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

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 com...

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Autores principales: Cócera-Ortega, Lucía, Wilders, Ronald, Kamps, Selina C., Fabrizi, Benedetta, Huber, Irit, van der Made, Ingeborg, van den Bout, Anouk, de Vries, Dylan K., Gepstein, Lior, Verkerk, Arie O., Pinto, Yigal M., Tijsen, Anke J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9000197/
https://www.ncbi.nlm.nih.gov/pubmed/35409410
http://dx.doi.org/10.3390/ijms23074053
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author Cócera-Ortega, Lucía
Wilders, Ronald
Kamps, Selina C.
Fabrizi, Benedetta
Huber, Irit
van der Made, Ingeborg
van den Bout, Anouk
de Vries, Dylan K.
Gepstein, Lior
Verkerk, Arie O.
Pinto, Yigal M.
Tijsen, Anke J.
author_facet Cócera-Ortega, Lucía
Wilders, Ronald
Kamps, Selina C.
Fabrizi, Benedetta
Huber, Irit
van der Made, Ingeborg
van den Bout, Anouk
de Vries, Dylan K.
Gepstein, Lior
Verkerk, Arie O.
Pinto, Yigal M.
Tijsen, Anke J.
author_sort Cócera-Ortega, Lucía
collection PubMed
description 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.
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spelling pubmed-90001972022-04-12 shRNAs Targeting a Common KCNQ1 Variant Could Alleviate Long-QT1 Disease Severity by Inhibiting a Mutant Allele Cócera-Ortega, Lucía Wilders, Ronald Kamps, Selina C. Fabrizi, Benedetta Huber, Irit van der Made, Ingeborg van den Bout, Anouk de Vries, Dylan K. Gepstein, Lior Verkerk, Arie O. Pinto, Yigal M. Tijsen, Anke J. Int J Mol Sci Article 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. MDPI 2022-04-06 /pmc/articles/PMC9000197/ /pubmed/35409410 http://dx.doi.org/10.3390/ijms23074053 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cócera-Ortega, Lucía
Wilders, Ronald
Kamps, Selina C.
Fabrizi, Benedetta
Huber, Irit
van der Made, Ingeborg
van den Bout, Anouk
de Vries, Dylan K.
Gepstein, Lior
Verkerk, Arie O.
Pinto, Yigal M.
Tijsen, Anke J.
shRNAs Targeting a Common KCNQ1 Variant Could Alleviate Long-QT1 Disease Severity by Inhibiting a Mutant Allele
title shRNAs Targeting a Common KCNQ1 Variant Could Alleviate Long-QT1 Disease Severity by Inhibiting a Mutant Allele
title_full shRNAs Targeting a Common KCNQ1 Variant Could Alleviate Long-QT1 Disease Severity by Inhibiting a Mutant Allele
title_fullStr shRNAs Targeting a Common KCNQ1 Variant Could Alleviate Long-QT1 Disease Severity by Inhibiting a Mutant Allele
title_full_unstemmed shRNAs Targeting a Common KCNQ1 Variant Could Alleviate Long-QT1 Disease Severity by Inhibiting a Mutant Allele
title_short shRNAs Targeting a Common KCNQ1 Variant Could Alleviate Long-QT1 Disease Severity by Inhibiting a Mutant Allele
title_sort shrnas targeting a common kcnq1 variant could alleviate long-qt1 disease severity by inhibiting a mutant allele
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9000197/
https://www.ncbi.nlm.nih.gov/pubmed/35409410
http://dx.doi.org/10.3390/ijms23074053
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