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shRNAs Targeting a Common KCNQ1 Variant Could Alleviate Long-QT1 Disease Severity by Inhibiting a Mutant Allele

Authors :
Lucía Cócera-Ortega
Ronald Wilders
Selina C. Kamps
Benedetta Fabrizi
Irit Huber
Ingeborg van der Made
Anouk van den Bout
Dylan K. de Vries
Lior Gepstein
Arie O. Verkerk
Yigal M. Pinto
Anke J. Tijsen
Medical Biology
ACS - Amsterdam Cardiovascular Sciences
Cardiology
ACS - Heart failure & arrhythmias
Graduate School
Source :
International Journal of Molecular Sciences; Volume 23; Issue 7; Pages: 4053, International journal of molecular sciences, 23(7):4053. Multidisciplinary Digital Publishing Institute (MDPI)
Publication Year :
2022
Publisher :
Multidisciplinary Digital Publishing Institute, 2022.

Abstract

Long-QT syndrome type 1 (LQT1) is caused by mutations in KCNQ1. Patients heterozygous for such a mutation co-assemble both mutant and wild-type KCNQ1-encoded subunits into tetrameric Kv7.1 potassium channels. Here, we investigated whether allele-specific inhibition of mutant KCNQ1 by targeting a common variant can shift the balance towards increased incorporation of the wild-type allele to alleviate the disease in human-induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs). We identified the single nucleotide polymorphisms (SNP) rs1057128 (G/A) in KCNQ1, with a heterozygosity of 27% in the European population. Next, we determined allele-specificity of short-hairpin RNAs (shRNAs) targeting either allele of this SNP in hiPSC-CMs that carry an LQT1 mutation. Our shRNAs downregulated 60% of the A allele and 40% of the G allele without affecting the non-targeted allele. Suppression of the mutant KCNQ1 allele by 60% decreased the occurrence of arrhythmic events in hiPSC-CMs measured by a voltage-sensitive reporter, while suppression of the wild-type allele increased the occurrence of arrhythmic events. Furthermore, computer simulations based on another LQT1 mutation revealed that 60% suppression of the mutant KCNQ1 allele shortens the prolonged action potential in an adult cardiomyocyte model. We conclude that allele-specific inhibition of a mutant KCNQ1 allele by targeting a common variant may alleviate the disease. This novel approach avoids the need to design shRNAs to target every single mutation and opens up the exciting possibility of treating multiple LQT1-causing mutations with only two shRNAs.

Details

Language :
English
ISSN :
14220067 and 16616596
Database :
OpenAIRE
Journal :
International Journal of Molecular Sciences; Volume 23; Issue 7; Pages: 4053
Accession number :
edsair.doi.dedup.....2ae9e01a9139de01455ee7a7b717fe33
Full Text :
https://doi.org/10.3390/ijms23074053