : Official Journal of the European Cystic Fibrosis Society
M A AksitNeeraj Sharma

Abstract

Cell-based studies have shown that W1282X generates a truncated protein that can be functionally augmented by modulators. However, modulator treatment of primary cells from individuals who carry two copies of W1282X generates no functional CFTR. To understand the lack of response to modulators, we investigated the effect of W1282X on CFTR RNA transcript levels. qRT-PCR and RNA-seq were performed on primary nasal epithelial (NE) cells of a previously studied individual who is homozygous for W1282X, her carrier parents and control individuals without nonsense variants in CFTR. CFTR RNA bearing W1282X in NE cells shows a steady-state level of 4.2 ± 0.9% of wild-type (WT) CFTR RNA in the mother and 12.4 ± 1.3% in the father. NMDI14, an inhibitor of nonsense-mediated mRNA decay (NMD), restored W1282X mRNA to almost 50% of WT levels in the parental NE cells. RNA-seq of the NE cells homozygous for W1282X showed that CFTR transcript level was reduced to 1.7% of WT (p-value: 4.6e-3). Negligible truncated CFTR protein was generated by Flp-In 293 cells stably expressing the W1282X EMG even though CFTR transcript was well above levels observed in the parents and proband. Finally, we demonstrated that NMD inhibition improved the stability a...Continue Reading

References

Mar 26, 2004·Nature Reviews. Molecular Cell Biology·Lynne E Maquat
Jun 2, 2007·American Journal of Respiratory Cell and Molecular Biology·Steven M RoweJohn P Clancy
Jul 13, 2007·European Journal of Human Genetics : EJHG·Liat LindeBatsheva Kerem
Aug 6, 2008·Nature Reviews. Genetics·Olaf Isken, Lynne E Maquat
Mar 6, 2009·Genome Biology·Ben LangmeadSteven L Salzberg
Mar 18, 2009·Bioinformatics·Cole TrapnellSteven L Salzberg
Aug 28, 2010·The Biochemical Journal·Madhuri BhuvanagiriAndreas E Kulozik
Oct 19, 2012·Journal of Bioinformatics and Computational Biology·Wentian Li
Dec 12, 2012·Nature Biotechnology·Cole TrapnellLior Pachter
Nov 19, 2013·Neuroscience and Biobehavioral Reviews·Lam Son NguyenJozef Gecz
Nov 21, 2013·American Journal of Respiratory Cell and Molecular Biology·Xiaojiao XueSteven M Rowe
Apr 8, 2014·EMBO Molecular Medicine·Yifat S OrenBatsheva Kerem
Apr 30, 2014·Annual Review of Genomics and Human Genetics·Kim M KeelingDavid M Bedwell
Jul 30, 2014·Nature Medicine·Andrew NicklessZhongsheng You
Oct 17, 2014·EMBO Molecular Medicine·Madhuri BhuvanagiriAndreas E Kulozik
Nov 19, 2014·Nature Reviews. Genetics·Garry R Cutting
Mar 4, 2015·Proceedings of the National Academy of Sciences of the United States of America·Bijoyita RoyAllan Jacobson
Oct 29, 2015·The New England Journal of Medicine·Abdul RehmanIbrahim A Janahi
Jan 17, 2016·Nucleic Acids Research·Nele HugJavier F Cáceres
Nov 1, 2016·American Journal of Respiratory and Critical Care Medicine·Venkateshwar MutyamSteven M Rowe
May 4, 2016·Pflügers Archiv : European journal of physiology·Franziska Ottens, Niels H Gehring
Oct 7, 2016·Journal of Cystic Fibrosis : Official Journal of the European Cystic Fibrosis Society·Venkateshwar MutyamSteven M Rowe
Nov 4, 2017·The New England Journal of Medicine·Steven M RoweJane C Davies
Nov 4, 2017·The New England Journal of Medicine·Jennifer L Taylor-CousarJ Stuart Elborn
May 29, 2018·American Journal of Human Genetics·Karen S RaraighGarry R Cutting
Aug 3, 2018·Nature·Daniel T MontoroJayaraj Rajagopal

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Citations

Jan 23, 2020·American Journal of Primatology·Jennifer L Taylor-CousarNeeraj Sharma
Apr 14, 2020·Pediatric Pulmonology·Jacelyn E Peabody LeverSteven M Rowe
Mar 11, 2020·Frontiers in Pharmacology·Miquéias Lopes-Pacheco
Oct 16, 2020·Scientific Reports·Rajdeep KaurSakthivel Murugan
May 1, 2021·International Journal of Molecular Sciences·Duncan E Keegan, John J Brewington
Feb 23, 2021·American Journal of Respiratory Cell and Molecular Biology·Venkateshwar MutyamSteven M Rowe
Jun 10, 2021·Journal of Cystic Fibrosis : Official Journal of the European Cystic Fibrosis Society·Lúcia SantosCarlos M Farinha
Jul 3, 2021·International Journal of Molecular Sciences·Christelle Bergeron, André M Cantin
Jun 4, 2020·Molecular Therapy. Methods & Clinical Development·Steven ErwoodEvgueni A Ivakine

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