Cargando…
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...
Autores principales: | , , , , , , , , , , , |
---|---|
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 |
_version_ | 1784685375690637312 |
---|---|
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. |
format | Online Article Text |
id | pubmed-9000197 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT coceraortegalucia shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT wildersronald shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT kampsselinac shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT fabrizibenedetta shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT huberirit shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT vandermadeingeborg shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT vandenboutanouk shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT devriesdylank shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT gepsteinlior shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT verkerkarieo shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT pintoyigalm shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele AT tijsenankej shrnastargetingacommonkcnq1variantcouldalleviatelongqt1diseaseseveritybyinhibitingamutantallele |