A small molecule that induces translational readthrough of CFTR nonsense mutations by eRF1 depletion

Jyoti Sharma; Ming Du; Eric Wong; Venkateshwar Mutyam; Yao Li; Jianguo Chen; Jamie Wangen; Kari Thrasher; Lianwu Fu; Ning Peng; Liping Tang; Kaimao Liu; Bini Mathew; Robert J Bostwick; Corinne E Augelli-Szafran; Hermann Bihler; Feng Liang; Jerome Mahiou; Josef Saltz; Andras Rab; Jeong Hong; Eric J Sorscher; Eric M Mendenhall; Candice J Coppola; Kim M Keeling; Rachel Green; Martin Mense; Mark J Suto; Steven M Rowe; David M Bedwell

doi:10.1038/s41467-021-24575-x

A small molecule that induces translational readthrough of CFTR nonsense mutations by eRF1 depletion

Nat Commun. 2021 Jul 16;12(1):4358. doi: 10.1038/s41467-021-24575-x.

Authors

Jyoti Sharma^#^{1

2

3}, Ming Du^#^{1

4}, Eric Wong⁵, Venkateshwar Mutyam^{1

2}, Yao Li^{1

2}, Jianguo Chen^{1

2}, Jamie Wangen⁶, Kari Thrasher^{1

4}, Lianwu Fu^{1

7}, Ning Peng^{1

2}, Liping Tang^{1

2}, Kaimao Liu^{1

4}, Bini Mathew⁸, Robert J Bostwick⁸, Corinne E Augelli-Szafran⁸, Hermann Bihler⁵, Feng Liang⁵, Jerome Mahiou⁵, Josef Saltz⁵, Andras Rab⁹, Jeong Hong⁹, Eric J Sorscher⁹, Eric M Mendenhall¹⁰, Candice J Coppola¹⁰, Kim M Keeling^{1

4}, Rachel Green⁶, Martin Mense⁵, Mark J Suto⁸, Steven M Rowe^{1

2

11}, David M Bedwell^{12

13}

Affiliations

¹ Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham (UAB), Birmingham, AL, USA.
² Department of Medicine, University of Alabama at Birmingham (UAB), Birmingham, AL, USA.
³ Department of Microbiology, University of Alabama at Birmingham (UAB), Birmingham, AL, USA.
⁴ Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham (UAB), Birmingham, AL, USA.
⁵ CFFT Lab, Cystic Fibrosis Foundation, Lexington, MA, USA.
⁶ Department of Molecular Biology and Genetics and Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁷ Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham (UAB), Birmingham, AL, USA.
⁸ Southern Research, Birmingham, AL, USA.
⁹ Department of Pediatrics, Emory University, Atlanta, Georgia.
¹⁰ Department of Biological Sciences, The University of Alabama in Huntsville, Huntsville, AL, USA.
¹¹ Department of Pediatrics, University of Alabama at Birmingham (UAB), Birmingham, AL, USA.
¹² Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham (UAB), Birmingham, AL, USA. dbedwell@uab.edu.
¹³ Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham (UAB), Birmingham, AL, USA. dbedwell@uab.edu.

^# Contributed equally.

Abstract

Premature termination codons (PTCs) prevent translation of a full-length protein and trigger nonsense-mediated mRNA decay (NMD). Nonsense suppression (also termed readthrough) therapy restores protein function by selectively suppressing translation termination at PTCs. Poor efficacy of current readthrough agents prompted us to search for better compounds. An NMD-sensitive NanoLuc readthrough reporter was used to screen 771,345 compounds. Among the 180 compounds identified with readthrough activity, SRI-37240 and its more potent derivative SRI-41315, induce a prolonged pause at stop codons and suppress PTCs associated with cystic fibrosis in immortalized and primary human bronchial epithelial cells, restoring CFTR expression and function. SRI-41315 suppresses PTCs by reducing the abundance of the termination factor eRF1. SRI-41315 also potentiates aminoglycoside-mediated readthrough, leading to synergistic increases in CFTR activity. Combining readthrough agents that target distinct components of the translation machinery is a promising treatment strategy for diseases caused by PTCs.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Aminoglycosides / metabolism
Codon, Nonsense / antagonists & inhibitors*
Codon, Nonsense / metabolism
Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
Cystic Fibrosis Transmembrane Conductance Regulator / metabolism
Epithelial Cells / drug effects*
Epithelial Cells / metabolism
Genes, Reporter
Gentamicins / pharmacology
HEK293 Cells
Humans
Microsomes, Liver / drug effects
Nonsense Mediated mRNA Decay*
Peptide Chain Termination, Translational / drug effects*
Peptide Termination Factors / genetics
Peptide Termination Factors / metabolism*
Proteasome Endopeptidase Complex / drug effects
Proteasome Endopeptidase Complex / metabolism
RNA Interference
Ribosomes / metabolism
Structure-Activity Relationship

Substances

Aminoglycosides
CFTR protein, human
Codon, Nonsense
ETF1 protein, human
Gentamicins
Peptide Termination Factors
Cystic Fibrosis Transmembrane Conductance Regulator
antibiotic G 418
Proteasome Endopeptidase Complex

Abstract

Publication types

MeSH terms

Substances

Grants and funding